Gender difference in unstimulated whole saliva flow rate and salivary gland sizes.

OBJECTIVE: A gender difference in the unstimulated whole saliva flow rate (UWSFR) may be due to a difference in the sizes of the salivary glands. In this study, we investigated the relationships among the UWSFR, gland sizes and body sizes of healthy young adult males and females. DESIGN: Unstimulated whole saliva was collected for 5 min by the spitting method in 50 healthy young adults, and the flow rate of the saliva was measured. Heights and weights were measured, and body mass indices (BMI) were calculated. The sizes of the salivary glands were measured by use of a magnetic resonance imaging technique. RESULTS: Parotid and submandibular gland sizes and flow rates in females were significantly smaller than those in males, as were also the weights, heights and BMI. In both males and females, there were significant positive correlations between gland sizes and the flow rates, weights and BMI. The variations of the flow rates were reduced by standardizing them with gland sizes, weights and BMI. CONCLUSIONS: These results suggest that the lower UWSFR in females as compared with males is due to the smaller gland sizes due to the smaller body sizes.

Oscillation of ADP-ribosyl cyclase activity during the cell cycle and function of cyclic ADP-ribose in a unicellular organism, Euglena gracilis.

In Euglena gracilis, the activity of ADP-ribosyl cyclase, which produces cyclic ADP-ribose, oscillated during the cell cycle in a synchronous culture induced by a light-dark cycle, and a marked increase in the activity was observed in the G2 phase. Similarly, the ADP-ribosyl cyclase activity rose extremely immediately before cell division started, when synchronous cell division was induced by adding cobalamin (which is an essential growth factor and participates in DNA synthesis in this organism) to its deficient culture. Further, cADPR in these cells showed a maximum level immediately before cell division started. A dose-dependent Ca2+ release was observed when microsomes were incubated with cADPR.

Purification and characterization of arginine:mono-ADP-ribosylhydrolase from Euglena gracilis Z.

Arginine:mono-ADP-ribosylhydrolase was purified from a protozoan, Euglena gracilis Z, using [32P]mono-ADP-ribosylated actin as a substrate. The enzyme showed molecular mass of 33 kDa both in SDS PAGE and gel filtration, indicating it to be a monomeric protein. It was strongly inhibited by ADP and ADP-ribose and activated by Mg2+, DTT, and 2-mercaptoethanol. These results suggest that it recognizes the ADP-ribose moiety of the modified protein. Since the enzyme activity increased in S phase and late G0 phase in a synchronous dividing culture, the enzyme may function in the regulation of the cell cycle.

Purification and characterization of ADP-ribosyl cyclase from Euglena gracilis.

ADP-ribosyl cyclase, which catalyzes the conversion from NAD+ to cyclic adenosine diphosphoribose (cADPR), is proposed to participate in cell cycle regulation in Euglena gracilis. This enzyme, which was found as a membrane-bound protein, was purified almost the homogeneity after solubilization with deoxycholate, and found to be a monomeric protein with a molecular mass of 40 kDa. Its Km value for NAD+ was estimated to be 0.4 mM, and cADPR, a product of the enzyme, inhibited the enzyme competitively with respect to NAD+ whereas another product, nicotinamide, showed noncompetitive (mixed-type) inhibition. In contrast to mammalian CD38 and BST-1, Euglena ADP-ribosyl cyclase lacked cADPR hydrolase activity.

Inositol 1,4,5-trisphosphate and cyclic ADP-ribose mobilize Ca2+ in a protist, Euglena gracilis.

Inositol 1,4,5-trisphosphate (InsP3) and cyclic ADP-ribose (cADPR) released Ca2+ from microsome fraction prepared from Euglena gracilis in dose-dependent manners. Caffeine, which also induced Ca2+ release from the microsomes, caused desensitization of the Ca2+ response to cADPR, although the Ca2+ response to InsP3 was not affected by caffeine. Further, ruthenium red inhibited the Ca2+ release induced by cADPR, but not by InsP3. These results suggest that cADPR functions as an endogenous messenger to activate a caffeine-sensitive, Ca(2+)-release mechanism, whereas InsP3 induces Ca2+ release by a distinct mechanism in E. gracilis.

