Correlation between the pharmacological efficacy of cyclosporine and tacrolimus as evaluated by the lymphocyte immunosuppressant sensitivity test (LIST) and the MTT assay procedure in patients before and after renal transplantation.

OBJECTIVES: Cyclosporine and tacrolimus are calcineurin inhibitors that are used to prevent acute rejection in renal transplant recipients. The lymphocyte immunosuppressant sensitivity test (LIST) can predict the pharmacological efficacy of these immunosuppressive agents for renal transplant recipients. There is a correlation between cyclosporine and tacrolimus pharmacological efficacy as evaluated by LIST by the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay procedure prior to renal transplantation. However, the LIST can also evaluate patients before and after the transplantation. MATERIALS AND METHODS: The present study examined the relationship between cyclosporine and tacrolimus pharmacological efficacy by LIST using the MTT assay in 16 renal transplant recipients at 1, 3 and 12 months after transplantation, as well as before the operation. RESULTS: The relationship of cyclosporine and tacrolimus pharmacological efficacy gave a significant Kendall and Spearman's coefficient correlation in these transplant recipients by the LIST using the MTT assay procedure immediately prior to renal transplantation (rk = 0.711, rs = 0.877, p < 0.01). Furthermore, correlations between the cyclosporine and tacrolimus IC50 values were also observed with a significant Kendall and Spearman's coefficient correlation at 1 and 12 months after transplantation (rk1month = 0.65, rs1month = 0.829, p < 0.01, and k12month = 0.433, rs12month = 0.603, p < 0.01, respectively). However, no statistically significant relationship was observed between the pharmacological efficacies of the calcineurin inhibitors at 3 months after transplantation (rk3month = 0.117, rs3month = 0.1, p > 0.05). CONCLUSIONS: Both cyclosporine and tacrolimus exhibit pharmacological efficacy by the inhibition of calcineurin. However, the correlation between cyclosporine and tacrolimus pharmacological efficacies may be altered, due to immunosuppressive therapy or clinical events at 3 months after renal transplantation.

Cyclosporine pharmacological efficacy estimated by lymphocyte immunosuppressant sensitivity test before and after renal transplantation.

OBJECTIVE: Lymphocyte immunosuppressant sensitivity test (LIST) is useful for predicting the pharmacological efficacy of immunosuppressive agents. In this study, the pharmacological efficacy of cyclosporine was estimated by LIST before and after renal transplantation. METHODS: Lymphocyte immunosuppressant sensitivity test was performed by the 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide (MTT) assay before and at 1, 3, and 12 months after transplantation in 19 consecutive renal transplant recipients. RESULTS: There was wide intersubject variability in cyclosporine IC50 before transplantation [Mean (SD) of 593.9 (1067.6) ng/mL]. This variability worsened 1 month after transplantation [525.7 (1532.7) ng/mL] but decreased at 3 months (193.5 (347.9) ng/mL) and 12 months (75.4 (95.4) ng/mL). In this small study, observed differences in IC50 values for the individual subjects at various time intervals was not associated with the occurrence of rejection, graft loss, and infection episodes. CONCLUSION: Lymphocyte sensitivity to cyclosporine assessed by the LIST assay showed a high level of inter-subject variability particularly before and 1 month after transplantation. The observed difference in IC50 values was not associated with clinical outcome in this small study.

The mouse N-acetylgalactosamine-6-sulfate sulfatase (Galns) gene: cDNA isolation, genomic characterization, chromosomal assignment and analysis of the 5'-flanking region.

Deficiency of lysosomal enzyme N-acetylgalactosamine-6-sulfate sulfatase (GALNS) leads to mucopolysaccharidosis IV A (MPS IV A), for which there is no definitive treatment so far. Although a number of mutations of the GALNS gene of MPS IV A patients have been described, pathogenesis of the disorder still remains elusive. In order to facilitate in vivo studies using model animals for MPS IV A, we isolated and performed molecular characterization of the mouse homolog of human GALNS. The 2.3-kb cDNA contains a 1560-bp open reading frame encoding 520 amino acid residues. The coding region has 84% similarity to the human GALNS cDNA at amino acid level. The mouse Galns gene was mapped by interspecific backcross analysis to the distal region of chromosome 8 where it co-segregates with Aprt. Northern blot analysis showed a wide expression of a single-copy gene, being higher especially in liver and kidney. The Galns gene was isolated from S129vJ genomic library and its genomic organization was characterized. The mouse Galns gene was about 50-kb long and organized into 14 exons and 13 introns. All intron-exon splice junctions conformed to the GT/AG consensus sequence except exon 8/intron 8 junction. Primer extension shows multiple transcription initiation sites between -44 and -75 although major transcription initiation site was observed at -90 bp from the ATG codon. The 5'-flanking region lacks canonical TATA and CAAT box sequences, but is G+C rich with 10 GC boxes (potential Sp1 binding sites), characteristic of a housekeeping gene promoter.

