Na-K-ATPase in the Enamel Organ: Localization and Possible Roles in Enamel Formation

1987 ◽  
Vol 1 (2) ◽  
pp. 267-275 ◽  
Author(s):  
P.R. Garant ◽  
T. Sasaki ◽  
P.E. Colflesh

Ouabain-sensitive, K-dependent p-nitrophenyl phosphatase (p-NPPase) activity was localized ultra-Ocytochemically in the lateral plasma membranes of secretory ameloblasts and the stratum intermedium and principally in the papillary layer cells of aldehyde-fixed rat incisor enamel organs by the one-step lead method. Daily intraperitoneal injection of ouabain (250 μg, 500 μg, and 1 mg/100 g body weight) for two weeks reduced p-NPPase activity in the enamel organ cells. However, the degree to which this activity was reduced appeared to vary among the experimental animals. Addition of ouabain to the cytochemical incubation medium completely inhibited p-NPPase activity in the tissues. Although long-term ouabain injection did not result in any morphological alterations of the enamel organ cells, it caused, in part, an appearance of electron-dense, homogeneous matrix-like substances (MS) in the extracellular spaces of the ameloblast layers at both the secretion and maturation stages. In addition, long-term ouabain injection appeared to have resulted in delayed maturation of enamel as measured by energy-dispersive x-ray analysis of Ca and P in surface enamel. These results suggest that Na-K-ATPase of enamel organ cells may participate in the net flow (removal) of organic matrix components and water from the enamel during the maturation stage of enamel formation. It is suggested that this flow is maintained by local osmotic gradients generated by Na-K-ATPase within the papillary layer.

1987 ◽  
Vol 1 (2) ◽  
pp. 213-226 ◽  
Author(s):  
T. Sasaki ◽  
D.E. Colflesh ◽  
P.R. Garant

Using aldehyde-fixed rat incisor enamel organ, we localized Ca-ATPase activity ultracytochemically in the plasma membranes, the mitochondrial inner membranes, and the Golgi membranes of secretory ameloblasts and the cells of stratum intermedium at the secretory stage and papillary layer cells at the maturation stage, but not in maturation ameloblasts. This Ca-ATPase activity was totally dependent on substrate ATP, the enzyme activator CaCl2, and also sensitive to the specific calmodulin blocker trifluoperazine (TFP) in the incubation media. Specific antigenic sites of endogenous calmodulin were demonstrated in polyribosomes, the nucleus, mitochondria, and the cytoplasmic matrix along the plasma membranes of secretory ameloblasts, by the protein A-immunogold technique using sheep antiserum against bovine testis calmodulin. All other enamel organ cells-such as stratum intermedium, papillary layer cells, and maturation ameloblasts-were also weakly immunoreactive. In control sections incubated with antiserum pre-absorbed with an excess of calmodulin and protein A-gold complex, only a few gold particles were observed to be randomly associated with the tissues. Daily intraperitoneal injection of TFP (1 and 5 mg per 100 g body weight) for one week resulted in prominent migration of mitochondria from the infranuclear to supranuclear regions of secretory ameloblasts but caused no other morphological alterations in the enamel organ cells. EDX analysis of ultrathin sections revealed significantly lower peaks of Ca and P in the forming enamel of TFP-injected rats than those in controls. However, little reduction in the Ca and P levels in the maturing enamel was observed in TFP-injected rats. When growing enamel surfaces were exposed with NaOCl and examined with SEM, a remarkable defect in the enamel matrix was observed in the forming enamel but not in the maturing enamel. These results suggest that early enamel mineralization is dependent upon an intact calmodulin-regulated Ca-transporting ATPase in secretory ameloblasts and that enamel maturation is controlled by different mechanism(s).


2008 ◽  
Vol 389 (6) ◽  
Author(s):  
Yuhe Lu ◽  
Petros Papagerakis ◽  
Yasuo Yamakoshi ◽  
Jan C.-C. Hu ◽  
John D. Bartlett ◽  
...  

Abstract Two proteases are secreted into the enamel matrix of developing teeth. The early protease is enamelysin (MMP-20). The late protease is kallikrein 4 (KLK4). Mutations in MMP20 and KLK4 both cause autosomal recessive amelogenesis imperfecta, a condition featuring soft, porous enamel containing residual protein. MMP-20 is secreted along with enamel proteins by secretory-stage ameloblasts. Enamel protein-cleavage products accumulate in the space between the crystal ribbons, helping to support them. MMP-20 steadily cleaves accumulated enamel proteins, so their concentration decreases with depth. KLK4 is secreted by transition- and maturation-stage ameloblasts. KLK4 aggressively degrades the retained organic matrix following the termination of enamel protein secretion. The principle functions of MMP-20 and KLK4 in dental enamel formation are to facilitate the orderly replacement of organic matrix with mineral, generating an enamel layer that is harder, less porous, and unstained by retained enamel proteins.


