Clinical and experimental studies with cyclosporine in renal transplantation.
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Azathioprine↗
Biomedical subjects
N Yoshimura
Publications and source records attributed to N Yoshimura.
Impact of the timing of antigen administration on synergistic immunosuppression with cyclosporine.
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Animals↗
[A ventilator system for laser microsurgery of the larynx].
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Adult↗
[A case of cavernous hemangioma of the testis].
Non-germinal cell tumor of the testis is a rare disease, and vascular tumor of the testis is a still rarer disease. Herein, a case of cavernous hemangioma of the testis is reported. A 75-year-old man consulted our department with the complaint of a painless left intrascrotal tumor. Laboratory findings revealed slight leukocytopenia and thrombocytopenia of unknown origin. In chest X-ray, a diffuse reticular shadow was shown and it was considered due to pulmonary fibrosis, but, alpha-fetoprotein and CEA were normal. Left radical orchiectomy was performed under spinal anesthesia. The tumor existed under the tunica albuginea, and the cutting surface of the tumor was brown and irregular. There were hemorrhagic portions in some places. The left epididymis and the left spermatic cord were normal. Histologically, the tumor was diagnosed as cavernous hemangioma. The vascular tumor of the testis is a very rare disease, and only 17 cases have been reported including this case. This case was the 7th case of cavernous hemangioma of the testis, and the first case in Japan. This patient died of respiratory failure due to pulmonary fibrosis and pneumonia. In the autopsy, there was no abnormal finding that was considered to be related to the cavernous hemangioma of the testis.
Aged↗
Transient hyperammonemia during aging in ornithine transcarbamylase-deficient, sparse-fur mice.
Ornithine transcarbamylase [EC 2.1.3.3] (OTC) deficient, sparse-fur (spf) mice are considered to be a good model of human OTC deficiencies. We measured the blood ammonia concentration in spf mice. It was not always at a high level. It was high only during weaning, then gradually decreased and finally fell to a normal level. A similar phenomenon was observed in the change of hepatic ammonia concentration. Excretion of urinary orotate in spf mice was 15-20 times higher than in control mice just after weaning. It also gradually decreased during aging, but remained at a high level compared with adult control mice. OTC activity at pH 7.7 in the liver of spf mice increased significantly between the weaning period and 30-40 days old, when blood ammonia was falling to a normal level.
Aging↗
Comparative specificity and kinetic studies on porcine calpain I and calpain II with naturally occurring peptides and synthetic fluorogenic substrates.
Homogeneous porcine calpain (Ca2+-dependent cysteine proteinase) was found to hydrolyze a variety of peptides and synthetic substrates. Leu-Trp-Met-Arg-Phe-Ala, eledoisin-related peptide, alpha-neoendorphin, angiotensin I, luteinizing hormone-releasing hormone, neurotensin, dynorphin, glucagon, and oxidized insulin B chain were cleaved with a general preference for a Tyr, Met, or Arg residue in the P1 position preceded by a Leu or Val residue in the P2 position. No great difference in specificity was found between low-Ca2+-requiring calpain I and high-Ca2+-requiring calpain II. 4-Methylcoumaryl-7-amide (MCA) derivatives having a Leu(or Val)-Met(or Tyr)-MCA or a Leu-Lys-MCA sequence were also cleaved by either calpain I or calpain II with preference for Leu over Val by a factor of 9 to 16. Calpains I and II showed similar but not identical kinetic behavior for individual substrates. The Km and kcat values ranged from 0.23 to 7.08 mM and 0.062 to 0.805 s-1 for the calpains, while kcat/Km values for the calpains were only 1/433 to 1/5 of those for papain with a given substrate. With succinyl-Leu-Met(or Tyr)-MCA, calpains I and II were half-maximally activated at 12 and 260 microM Ca2+, respectively, and competitively inhibited by leupeptin (Ki = 0.32 microM for I and 0.43 microM for II) or antipain (Ki = 1.41 microM for I and 1.45 microM for II). Thus, this is the first report describing the specificity and kinetics of calpains I and II.
