ABO-incompatible pediatric kidney transplantation without antibody removal.

Background: Because of the severe shortage of suitable deceased donors, ABO-incompatible living donor kidney transplantation (ABOi LDKT) is performed even in pediatric recipients in Japan. We performed pediatric ABOi LDKT using rituximab without anti-A/B antibody removal.

Methods: Thirteen pediatric recipients (mean age 7.

Prolactin enhances CCAAT enhancer-binding protein-beta (C/EBP beta) and peroxisome proliferator-activated receptor gamma (PPAR gamma) messenger RNA expression and stimulates adipogenic conversion of NIH-3T3 cells.

Extracellular stimuli trigger adipocyte differentiation by inducing the complex cascades of transcription. Transcription factors CCAAT enhancer-binding proteins (C/EBPs) and peroxisome proliferator-activated receptor gamma (PPARgamma) play crucial roles in this process. Although ectopic expression of these factors in NIH-3T3 cells, a multipotential mesenchymal stem cell line, results in adipogenic conversion, little is known as to hormonal factors that regulate adipogenesis in these cells.

Escherichia coli prlC gene encodes a trypsin-like proteinase regulating the cell cycle.

Proteinase In has previously been described as displaying a trypsin-like proteinase activity that momentarily appears immediately before DNA synthesis in the cell cycle of Escherichia coli synchronized by phosphate starvation and which is closely related to the initiation of DNA replication [Kato, M., Irisawa, T., Ohtani, M.

Effects of amidinopiperidine-4-carboxylic acid 4-tert-butylphenyl ester, a specific trypsin inhibitor, on the growth of HL-60 cells.

Previous results showed that the synthetic compound amidinopiperidine-4-carboxylic acid 4-tert-butylphenyl ester (APCA-OPhBut), a trypsin inhibitor, could specifically inhibit the activity of proteinase In and lead to growth arrest of Hela cells in early S phase. In this study, APCA-OPhBut exhibited inhibitory effects on the growth of HL-60 cells. Apoptotic cells were observed when the cells were cultured with APCA-OPhBut above 50 microM.

Partial purification of a trypsin-like proteinase in platelets.

Among various fluorogenic substrates for trypsin-like proteinases, tert-butyloxycarbonyl-L-valyl-L-prolyl-L-arginine 4-methylcoumarin-7-amide was strongly hydrolyzed by a crude extract of rabbit platelets. The proteinase was partially purified (92-fold) from rabbit platelets by successive chromatographic separations on phenyl-Sepharose CL-4B, L-arginine-Sepharose 4B and Sephadex G-200 columns. Its molecular mass was found to be greater than 200 kDa by analytical gel filtration and its optimal pH was approximately 9.

Inhibition of trypsin, plasmin, thrombin and kallikrein by various esters of guanidino- and amidino-acids.

Inhibitory effects of various phenolic esters of trans-4-guanidinomethylcyclohexanecarboxylic acid, amidino-piperidine-4-alkanoic acids or trans-4-amidinocyclohexane-4-alkanoic acids on trypsin, thrombin, plasmin and pancreatic kallikrein were examined. Their inhibitory effects were strongly affected by the acid portion and phenolic group constituting the esters. The effects of the acidic portion and phenolic group on the inhibitory effect varied with each protease; they were most effective on thrombin and plasmin and least effective on kallikrein.

Properties of three proteinases functioning at G1, S and G2 phases in HeLa cells and their inhibition by guanidino- and amidino-acid esters.

HeLa cells were synchronized by double thymidine-block and allowed to grow after removal of thymidine. Three proteinases, tryptase 17:17, proteinase In and late G2 proteinase, were prepared from the HeLa cells harvested at the time when each proteinase appeared in the cell cycle of the cells. All of them were suggested to be trypsin-like serine proteinases, because they hydrolyzed trypsin-specific fluorogenic substrates and their activities were inhibited by benzamidine, soybean trypsin inhibitor, leupeptin, tosyl-L-lysine chloromethan (TLCK) and diisopropylfluorophosphate (DEP).

Antibacterial effects of esters of guanidino- and amidino-acids trypsin inhibitors.

Inhibitory effects of various esters of trans-4-guanidinomethylcyclohexanecarboxylic acid and the 4-tert- butylphenyl esters of amidinopiperidine-4-alkanoic, trans-4- amidinocyclohexanealkanoic, trans-4-guanidinoethylcyclohexanecarboxylic and trans-4-guanidinocyclohexanealkanoic acids, all trypsin inhibitors, on the growth of E. coli, B. subtilis, S.

Effects of various esters of trans-4-guanidinomethylcyclohexanecarboxylic acid, trypsin inhibitors, on the growth of Bacillus subtilis.

