Long-term hepatic allograft acceptance based on CD40 blockade by ASKP1240 in nonhuman primates.

Blockade of the CD40-CD154 costimulatory signal is an attractive strategy for immunosuppression and tolerance induction in organ transplantation. Treatment with anti-CD154 monoclonal antibodies (mAbs) results in potent immunosuppression in nonhuman primates (NHPs). Despite plans for future clinical use, further development of these treatments was halted by complications. As an alternative approach, we have been focusing on the inhibition of the counter receptor, CD40 and have shown that a novel human anti-CD40 mAb, ASKP1240, markedly prolongs renal allograft survival in NHPs, although allografts eventually underwent chronic allograft nephropathy. On the basis of our previous findings that a CD40-CD154 costimulation blockade induces tolerance to hepatic, but not cardiac, allografts in rodents, we tested here our hypothesis that a blockade of CD40 by ASKP1240 allows acceptance of hepatic allografts in NHPs. A 2-week ASKP1240 induction treatment prolonged liver allograft survival in NHPs; however, the graft function deteriorated due to chronic rejection. In contrast, a 6-month ASKP1240 maintenance monotherapy efficiently suppressed both cellular and humoral alloimmune responses and prevented rejection on the hepatic allograft. No serious side effects, including thromboembolic complications, were noted in the ASKP1240-treated monkeys. We conclude that CD40 blockade by ASKP1240 would be a desirable immunosuppressant for clinical liver transplantation.

A notable case report of May-Hegglin anomaly with immune complex-related nephropathy: a genetic and histological analysis.

May-Hegglin anomaly (MHA) is a rare autosomal dominant disease characterized by macrothrombocytopenia and leukocyte inclusions with microfilaments in the ribosomes. Mutations in the MYH9 gene, encoding non-muscle myosin heavy chain IIA (NMMHC-IIA) have been identified in patients with MHA and other MYH9-related diseases. Two young males (an older and younger brother) presented with macrothrombocytopenia and leukocyte inclusion bodies. Electron microscopy (EM) revealed parallel filaments in leukocyte inclusion bodies characteristic of MHA. Immunofluorescence microscopy (IF) showed NMMHC-IIA antibodies in 1 - 2 leukocyte inclusion bodies. These findings were consistent with MHA and they were identified to express the MYH9 mutation, D1424H. The older brother underwent a renal biopsy because of persistent proteinuria. Histology revealed mesangial proliferative glomerulonephritis with granular deposits of IgG and C1q. EM showed that the dense deposits were located in subendothelial cells, mesangial cells and Bowman's capsule. Immunocytochemistry revealed that NMMHC-IIA antibodies were localized in podocyte and endothelial cells in the glomerulus. Moreover, the expression of nephrin and podocin, slit diagram protein, was normal. An inflammatory mechanism may occur separately from MYH9-related disease. This report presents a case of MHA with immune complex-related nephropathy.

14-3-3ζ-Mediated Stimulation of Oxidative Phosphorylation Exacerbates Oxidative Damage Under Hypothermic Oxygenated Conditions in Human Renal Tubular Cells (HK-2).

Cellular survival and death are at least partially regulated by the phosphorylation of proteins. A chaperon protein, 14-3-3ζ, regulates the activity of many proteins by covering the phosphorylation site within a 14-3-3 binding motif. Therefore, regulation of 14-3-3ζ activity may affect the fate of cells subjected to cold preservation and/or hypothermic oxygenated conditions. The present study assessed whether 14-3-3ζ protects cells from hypothermic oxygenation-induced injury and clarified its role in mitochondrial functions. Human renal tubular cell line HK-2 or 14-3-3ζ-overexpressed HK-2 (ζHK-2) cells were subjected to 72 hours of normoxic cold preservation in UW solution with or without antioxidants and hydroperoxides. Cellular death, adenosine triphosphate (ATP) content, and MTT catabolism were evaluated. Deferoxamine treatment reduced cellular death and augmented ATP content in both cell types. These indices were higher in ζHK-2, regardless of deferoxamine treatment. Exposure to hydroperoxides did not affect cellular death in either cell type, whereas hydroperoxide supplementation significantly reduced ATP content, except for low-dose hydrogen peroxide in HK-2 cells. MTT assay at normal state showed higher values in ζHK-2 cells, whereas it was impaired by hydroperoxides in both cell types. These results suggest that accumulation of hydroperoxides as a byproduct of the augmented oxidative phosphorylation by 14-3-3ζ overexpression causes mitochondrial dysfunction. In conclusion, despite possessing many potentially protective functions, 14-3-3ζ exacerbates cellular injury under hypothermic oxygenated conditions. 14-3-3ζ accelerates mitochondrial functions together with iron-dependent oxidative damage. Although further investigations are necessary, upregulation of 14-3-3ζ could be a method to maintain mitochondrial function under hypothermic oxygenated conditions, as shown in hypothermic machine preservation of renal grafts, when appropriate antioxidant treatment is administered.