[Hemostatic effect of an endoscopic local injection of hypertonic saline-epinephrine (HS-E) solution on upper gastrointestinal bleeding--comparative study of this effect with and without visible vessel detection].

One hundred and ninety seven patients with upper gastrointestinal bleeding were each administered and endoscopic local injection of hypertonic saline epinephrine (HS-E) solution at our hospital over a 5-year period, from the Spring of 1981 to March of 1986. Peptic ulcers were found in 162 of the patient. Vessels could be visibly detected in the 148 (91.4%)-A group, but not in the 14 (8.6%)-B group. A comparison of the number of endoscopic local injections to each of these groups indicated no significant differences in types of bleeding, systemic complications, or sites of bleeding. The excellent hemostatic effect of HS-E is due to the fact that local injection of its solution can be repeated in large quantities for various types of bleeding.

Mineral trioxide aggregate inhibits osteoclastic bone resorption.

Mineral trioxide aggregate (MTA), a commonly used endodontic repair material, is useful for both basic and clinical research, and the effect of MTA on osteoblast differentiation has been well-defined. However, the effects of MTA on osteoclastic bone resorption are not fully understood. Hence, the aim of this study is to examine the effect of MTA solution in the regulation of osteoclast bone-resorbing activity using osteoclasts formed in co-cultures of primary osteoblasts and bone marrow cells. MTA solution dose-dependently reduced the total area of pits formed by osteoclasts. The reduction of resorption induced by 20% MTA treatment was due to inhibition of osteoclastic bone-resorbing activity and had no effect on osteoclast number. A 20% MTA solution disrupted actin ring formation, a marker of osteoclastic bone resorption, by reducing phosphorylation and kinase activity of c-Src, and mRNA expressions of cathepsin K and mmp-9. A high concentration of MTA solution (50%) induced apoptosis of osteoclasts by increasing the expression of Bim, a member of the BH3-only (Bcl-2 homology) family of pro-apoptotic proteins. Taken together, our results suggest that MTA is a useful retrofilling material for several clinical situations because it both stimulates osteoblast differentiation and inhibits bone resorption.

ADP-Ribosyl cyclase in rat salivary glands.

Both the Ca(2+)-releasing mechanism induced by cyclic ADP-ribose (cADPR) and the ADP-ribosyl cyclase (ADPRC) activity that converts NAD(+) to cADPR were observed in a variety of cell types. We studied the ADPRC activity in rat major salivary glands that include parotid gland (PG), submandiblar gland (SMG), and sublingual gland (SLG). The enzyme activity responsible for cADPR synthesis was detected by spectrofluorometric assay using NGD(+) as a substrate. The enzyme activities in SLG, SMG, and PG were about 400, 30, and 40 nmol/min/g tissue, respectively, in 5-week-old rats. The highest value was observed in SLG and this value was higher than those in other tissues; e.g., spleen (200 nmol/min/g tissue). The enzyme activity in SLG increased gradually after birth and showed a maximum value at 3 weeks. On the other hand, the enzyme activities almost did not change in both PG and SMG between 0 and 9 weeks. In spite of the high ADPRC activity in SLG, we could not detect the cADPR-induced Ca(2+)-release from SLG microsomes. These results suggest that the ADPRC in SLG does not function through Ca(2+)-release observed in various tissues.

A new type of allantoinase in amphibian liver.

Allantoinase and allantoicase are known to form a complex in amphibian liver. In this study, a new type of allantoinase that did not form a complex with allantoicase was found in the amphibian liver. Purified enzyme had a molecular mass of about 44 kDa both in SDS-PAGE and gel-filtrations. The enzyme cross-reacted with anti-sardine allantoinase polyclonal antibody, and it weakly cross-reacted with anti-bullfrog allantoinase polyclonal antibody.