Relationship between the Peripheral Lymphocyte Response to Mycophenolic Acid in vitro and the Level of ATP in Peripheral CD4+ Lymphocytes before and after Renal Transplantation.

OBJECTIVE: The lymphocyte immunosuppressant sensitivity test has been used to predict the pharmacodynamics of immunosuppressive drugs for the purpose of preventing acute rejection and infection after renal transplantation. On the other hand, measuring the ATP levels in peripheral CD4+ lymphocytes is also able to monitor the risks of rejection and infection in transplant recipients. In the present study, we examined the relationship between the mycophenolic acid pharmacodynamics and the ATP levels in peripheral lymphocytes before and after renal transplantation. METHODS: We examined both the pharmacological efficacy of mycophenolic acid and the lymphocyte ATP levels before and 2, 4 and 6 weeks after the operation in 20 renal transplant recipients. The drug's pharmacological efficacy was evaluated by the 50% inhibitory concentration of the drug against the in vitro proliferation of peripheral blood lymphocytes activated by T cell mitogen. The ATP levels in peripheral CD4+ lymphocytes were measured by the Immuknow assay kit. The relationships between the mycophenolic acid pharmacodynamics and ATP levels in peripheral lymphocytes were examined in these recipients. RESULTS: The immunosuppressive effects of mycophenolic acid against mitogen-activated lymphocyte proliferation were significantly and positively correlated with the lymphocyte ATP levels, but only at 6 weeks after transplantation. The relationship was not significant before or at 2 or 4 weeks after the operation. CONCLUSION: Our present data raised the possibility that evaluating the pharmacological efficacy of mycophenolic acid pre-transplantation and monitoring the ATP level 6 weeks after transplantation can predict the risk of rejection and/or infection in renal transplant recipients.

Successful unrelated BMT in a patient with Kostmann syndrome complicated by pre-transplant pulmonary 'bacterial' abscesses.

Kostmann syndrome, severe congenital neutropenia, is often associated with life-threatening bacterial infections. A 5-year-old girl with Kostmann syndrome developed pulmonary abscesses. She was refractory to granulocyte colony-stimulating factor and antibiotics. She underwent unrelated HLA-matched BMT. Myeloablative conditioning consisted of 12-Gy TBI with lung shielding, antithymocyte globulin, etoposide, and cyclophosphamide. After successful engraftment, the pulmonary abscesses resolved by day 75 post-transplant. Although the option of transplantation is not established in the setting of unrelated HLA-matched BMT in Kostmann syndrome, this case may provide useful information. Furthermore, pre-transplant pulmonary bacterial abscesses may not be a contraindication for BMT in some patients with Kostmann syndrome.

Susceptibilities of various myofibrillar proteins to cathepsin B and morphological alteration of isolated myofibrils by this enzyme.

The abilities of cathepsin B purified from liver to degrade purified myofibrillar proteins, myosin, actin, troponin, tropomyosin, and alpha-actinin from rabbit skeletal muscle were studied. The amino acids or peptides liberated from these proteins by cathepsin B were determined quantitatively by fluorometry with o-phthalaldehyde, and qualitatively by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. At a molar ratio of cathepsin B to substrate of 1 : 100, the order of susceptibilities was myosin much greater than troponin greater than tropomyosin much greater than actin. alpha-Actinin was not degraded. Myosin heavy chain was degraded to several fragments with molecular weights of 175,000, 170,000, 160,000, and 145,000, whereas the light chains were scarcely degraded. Cathepsin B degraded troponin-T rapidly, and troponin-I more slowly, but did not degrade troponin-C. Troponin-T and troponin-I were degraded to three fragments with molecular weights of 30,000, 18,000, and 12,800. Tropomyosin was degraded slightly and its product had a molecular weight of 32,000. Actin was also degraded only slowly, and no liberated product could be detected. Morphological changes in myofibrils prepared from glycerinated psoas muscle of rabbit during incubation with cathepsin B were observed. Three notable phenomena were observed: 1) disappearance of the Z-band in the early stage of incubation, 2) disappearance of the M-line following loss of the Z-band, and 3) decrease in the density of the A-band after swelling of the myofibrils.