Development of ultradian rhythm of EEG activities in premature babies.

Ultradian rhythms in electroencephalogram (EEG) activity in premature neonates were studied. The values of square roots of total power (SQP) of EEG showed a normal Gaussian distribution. We analyzed the smoothed time series of SQP by autoregressive analysis and component analysis. Ultradian rhythm was divided into four groups by period length (group I: 5-30 min, group II: 30-60 min, group III: 60-100 min, group IV: longer than 100 min). Group IV rhythms were not detected before 32 weeks of gestation. Changes in the characteristic values of ultradian components in group II showed that there were critical periods in the development of central nervous activity at 29 and 33 weeks of gestation. This method allowed detection of ultradian rhythms, even at very early gestation, directly from the neonatal central nervous system. This method may be applicable to ill infants.

[Urinary cytology studied during 10 days after transurethral resection of bladder tumour and its relation to tumour recurrence].

Whether or not recurrence is related to the results of urinary cytology examined within 10 days after transurethral resection of bladder tumour was studied retrospectively in 47 patients with superficial bladder tumour. Of 7 cases with positive cytology during the postoperative 10 day period, 4 cases had a later recurrence of tumour and 2 cases had a residual tumour due to incomplete resection of original tumour. By microscopic chromocystoscopy, in 11 patients concurrent urothelial atypia (carcinoma in situ or dysplasia) was found in the apparently normal mucosa. Nine of the 11 cases had a later recurrence of tumour or a residual tumour. Of in total 15 patients combined with abnormal cytology and concurrent urothelial atypia, 12 (80%) were found with recurrence of tumor cystoscopically 4 approximately 64 months (mean; 20.6 months) after TUR. This recurrence rate was significantly (p less than 0.05) higher than that (42.4%) in patients without positive cytology and concurrent urothelial atypia. These results suggest that positive urinary cytology during 10 days subsequent to TUR and/or association with concurrent urothelial atypia are indicative of later tumour recurrence, incomplete resection of tumour or presence of additional occult tumour foci.

[Hairy cell leukemia expressing SIgM+, SIgG-, CD11b+ and CD21+ and accompanying lymphadenopathy without splenomegaly].

Hairy cell leukemia (HCL) expressing both surface monocytoid antigen and IgM (kappa) was reported. A 62-year-old male was admitted to our hospital in September 21, 1989 because of leukocytosis. Physical examinations showed axillary and inguinal lymphadenopathy but no hepato-splenomegaly. The leukocyte count was 12,600/microliters with 73% of abnormal cells like large lymphocytes which had abundant cytoplasm and hairy appearance under phase microscopy. They had ruffles with microvilli under electron microscope. Bone marrow puncture showed normocellular marrow with 71.2% of abnormal cells similar to the peripheral blood. Surface markers were CD11b+, CD21+, HLA-DR+, Tac- and IgM (kappa). They were positive for ++acid phosphatase staining, but negative for peroxidase and tartrate-resistant acid phosphatase staining. He was diagnosed as Japanese type HCL. HCL expressing both surface monocytoid antigen and IgM is rare and the clinical features of our case are compared with those reported in Japan.

[Quantification of electroencephalogram continuity by 24-hour recording in very premature infants].