Various aromatic esters of trans-4-guanidinomethylcyclohexanecarboxylic acid (GMCHA), trypsin inhibitors, strongly inhibited the growth of Bacillus subtilis 558 and their effects were markedly affected by the species of substitution on the phenyl nucleus of the GMCHA phenyl esters. 4-tert-Butylphenyl ester of GMCHA (GMCHA-OPhtBu), a representative of various GMCHA esters, dose-dependently inhibited the growth of B. subtilis and DNA, RNA and protein syntheses in the cells.

trans-4-amidinocyclohexanecarboxylic acid 4-tert-butylphenyl ester, a trypsin inhibitor, blocks entry of HeLa cells from G2 phase into mitosis.

Release of HeLa cells arrested at the G1/S boundary by double-thymidine block immediately caused uptake of [3H]thymidine into DNA. The duration of the cell cycle time was 23 h and definite changes in cell density were observed between 12 h and 13 h and also between 35 h and 36 h after removal of thymidine. Addition of trans-4-amidino-cyclohexanecarboxylic acid 4-tert-butylphenyl ester (ACHCA-OPhBut), immediately after removal of the arrest had no effect on DNA synthesis, although it dose-dependently suppressed the first mitosis and the next round of DNA synthesis.

Effect of trans-4-guanidinomethylcyclohexanecarboxylic acid 4-tert-butylphenyl ester, a trypsin inhibitor, on the growth of various strains of Escherichia coli.

Similarly to Escherichia coli K-12 IAM 1264, trans-4- guanidinomethylcyclohexanecarboxylic acid 4-tert-butylphenyl ester (GMCHA-OPhtBu), a trypsin inhibitor, had a strong inhibitory effect on the growth of various strains of E. coli K-12, such as W 3350, AB 1157, JM 103, W 3110, C 600r-m- and C 600r+m+, preceded by suppressive effects of GMCHA-OPhtBu on DNA synthesis in these strains, although the inhibitory effects varied from strain to strain. These results suggested the possible involvement of a trypsin-like proteinase in DNA synthesis.

Amidinopiperidine-4-carboxylic acid 4-tert-butylphenyl ester, a trypsin inhibitor, suppresses the onset of DNA synthesis in HeLa cells synchronized by a double-thymidine block.

Release of HeLa cells arrested at the G1/S boundary by double-thymidine block caused abrupt uptake of [methyl-3H]thymidine into DNA after 5 min, and two sharp high activity peaks, peak I and peak II, were observed 8 and 23 min after removal of the thymidine block and this was followed by a gradual uptake of [3H]thymidine. The duration of the cell cycle was 23 h, and definite changes in cell density were observed between 12 and 13 h and also between 35 and 36 h after removal of the thymidine. Addition of amidinopiperidine-4-carboxylic acid 4-tert-butylphenyl ester (APCA-OPh'Bu), a trypsin inhibitor, immediately after removal of the arrest strongly suppressed DNA synthesis and mitosis.

A trypsin inhibitor trans-4-guanidinomethylcyclohexanecarboxylic acid 4-tert-butylphenyl ester suppresses the onset of DNA synthesis in Escherichia coli cells synchronized by phosphate starvation.

trans-4-Guanidinomethylcyclohexanecarboxylic acid 4-tert-butylphenyl ester (GMCHA-OPh'Bu), a trypsin inhibitor, dose-dependently inhibited the growth of Escherichia coli K-12 IAM1264. Growth inhibition was preceded by dose- and time-dependent inhibition of DNA synthesis. These results strongly suggested participation of a trypsin-like proteinase in DNA synthesis.

Effects of synthetic trypsin inhibitors on the cell cycle of synchronized HeLa cells.

HeLa cells were synchronized by a double-thymidine block. After removal of thymidine, the cells immediately caused the uptake of [3H]thymidine into DNA and reached a half-maximum. The duration of the cell cycle was 23 h, and definite changes in cell density were observed between 12 and 13 h and between 35 and 36 h after removal of thymidine.

Effects of synthetic trypsin inhibitors on the growth of Escherichia coli.

The inhibitory effects of various aromatic esters of trans-4-guanidinomethylcyclohexanecarboxylic acid (GMCHA), potent trypsin inhibitors, on the growth of Escherichia coli were examined and the effects were compared with those of well known synthetic trypsin inhibitors. Various GMCHA esters strongly inhibited the growth of E. coli and their effects were markedly affected by the species and position of substitution on the phenyl nucleus of the GMCHA phenyl esters.

Purification and characterization of proteinase In, a trypsin-like proteinase, in Escherichia coli.

We previously found a trypsin-like proteinase which momentarily appears immediately before DNA synthesis in the cell cycle of Escherichia coli synchronized by phosphate starvation and which is closely related to the initiation of DNA replication (Kato, M., Irisawa, T., Morimoto, Y.

Inhibition of gastric H+, K(+)-ATPase and acid secretion by ellagic acid.