Amino Acid Sequence↗
Calpain and calpastatin in porcine retina. Identification and action on microtubule-associated proteins.
Two forms of Ca2+-dependent cysteine proteinase (calpain, EC 3.4.22.17) and their specific endogenous inhibitor (calpastatin) were partially purified from porcine retina: calpain I (low-Ca2+-requiring form) was half-maximally activated at 8 microM-Ca2+, and calpain II (high-Ca2+-requiring form) at 250 microM-Ca2+. Both calpain I and calpain II were inhibited by calpastatin. Calpain I from porcine retina was shown to be composed of 83 000- and 29 000-Mr subunits, and calpain II of 80 000- and 29 000-Mr subunits, by the use of monospecific antibodies. Calpains I and II were both found to hydrolyse microtubule-associated proteins 1 and 2 rapidly.
Animals↗
Intracellular localization of two distinct Ca2+-proteases (calpain I and calpain II) as demonstrated by using discriminative antibodies.
Intracellular localization of two molecular species of calpain (Ca2+-dependent cysteine proteinase) was studied by immunocyto- and histochemical methods employing antibodies strictly monospecific for the respective antigens. Apparent immunological cross-reactivity between the larger subunits of calpain I (low Ca2+-requiring form) and calpain II (high Ca2+-requiring form) was calculated to be 15-17%, and two steps of affinity chromatography were needed to obtain antibodies which can discriminate between the two proteases. Indirect immunofluorescent staining of cultured PK 15 cells revealed diffuse staining of the cytoplasm with both antibodies against calpain I and calpain II. Preincubation with Ca2+-ionophore had no effect on the staining patterns. Sections of porcine kidney were stained by the avidin-biotinylated peroxidase complex method. The proximal and distal tubules and collecting duct were stained, but the glomerulus, macula densa, and vascular vessels were not stained by either anti-calpain I or anti-calpain II antibodies.
Animals↗
Evidence for membrane-associated calpain I in human erythrocytes. Detection by an immunoelectrophoretic blotting method using monospecific antibody.
Low and high Ca2+-requiring forms of Ca2+-dependent cysteine proteinase are known as calpain I and calpain II, respectively. We have obtained, for the first time, monospecific antibodies for calpain I and for calpain II. Using these antibodies and an electrophoretic blotting method, we have found that a small, but reproducible, amount of calpain I was associated with human erythrocyte membranes while the bulk of the protease was contained in the cytosol. Most of membrane-associated calpain I was extractable with 1% Triton X-100, but not with 0.1% detergent. In the presence of 0.1 mM Ca2+ and 5 mM cysteine, membrane-associated calpain I degraded the membrane protein band 4.1 preferentially and band 3 protein only slowly. The Ca2+-induced autodigestion of the membrane preparation was inhibited by leupeptin but not by a cytosolic calpain inhibitor, calpastatin, added to the incubation medium. No calpain II was detected in either erythrocyte cytosol or membranes when anti-calpain II antibody was used under the same conditions as those for the detection of calpain I.
Antibodies, Monoclonal↗
High arginine levels in intravenous hyperalimentation abrogate post-traumatic immune suppression.