Effects of cathepsin B, H, and D in pectoral muscle of dystrophic chickens (line 413) of in vivo administration of E-64-c (N-[N-(L-3-transcarboxyoxirane-2-carbonyl)-L-leucyl]-3-methyl-butylamine).

The total activities of cathepsin B and cathepsin H in pectoral muscle of dystrophic chickens (Line 413) were about two times higher than in control chickens (Line 412), and cathepsin D activity was about 3 times higher in this muscle in the dystrophic chickens. When E-64-c, a synthesized potent thiol inhibitor was injected subcutaneously, in various doses, daily for 80 days into dystrophic chickens (L 413), the activities of cathepsin B and cathepsin H were reduced to the levels in control chickens (Line 412), but cathepsin D activity, which is insensitive to E-64-c in vitro, was not changed.

Intra-arterial administration of methotrexate, adriamycin, and cisplatin as neoadjuvant chemotherapy for bladder cancer.

As neoadjuvant chemotherapy for advanced bladder cancer, the intra-arterial administration of methotrexate (MTX), Adriamycin (ADM), and cisplatin (CDDP; IA-MAC) was evaluated. A total of 48 patients with bladder cancer (greater than or equal to T2 or CIS) were selected and received 30.1 mg MTX, 34.5 mg ADM, and 89.1 mg CDDP as an average course. The mean tumor-regression rate after 2 or 3 weeks was 52.3%, and patients with grade 3 transitional-cell carcinoma showed the best results, achieving a 69.6% regression rate. In 30 cases (63%), downstaging was observed. Among the 46 patients who underwent subsequent surgical therapy, the bladder could be preserved in 26 cases by transurethral resection or segmental resection. According to the criteria of the Japanese Association of Cancer Therapy, a histological effect of GIII or better was obtained in 15 cases (29%). The histological effect correlated well with the tumor-regression rate. As compared with intravenous therapy with MTX, vinblastine, ADM, and CDDP (M-VAC), IA-MAC treatment was well tolerated due to its lower degree of bone marrow suppression, and it resulted in a longer disease-free interval and better survival. In addition, the period prior to surgical therapy was shortened in this study. These results suggest that IA-MAC chemotherapy can be useful as an arm of multidisciplinary treatment of advanced bladder tumors.

Mixed connective tissue disease following interstitial cystitis.

A 64-year-old woman complained of severe infrapubic pain and pollakisuria with nausea, vomiting and diarrhea, but with normal urinalysis since 1987. The clinical diagnosis of interstitial cystitis (IC) was made, and cystectomy was performed in 1996. The bladder taken was markedly shrunken with a capacity of 50 ml, and showed bleeding on the mucosal surface. Histological findings of the bladder showed ulcer formation in the mucous membrane, and marked infiltration of mononuclear cells, edema and fibrosis in the submucosal tissue. She had noticed exudative erythema, swelling and sclerosis on the bilateral fingers and dorsal aspects of the hands since 1993, and Raynaud's phenomenon and morning stiffness of the fingers from November, 1998. Laboratory data showed positive anti-nuclear antibody (titer: 1: 2,560) and anti-U1 RNP antibody (titer: 69.5 by ELISA). A diagnosis of mixed connective tissue disease following IC was made.

Organically-bound 3H concentration in rice around atomic energy facilities.

Rice samples collected around atomic energy facilities in the Tokai area were analyzed for organically-bound 3H (OBT). Measurement results were compared with those for samples from control areas in Japan and the People's Republic of China. OBT concentrations of rice in the Tokai area were found to be 4 times higher than the control area as maximum. Although this relatively high 3H concentration shows the effect of 3H release from the facilities into the environment, the concentration level is comparable with those in the rice sample from a northern interior region of China. The OBT concentration of rice sample from the general environment in interior China was 4.5 times higher than the control area in Japan. The radiation dose from the increasing 3H level is negligible in comparison with that from natural radiation.