The effects of ellagic acid on gastric H+, K(+)-ATPase, acid secretion, and the occurrence of gastric ulcers were studied. Ellagic acid inhibited hog gastric H+, K(+)-ATPase activity with a 50% inhibition at 2.1 x 10(-6)M; kinetic studies showed that the inhibition of H+, K(+)-ATPase by ellagic acid is competitive with respect to ATP and is noncompetitive with respect to K+.

Inhibition of phospholipid methylation by an anti-allergic agent, NCO-650, during histamine release.

Antigen, anti-IgE and concanavalin A (Con A) induced an increase in both the incorporation of the 3H-methyl moiety into phospholipids and histamine release. Maximal incorporation of the 3H-methyl moiety into the lipid fraction of the cells was observed within 15 sec and 1 min after being challenged with antigen (100 micrograms/mL) and anti-IgE (200 micrograms/mL) respectively. However, the methylated phospholipid decreased rapidly.

A trypsin-like proteinase appearing at 17 h and 17 min in the cell cycle time of HeLa cells correlates with the onset of DNA synthesis.

A trypsin-like proteinase appearing sharply at 17 h 17 min (17:17) in the cell cycle time of the synchronized and growing HeLa cells correlates with the onset of DNA synthesis, and the inhibition of the proteinase activity by trans-4-guanidinomethylcyclohexanecarboxylic acid 4-tert-butylphenyl ester (GMCHA-OPhBut) results in a 3 h retardation of the onset of the DNA synthesis. The proteinase activity is cell density dependent and completely inhibited by 100 microM GMCHA-OPhBut. The proteinase was named HeLa tryptase 17:17.

Fluctuation of trypsin-like proteinase activity in the cell cycle of synchronized and growing HeLa cells, and the effect of GMCHA-OPhBut.

HeLa cells synchronized by double-thymidine block were grown in Eagle's minimum essential medium supplemented with 10% calf serum, and the fluctuation of trypsin-like protease activity in the cell cycle was examined. Seven distinct activity peaks were observed in one cell cycle at a cell density of 2%: two peaks in S phase, one peak at the S/G2 boundary, one peak in early M phase and one at the M/G1 boundary, and two peaks in G1 phase. HeLa cells synchronized by a mitotic detachment technique also showed similar results at cell density of 4.

Inhibitory effects of GMCHA-OPhBut on phospholipid methylation and histamine release in mast cells activated by concanavalin A, anti-IgE, and antigen.

[3H]Methyl group incorporation and histamine secretion in rat mast cells induced by anti-IgE and con A were strongly inhibited by trans-4-guanidinomethylcyclohexanecarboxylic acid 4-tert-butylphenyl ester (GMCHA-OPhBut), a strong and specific inhibitor for pH 7 tryptase (Muramatsu et al. (1988) Biol. Chem.

Inhibition of cAMP increase by an anti-allergic agent, NCO-650, during histamine release.

Antigen and concanavalin A (Con A) induced an increase in cAMP and histamine release from rat peritoneal mast cells. In a dose-dependent manner, the compound, NCO-650, significantly inhibited both the initial and secondary increases in cAMP stimulated by antigen, anti-IgE and Con A in rat peritoneal mast cells. IC50 values of NCO-650 for cAMP increase stimulated by antigen, anti-IgE and Con A were 3.

Role of calcium in histamine release from rat mast cells activated by various secretagogues; intracellular calcium mobilization correlates with histamine release.

Anti-IgE, con A or antigen caused an increase in the intracellular calcium concentration, [Ca2+]i, of mast cells. The increase occurred in two stages: a rapid initial rise caused by Ca-mobilization from intracellular Ca-stores and a second sustained rise caused by an influx of extracellular calcium (White, J.R.

Role of cyclic AMP during histamine release. Histamine release is not directly related to increase in cyclic AMP levels in rat mast cells activated by concanavalin A, anti-IgE, antigen, prostaglandin D2 and isoproterenol.

Activation of mast cells by bridging of IgE-receptors or concanavalin A (Con A) results in a rapid initial rise and fall in cyclic AMP (cAMP) levels followed by a second rise in cAMP levels and histamine release (Sullivan, T. et al. (1976) J.

Inhibitory effect of anti-allergic agent NCO-650 on histamine release induced by various secretagogues.

Histamine release from rat peritoneal mast cells induced by antigen and anti-IgE was essentially complete within 2 min and 3 min, respectively, but that due to Concanavalin A (Con A) was complete only within 9 min. An anti-allergic agent NCO-650 [trans-4-Guanidinomethylcyclohexanecarboxylic acid p-tert-butylphenyl ester hydrochloride], which is a strong inhibitor of trypsin, dose-dependently inhibited anti-IgE-induced histamine release from rat peritoneal mast cells. Moreover, the rate and extent of histamine release from rat peritoneal mast cells induced by various histamine liberators such as antigen, concanavalin A, ionophore A 23187 and compound 48/80 are significantly diminished in samples incubated with NCO-650.