Trauma victims often suffer immune system failure. Oral arginine has strong immune-enhancing properties. The metabolic, hormonal, and immune effects of increasing concentrations of arginine as part of post-trauma intravenous hyperalimentation (IVH) were studied. Groups of 11-14 rats, 275-350 g, underwent jugular vein catheterization and bilateral closed femoral fractures under anesthesia. IVH was started immediately postinjury at a rate of 0.8-1 ml/100 g body wt/hr and continued for 5 days. Twenty percent dextrose and three different amino acid mixtures were given as follows: (A) FreII (1.55 g ARG/1); (B) FreIII (4.05 g ARG/1); (C) modified FreIII (7.9 g ARG/1). All rats lost weight over the 5-day postinjury period; however, rats in groups B and C lost significantly less weight than rats in group A (-3.4 +/- 0.8% of initial body weight and -3.6 +/- 0.9% vs -6.1 +/- 1.2%, P less than 0.05). Rats in group A had negative cumulative nitrogen balance, while those in groups B and C were in highly positive balance. No significant difference in body weight change or nitrogen balance was noted between groups B and C. Trauma-induced thymic involution as assessed by thymic weight and lymphocyte content was greatest in group A, which received the lowest amount of arginine, and was linearly abrogated by increasing the amount of arginine administered (A less than B less than C). Thymocyte immune responsiveness increased with the amount of arginine given as assessed by mitogenesis in response to Con A (stimulation index: A--151.3 +/- 28.8 vs B--243.6 +/- 29.2, P less than 0.01 vs C--321.8 +/- 22.3, P less than 0.001 vs A and P less than 0.02 vs B) and PHA (A--65.0 +/- 14.3 vs B--67.7 +/- 15.3, NS, vs C--117 +/- 14.0, P less than 0.005 vs A and B).(ABSTRACT TRUNCATED AT 250 WORDS)
Amino Acids↗
Immunopharmacodynamic evaluation of cyclosporine-treated renal allograft recipients.
Since the mode of action of cyclosporine (CsA) in man is incompletely understood, there are no monitoring tools to assess immunosuppressive effect in vivo. In vitro CsA inhibits lymphoproliferation in response to allogeneic and mitogenic stimuli, presumably due to reversible suppression of T helper cell generation of interleukin-2. Therefore the present studies examined the immunosuppressive effect of patient sera on a third-party mixed lymphocyte reaction (MLR) as a pharmacodynamic approach to quantify patient responses to CsA administration. Four kinetic patterns of in vitro immunosuppressive activity were discerned: 24/28 (86%) patients showing two cycles of MLR inhibition--namely, a first peak corresponding to absorption of CsA and an independent second peak of immunosuppression (type I), were free of rejection; while 17/23 (74%) patients demonstrating one cycle corresponding to the peak of CsA absorption (type II) suffered rejection episodes (P less than 0.001). In addition, 20 patients generating continuously high levels of in vitro serum activity (type III) were almost all free of rejection, but manifested nephrotoxicity; while two patients showing continuously low levels (type IV) suffered graft loss due to irreversible rejection (P less than 0.01). Thus failure to display either a second peak or continuously high levels of MLR inhibition was associated with a markedly increased incidence of rejection (76% versus 16%). The in vitro functional characteristics of peak-2 were similar to those of CsA, as assessed by the kinetics of inhibition of MLR lymphoproliferation or cell-mediated lympholysis (CML), and by gross chemical properties of partitioning into organic solvents and heat stability. These findings suggest that pharmacodynamic analysis by MLR inhibition not only affords a useful parameter of immunosuppression, but also may provide an in vitro model to dissect the generation and biotransformation of active CsA metabolites.
Cyclosporins↗
Large-scale purification of porcine calpain I and calpain II and comparison of proteolytic fragments of their subunits.
Large-scale purification of calpain [Ca2+-dependent cysteine proteinase; EC 3.4.22.17] from porcine tissues is described. The methods used included chromatographies on DEAE-cellulose, Ultrogel AcA 34, Blue Sepharose CL-6B, and DEAE Bio-Gel A which yielded homogeneous enzyme proteins: 27.0 mg of calpain I (low Ca2+-requiring form) from 5 liters of blood with 17,900-fold purification and 57.6 mg of calpain II (high Ca2+-requiring form) from 1.5 kg of kidneys with 5,800-fold purification. Porcine calpains I and II are half-maximally activated at 2.8 microM and 150 microM Ca2+, respectively. They are composed of large and small subunits: Mr 83,000 and 29,000 for calpain I and Mr 80,000 and 29,000 for calpain II. Gel-electrophoretic analysis of the digest with a-chymotrypsin or Staphylococcus aureus V8 protease revealed that the large subunits of calpains I and II are markedly different in structure whereas the small subunits are most likely identical. Mono-specific antibodies directed toward the respective large and small subunits were used for immunoblotting experiments which established not only the identity among several porcine tissues of calpain I but also that of calpain II. several porcine tissues of calpain I but also that of calpain II.