1989 ◽  
Vol 37 (5) ◽  
pp. 581-587 ◽  
Author(s):  
D M Lyaruu ◽  
W J Lenglet ◽  
J H Wöltgens ◽  
A L Bronckers

A micro-PIGE (Proton-Induced gamma-ray Emission) technique based on the delayed 5/2+----1/2+ nuclear transition of fluorine (E gamma = 197 keV, t1/2 = 87 ns) emitted after 19F(p,p', gamma)19F reaction was used to detect and study the distribution of fluorine in the developing enamel organ during pre-eruptive stages, i.e., the transitional to early maturation stages of enamel formation in neonatal hamsters administered a single IP dose of sodium fluoride (20 mg NaF/kg body weight). The aforementioned nuclear reaction is unique for fluorine, and therefore detection of gamma-rays emanating from this reaction in a biological specimen implies a positive identification of fluorine at that particular site. Calcium and phosphorus X-rays were also recorded and used as parameters for assessment of the relationship between the degree of mineralization and fluoride incorporation into the enamel organ. The highest fluorine concentration in the enamel organ was recorded in the dentin near the dentin-enamel junction (DEJ). In the enamel, the highest concentration of fluorine was found to be associated with the more mature areas of the enamel near the DEJ, but gradually decreased in the direction of the enamel surface. Fluorine was not detected in the control germs. These results suggest that administration of fluoride in high doses during the pre-eruptive stages of enamel formation leads to incorporation of the ion into the forming dentin and enamel mineral, and that the enamel matrix does not seem to bind fluoride avidly.


1987 ◽  
Vol 66 (8) ◽  
pp. 1360-1363 ◽  
Author(s):  
J.W. Bawden ◽  
T.G. Deaton ◽  
M.A. Crenshaw

This study investigated the diffusion of fluoride through the enamel organ in vitro. The rat molar explants used were entirely in the secretory stage or predominantly in the maturation stage of enamel formation. The removal of the enamel organ or metabolic inhibition with iodoacetate caused significant increases in enamel fluoride uptake at both stages of enamel formation. Inhibition with dinitrophenol caused a significant increase only in the maturation phase. Uptake of fluoride in enamel was related to the fluoride concentration in the medium, except in the maturation stage explants, where increasing the medium fluoride concentration from 0.05 ppm to 0.08 ppm did not significantly increase fluoride uptake at any of the three observation times. The findings indicate that the enamel organ exists as a diffusion-limiting membrane to the movement of fluoride from the extracellular fluid compartment to the developing enamel.


1999 ◽  
Vol 10 (4) ◽  
pp. 425-441 ◽  
Author(s):  
J.D. Bartlett ◽  
J.P. Simmer

For almost three decades, proteinases have been known to reside within developing dental enamel. However, identification and characterization of these proteinases have been slow and difficult, because they are present in very small quantities and they are difficult to purify directly from the mineralizing enamel. Enamel matrix proteins such as amelogenin, ameloblastin, and enamelin are cleaved by proteinases soon after they are secreted, and their cleavage products accumulate in the deeper, more mature enamel layers, while the full-length proteins are observed only at the surface. These results suggest that proteinases are necessary for "activating" enamel proteins so the parent proteins and their cleavage products may perform different functions. A novel matrix metalloproteinase named enamelysin (MMP-20) was recently cloned from tooth tissues and was later shown to localize primarily within the most recently formed enamel. Furthermore, recombinant porcine enamelysin was demonstrated to cleave recombinant porcine amelogenin at virtually all of the sites that have previously been described in vivo. Therefore, enamelysin is at least one enzyme that may be important during early enamel development. As enamel development progresses to the later stages, a profound decrease in the enamel protein content is observed. Proteinases have traditionally been assumed to degrade the organic matrix prior to its removal from the enamel. Recently, a novel serine proteinase named enamel matrix serine proteinase-1 (EMSP1) was cloned from enamel organ epithelia. EMSP1 localizes primarily to the early maturation stage enamel and may, therefore, be involved in the degradation of proteins prior to their removal from the maturing enamel. Other, as yet unidentified, proteinases and proteinase inhibitors are almost certainly present within the forming enamel and await discovery.


1987 ◽  
Vol 1 (2) ◽  
pp. 176-180 ◽  
Author(s):  
P.K. Denbesten ◽  
M.A. Crenshaw

Exposure to chronic high levels of fluoride results in the formation of fluorosed enamel. Although enamel may be more susceptible to fluorotic effects at certain stages of development, fluoride at sufficiently high levels may affect enamel at all stages of formation. Careful study of the changes in enamel caused by chronic fluoride ingestion is needed to understand more fully the mechanisms involved in the formation of fluorotic enamel. This paper discusses the various studies we have completed to define the changes, in developing enamel of the rat incisor, caused by long-term ingestion of fluoride in drinking water. Fluoride has been found to inhibit secretion of enamel proteins. Changes in the maturation stage of enamel formation include the retention of amelogenin proteins during early maturation. The various mechanisms which have been investigated in the formation of fluorosed enamel include a direct effect of fluoride on the enamel organ, and specific interactions of fluoride with the extracellular enamel matrix. Although the same amount of protease appears to be secreted in fluorosed and control enamel, a delay in the digestion of amelogenin protein occurs. This suggests that fluoride may directly or indirectly inhibit the protease present in fluorosed enamel to slow the proteolysis of amelogenins.