Reconstructing the history of 14C discharges from Sellafield: part 1--atmospheric discharges.

14C specific activities, above ambient background levels, were determined in individual treerings (corresponding to the years 1950-1999) sectioned from an oak tree that was felled in autumn 1999, from a location 1.5 km east of the Sellafield nuclear fuel reprocessing plant in Cumbria. north-west England. The data were used to produce a new, improved, reconstruction of Sellafield's annual atmospheric 14C discharges between 1951 and 1999, using the most reliable discharge data set (1994-1999) as the primary basis for the determination of a new calibration factor that relates excess 14C activity in individual tree rings to the annual discharge during the corresponding year. The results indicate that the current British Nuclear Fuels plc (BNFL) estimate of total 14C discharges to the atmosphere prior to 1978 is significantly overestimated, while the current estimate of total 14C discharges after 1978 is very similar to that determined in this study. In this study, the total activity of 14C discharged to the atmosphere from Sellafield between 1951 and 1999 is estimated to be 259+/-63 TBq (at 2 std. dev.). The BNFL current estimate is 360TBq.

[Susceptibility testing of clinical strains of Legionella spp. isolated in Japan].

We determined antimicrobial susceptibilities of 23 strains of Legionella spp. clinically isolated in Japan, between September 1994 through November 1999 by using Etest. The MICs of all isolates against macrorides, fluoroquinolones and rifampicin showed 1 microgram/ml or less. Especially rifampicin showed the most potent activity, of which the MICs of all isolates showed 0.064 microgram/ml or less. Although we could not find resistant strains against any antibiotics tested in this study, we think it is important to follow up the susceptibilities of clinical isolates of Legionella.

[A case report of scrotal hemangioma].

A 21-year-old man with painless tumor in the right scrotum was seen at our Department. The tumor was resected surgically and histological examination revealed venous hemangioma. This case is 26th case of scrotal hemangioma in Japan.

[Leiomyoma of the urinary bladder associated with right vesicoureteral reflux].

A 32-year-old female consulted our hospital with the complaint of recurrent urinary tract infections, especially acute pyelonephritis. Cystoscopy revealed a wide-based tumor covered with a normal epithelium at internal meatus and right slight opened orifice, the contraction of which was slightly weak. Excretory urography showed almost normal nephrogram, pyelogram and ureterogram, and voiding cystography revealed right vesicoureteral reflux with grade IIb. Under general anesthesia, tumor resection of the bladder and right ureterovesiconeostomy were carried out. Pathologically the tumor was diagnosed leiomyoma. Right vesicoureteral reflux was speculated to have occurred secondarily to leiomyoma of the urinary bladder.

[Inverted papilloma of the urinary bladder].

Five cases of inverted papilloma in the bladder are reported. All cases were males complaining of dysuria or hematuria. Total cystectomy with construction of ileal conduit was carried out in the first case, and transurethral resection in the other cases. Every surgical specimen was histologically diagnosed as inverted papilloma. The inverted papilloma is a distinct lesion characterized by its rarity compared to frequency of the other vesical papillomas. The prognosis is generally benign, but the possibility of recurrence and malignant transformation indicates the necessity of a close periodical follow-up.

[Ureteral endometriosis].

We present a case of ureteral endometriosis in a 48-year-old postmenopausal patient who was hospitalized complaining of right flank pain & microhematuria. She had no bladder irritability. She had a past history of simple hysterectomy and left oophorectomy because of adenomyomatosis. At that time, right ovary and other pelvic organs were normal. Physical examination revealed slight swelling of the liver without tenderness. Laboratory data at admission revealed severe liver dysfunction due to liver cirrhosis. Excretory urography demonstrated right hydronephroureterosis and severe stricture of the right lower ureter. Right ureteral catheterization demonstrated a high degree of stricture at the distal third. Cytology of right pelvic urine was negative. Right nephroureterectomy was done because long segment of right lower ureter adhered to the peritoneum tightly and we could not rule out ureteral cancer. Pathological diagnosis was extrinsic endometriosis of the right ureter. Ureteral endometriosis is rare and postmenopausal endometriosis is also rare. In our case hyperestrogenemia caused by liver cirrhosis might have played a role in the genesis and progression of endometriosis.

[A case of male duplicated urethra in recurrent urinary tract infection].