We aimed to quantify electroencephalogram (EEG) continuity for 24 hours in very premature infants. A total of 122 days of continuous two-channel EEG recordings were performed in 28 premature infants from 26 to 33 weeks of conceptional age (CA). None of the infants showed any evidence of neurological impairments during the course of their hospitalizations and showed normal neurological outcome. The 24-hour EEG recording was divided into 5.5-minute periods. The EEG of each period was classified into five EEG categories according to EEG continuity, and the percentage of each of them during the 24-hour recording was calculated. The percentages of continuous and continuous dominant EEG categories were increased with increasing CA. On the other hand, the percentages of discontinuous and discontinuous dominant EEG categories were decreased with increasing CA.

[An application of autoregressive model to pattern discrimination of brain electrical activity mapping].

We developed a method of illustrating significant regional differences between two records of brain electrical activity mapping (BEAM), using pattern discrimination of autoregressive (AR) EEG analysis. In this study, the EEG data could be objectively reduced to a few AR coefficients, and this statistical method was successfully applied to discrimination of differences in BEAM among records from two or more subjects. A significant difference was illustrated at the right parietal region between two age groups, 6 years and 8 years, in normal children. It was concluded that this new approach of BEAM analysis through a statistical taxonomy was clinically useful for evaluation of the normal development of BEAM in children and for localization of functional brain abnormalities.

[Relative size of the ventricle to the hemisphere in the neonatal MRI].

Using a software program for measuring surface area, we quantified the relative size of four parts of the lateral ventricles, including the body, the trigone, the anterior horn and the occipital horn, compared to the hemispheres in the axial plane of magnetic resonance imaging (MRI) in neonates. In 44 neonates without any neurological disorders from 26 to 41 weeks of gestational age (GA), MRI was performed between 12 and 124 postnatal days. The mean of the relative size of the ventricle compared to the hemisphere (RSVH) among the subjects showed a significant left-right asymmetry that was observed only in the occipital horn. However, in the body, the anterior horn and the occipital horn, the percentage of neonates with a larger left RSVH compared to the right RSVH was significantly higher than the percentage of neonates with a larger right RSVH compared to the left RSVH. The RSVH of the body and the occipital horn increased according to the number of postnatal days and decreased according to GA. Measuring RSVH was useful in assessing the size of the ventricle in the axial plane of neonatal MRI. It clarified the fact that normative asymmetry, GA of subject, and the number of postnatal days should be considered in assessing the size of ventricle.

[Developmental characteristics of the alpha waves of EEG in normal healthy children].

The records of waking state electroencephalography (EEG) of 1,340 normal children aged from 3 to 12 years were subjected to analysis using an autoregressive model. The results were summarized as follows. (1) 6,814 components of second-order activities were obtained through component analysis of the 1,218 EEG records. The frequency polygons of damping frequency of the second-order component waves verified several modes, each of which was enhanced in the eight frequency ranges. (2) The average percent-power of the alpha-1 waves decreased with increasing ages from 3 years, whereas that of the alpha-2 waves increased. That of the alpha-3 waves showed no change with ages. (3) The average damping times of alpha-2 waves increased with increasing ages. (4) Dominancy in frequency showed a course of two step increments at 3-6 and 10 years of age. The former was chiefly due to the increase in the alpha-1 wave and the latter in the alpha-2 wave. There were no significant increases of alpha-3 waves. (5) By means of EEG pattern discrimination, the differences in alpha components among different age groups were obtained as generalized (Maharanobis') distances. The alpha-1 activity showed a significant difference in amount between the age group of 3-6 years and the other age groups. The alpha-2 activity showed a significant difference in amount between the age group of 10 years and the other age groups. (6) The development of brain activity proceeds faster in the posterior areas and relatively slower in the central areas.

[Developmental characteristics of the beta waves of EEG in normal healthy children].

By the use of an autoregressive (AR) model, development of beta-waves in quantitative EEG parameters was studied in 1,340 normal children aged from 3 to 12 years. The results were summarized as follows. The beta band(13.5-25 Hz) was divided into three frequency ranges; beta-1(13.5-17.4 Hz), beta-2(17.5-21.4 Hz) and beta-3 (21.5-25 Hz) waves. The average power and percent-power of the beta-1 and beta-2 waves increased with advancing age, whereas these of the beta-3 wave showed no change with advancing age. By means of EEG pattern discrimination, critical period of beta band in the developing EEG was observed at the age of 4 years in frontal region, 10 years in central region and 3 to 7 years in occipital region. Those results suggest that the development of beta activity may be faster in frontal, occipital areas and relatively slower in central area.