2005 ◽  
Vol 44 (03) ◽  
pp. 107-117
Author(s):  
R. G. Meyer ◽  
W. Herr ◽  
A. Helisch ◽  
P. Bartenstein ◽  
I. Buchmann

SummaryThe prognosis of patients with acute myeloid leukaemia (AML) has improved considerably by introduction of aggressive consolidation chemotherapy and haematopoietic stem cell transplantation (SCT). Nevertheless, only 20-30% of patients with AML achieve long-term diseasefree survival after SCT. The most common cause of treatment failure is relapse. Additionally, mortality rates are significantly increased by therapy-related causes such as toxicity of chemotherapy and complications of SCT. Including radioimmunotherapies in the treatment of AML and myelodyplastic syndrome (MDS) allows for the achievement of a pronounced antileukaemic effect for the reduction of relapse rates on the one hand. On the other hand, no increase of acute toxicity and later complications should be induced. These effects are important for the primary reduction of tumour cells as well as for the myeloablative conditioning before SCT.This paper provides a systematic and critical review of the currently used radionuclides and immunoconjugates for the treatment of AML and MDS and summarizes the literature on primary tumour cell reductive radioimmunotherapies on the one hand and conditioning radioimmunotherapies before SCT on the other hand.


2018 ◽  
pp. 49-68 ◽  
Author(s):  
M. E. Mamonov

Our analysis documents that the existence of hidden “holes” in the capital of not yet failed banks - while creating intertemporal pressure on the actual level of capital - leads to changing of maturity of loans supplied rather than to contracting of their volume. Long-term loans decrease, whereas short-term loans rise - and, what is most remarkably, by approximately the same amounts. Standardly, the higher the maturity of loans the higher the credit risk and, thus, the more loan loss reserves (LLP) banks are forced to create, increasing the pressure on capital. Banks that already hide “holes” in the capital, but have not yet faced with license withdrawal, must possess strong incentives to shorten the maturity of supplied loans. On the one hand, it raises the turnovers of LLP and facilitates the flexibility of capital management; on the other hand, it allows increasing the speed of shifting of attracted deposits to loans to related parties in domestic or foreign jurisdictions. This enlarges the potential size of ex post revealed “hole” in the capital and, therefore, allows us to assume that not every loan might be viewed as a good for the economy: excessive short-term and insufficient long-term loans can produce the source for future losses.


2018 ◽  
Vol 59 (1) ◽  
pp. 65-79
Author(s):  
Katarzyna Nikorowicz-Zatorska

Abstract The present paper focuses on spatial management regulations in order to carry out investment in the field of airport facilities. The construction, upgrades, and maintenance of airports falls within the area of responsibility of local authorities. This task poses a great challenge in terms of organisation and finances. On the one hand, an active airport is a municipal landmark and drives local economic, social and cultural development, and on the other, the scale of investment often exceeds the capabilities of local authorities. The immediate environment of the airport determines its final use and prosperity. The objective of the paper is to review legislation that affects airports and the surrounding communities. The process of urban planning in Lodz and surrounding areas will be presented as a background to the problem of land use management in the vicinity of the airport. This paper seeks to address the following questions: if and how airports have affected urban planning in Lodz, does the land use around the airport prevent the development of Lodz Airport, and how has the situation changed over the time? It can be assumed that as a result of lack of experience, land resources and size of investments on one hand and legislative dissonance and peculiar practices on the other, aviation infrastructure in Lodz is designed to meet temporary needs and is characterised by achieving short-term goals. Cyclical problems are solved in an intermittent manner and involve all the municipal resources, so there’s little left to secure long-term investments.


2019 ◽  
Vol 10 (1) ◽  
pp. 21-28
Author(s):  
Aniela Bălăcescu ◽  
Radu Șerban Zaharia

Abstract Tourist services represent a category of services in which the inseparability of production and consumption, the inability to be storable, the immateriality, and last but not least non-durability, induces in tourism management a number of peculiarities and difficulties. Under these circumstances the development of medium-term strategies involves long-term studies regarding on the one hand the developments and characteristics of the demand, and on the other hand the tourist potential analysis at regional and local level. Although in the past 20 years there has been tremendous growth of on-line booking made by household users, the tour operators agencies as well as those with sales activity continue to offer the specific services for a large number of tourists, that number, in the case of domestic tourism, increased by 1.6 times in case of the tour operators and by 4.44 times in case of the agencies with sales activity. At the same time, there have been changes in the preferences of tourists regarding their holiday destinations in Romania. Started on these considerations, paper based on a logistic model, examines the evolution of the probabilities and scores corresponding to the way the Romanian tourists spend their holidays on the types of tourism agencies, actions and tourist areas in Romania.


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