A dorsal incomplete duplicated urethra was found in a 13-year-old male, who had noticed occasional urinary incontinence and recurrent urinary tract infection. The accessory urethra lay dorsal from the glans to bladder neck in parallel with the normal ventral urethra. The external orifice of the accessory urethra was also dorsal to the normal urethral orifice in the glans. Voiding cystourethrogram demonstrated double stream but the patient did not notice it because of pseudophimosis. Retrograde urethrogram showed the accessory urethra which arised from the dorsal surface of the prostatic urethra with a parallel normal urethra. The pendular portion of the accessory urethra was surgically removed, the glans portion of the urethra and posterior urethra were cautilized with electrocoagulation for the purpose of preservation of urinary continence. The patient was postoperatively free from urinary tract infection and urinary incontinence.

[Multimodality treatment of adrenal ganglioneuroblastoma: a case report].

We present adult adrenal ganglioneuroblastoma with giant regional lymph node metastasis. The patient was a 21-year-old male, who had a left adrenal tumor diagnosed by ultrasonography incidentally. He was moderately developed and physical examination was almost normal. Blood pressure was within the normal range. Only the value of the vanilmandelic acid, which was 68.7 mg/day, in the urine was abnormal. Other laboratory data were almost normal. Neither chemical nor physical hormonal disorders were present. We carried out the radical surgical treatment by trans-thoracoabdominal approach under the diagnosis of left adrenal neuroblastoma. Pathological diagnosis was ganglioneuroblastoma. After the operation, the patient received both systemic chemotherapy of vincristine and cyclophosphamide and immunotherapy of OK-432 by subcutaneous administration. Furthermore, 40 Gy of the regional irradiation of 60Co to the operative space after the surgery was performed. At eight months after the operation, he is alive with no evidence of disease. Ganglioneuroblastoma is a neuroblastic tumor including neuroblastoma, which commonly occurs in childhood. The prognosis of the neuroblastic tumors which occur either in the adrenal gland of a young child or in an older child is poor. On the other hand, the biological activity of the adult neuroblastic tumors is different from that of the pediatric neuroblastic tumors. In the adult, the adrenal ganglioneuroblastoma does not always have a poor prognosis. The multimodality treatment which we performed is effective for the neuroblastic tumor. Ultrasonography is both effective and adequate for the screening of the retroperitoneal lesion such as in our case.

[Clinical study of serum squamous cell carcinoma-related antigen levels in penile cancer].

A squamous cell carcinoma-related antigen (SCC-Ag), which has been recognized as a tumor marker of cervical cancer of the uterine, was detected in the serum with a radioimmunoassay and its clinical usefulness in penile cancer was evaluated. Seven serum samples of penile cancer, one of Paget's disease and two of condyloma acuminatum were analyzed for the SCC-Ag level. The mean +/- standard deviation of serum SCC-Ag level and positive rate (greater than 2.0 ng/ml) were 1.6 +/- 1.75 ng/ml and 14%. Neither pathological staging nor cell grading of penile cancer showed any relation to the serum SCC-Ag level. When the tumor of the penis has been recognized and a cut-off value of 1.0 ng/ml used, the positive rate of penile cancer was 71.4% and false positive rate of other benign penile tumors was 0%. Serum SCC-Ag values were changed parallel in to the clinical course in three patients. These results suggest that the detection of the serum SCC-Ag value may be of use for detection of penile cancer and monitoring of these patients.

[Ureteritis glandularis: a case report].

A 50-year-old Japanese male hospitalized with the complaint of fever and pyohematuria. An excretory pyelography revealed the right hydronephroureter due to right ureteral stone. When the ureterolithotomy was carried out, a wide-based and rice-sized tumor co-existed at the site of the epithelium of the ureter lithotomized. Resected tumor was pathologically confirmed as poorly differentiated adenocarcinoma with mitosis. Therefore, total nephroureterectomy with bladder cuff resection was done at 10 days after the first operation. However, malignant cells were not found in the surgical specimen or histologically diagnosed localized glandular ureteritis. He is alive without any evidence of recurrence. It was reported that the glandular metaplasia, a relative rare lesion in the ureter, was correlated with carcinogenesis of adenocarcinoma in urothelium. However, when the lesion is small and localized such as in this case it should be treated with ureterectomy and addition of other suitable adjuvant therapies. Furthermore, endourological techniques which have been recently dramatically progressed may become a weapon against this lesion for both treatment and follow-up.