[Enhancement of the muscle relaxant action of baclofen by glucocorticoids].

Effects of pretreatment with hydrocortisone or prednisolone on the pharmacological effects of baclofen, particularly on the muscle relaxant action, were studied in young adult rats. Muscle relaxation was determined using an inclined board (50 degrees). A single injection of hydrocortisone (10 mg/kg, s.c.) 15 min prior to the administration of baclofen (5 mg/kg, i.p.) increased significantly the muscle relaxant time by baclofen. Prednisolone pretreatment (2 mg/kg, s.c.) also prolonged baclofen-induced muscle relaxation. Muscle relaxation due to tolperisone (50 mg/kg, i.p.) and mephenesin (80 mg/kg, i.p.) administration was enhanced by the pretreatment of hydrocortisone. Hydrocortisone pretreatment enhanced the muscle relaxant time of diazepam (2 mg/kg, i.p.) but at a lesser extent and only in males. No synergistic effect of hydrocortisone on the muscle relaxation time by suxamethonium (0.01 mg/kg, i.p.) was observed. Seven daily injections of 2 mg/kg hydrocortisone as well as of 0.4 mg/kg prednisolone did not enhance the muscle relaxation by baclofen when tested 24 hr after the last injection of the two steroids. Fifteen min priming with hydrocortisone had no effect on the antinociceptive and hypothermic effects of baclofen. 3H-Baclofen was injected i.v. as a tracer of 1 mg/kg unlabeled baclofen. In hydrocortisone-treated rats, the uptake of 3H-baclofen into the hippocampus, medulla oblongata and cerebellum was significantly higher than that in the vehicle-treated rats, whereas the radioactivity in the cervical, thoracic and lumbar spinal cord tended to decrease. The results indicate that acute priming with hydrocortisone enhances rather selectively the muscle relaxant action of the centrally acting muscle relaxant drugs. It is suggested that the primary site of the synergistic action of hydrocortisone to baclofen could be at the supraspinal level of the CNS.

Increased transport of theophylline into gastrointestinal lumen and gastrointestinal dialysis by activated charcoal in rats with hepatic cirrhosis.

The characteristics of exsorption and/or excretion of theophylline into the small intestinal lumen in rats with hepatic cirrhosis (HC rats) induced by carbon tetrachloride were investigated by an in situ single-pass perfusion technique. The serum concentrations of theophylline after i.v. administration of aminophylline (10 mg/kg) in the HC rats were significantly higher than those in normal rats during the experimental period. Moreover, the exsorption of theophylline from blood into the intestinal lumen was significantly increased in the HC rats compared with the normal rats. Treatments with oral activated charcoal reduced the serum theophylline levels in the HC rats. Consequently, gastrointestinal dialysis by oral administration of activated charcoal may be a useful method to remove poisonous drugs from the blood in patients with hepatic failure (including cirrhosis), which decreases the systemic clearance.

Sequential changes in electroencephalogram continuity in very premature infants.

This study was conducted to quantify sequential changes in electroencephalogram (EEG) continuity for 24 h in very premature infants. For a total of 122 days, continuous 2-channel EEG recording was conducted for 28 premature infants from 26 to 33 weeks of conceptional age (CA). None of the infants showed evidence of neurological impairment during hospitalization. Normal neurological outcome was noted at a minimum 12 months of age. By classifying each 5.5 min epoch according to EEG continuity, the number of contiguous epochs of each series of discontinuous type (DTs) and the number of epochs between two series of discontinuous type (IDTIs) were counted at each CA. The duration of DT decreased with increasing CA. The mean duration remained at 13-16 min after 29 weeks CA. The mean duration of each IDTI increased with CA, up to about 1 h at 33 weeks. A constant period of DTs was noted at longer intervals with increasing CA. These changes appear related to the development of sleep state organization with CA.