Animals↗
[Inflammatory vesicosigmoidal fistula. A case report].
Clinical course of a case of inflammatory vesicosigmoidal fistula is presented. The patient, a 44-year-old-male, had chief complaints of pollakisuria and pain upon voiding. Fistula was identified with colon-fiberscopic examination. Primary resection of lesion was performed and postoperative course was uneventful. Histological findings of the specimen were compatible with those of nonspecific inflammation. There was no evidence of neoplastic change. Twenty nine cases of inflammatory vesicosigmoidal fistula were reported in the recent 5 years. It was remarkable that the cases of vesicosigmoidal fistula due to sigmoidal diverticulitis have increased.
Adult↗
[Case of radiation treatment in retroperitoneal hemangiopericytoma].
A 48-year-old woman came to our hospital with complaint of macroscopic hematuria and left lower abdominal pain, on January, 27, 1982. She complained of lower abdominal oppressive pain, but no abdominal tumor was palpated on physical examination. Vaginal examination revealed a stony hard and nodular tumor which was not movable, and as large as a man's fist, on the left side of uterine cervix. IVP revealed left nonfunctioning kidney. Cystoscopy revealed no abnormal finding but left ureteral catheterization could not be done. CTscan revealed intrapelvic homogenous mass which could not be identified from uterus. Pelvic angiography revealed an encasement of the left uterine artery, and moderate hypervascular tumor which deviated the obturatorius artery. Under the diagnosis of retroperitoneal tumor, operation was done on March, 8, 1982. The tumor existed in the retroperitoneal space, and was as large as a man's fist. It was not a movable mass, venous dilatation was found on its surface, and severe adhesion was found between the lateral side of the tumor and the left external iliac artery. So only biopsy was done. Histopathological diagnosis of the specimen was hemangiopericytoma. She received postoperative radiation therapy with total dose 5,000rad in 5 weeks. Now about 2 years have passed, vaginal examination revealed no tumor, and CTscan revealed diminishment of the tumor. Generally radiotherapy is not considered to be effective for hemangiopericytoma, but sometimes it is. Thus preoperative vascular embolization with surgical resection and postoperative radiotherapy or chemotherapy are considered to be necessary for successful treatment of hemangiopericytoma.
Female↗
[A case report of paraurethral leiomyoma in a female].
Paraurethral tumor in females is a relatively rare disease. Herein, a case of paraurethral leiomyoma is reported. A 30-year-old woman consulted our hospital with complaints of a painless tumor and discomfort at the external genital parts. Local examination revealed a thumb's head sized tumor with smooth surface at the anterior wall of the external urethral meatus. Histologically, this tumor contained spindle cells and many collagenous fibers, and was diagnosed as leiomyoma. In Japan, there were 87 cases of female paraurethral benign tumor, which contained 47 cases of leiomyoma including this case. Fifty-four percent of the chief complaints for all female paraurethral tumors were palpable external genital masses; and, palpable painless mass was the most common complaint. Also, the patients with tumors of the posterior or lateral wall complained about urination more frequently.
Adult↗
[A case of leiomyoma of the urinary bladder in a woman with the chief complaint of total urinary retention].
Benign nonepithelial tumor of the urinary bladder is very rare. We experienced a case of a 33-year-old woman who complained of total urinary retention. Vaginal examination revealed a hen's egg sized retrovesical tumor. IVP revealed a filling defect on the cystogram. Cystoscopy revealed protrusion of left side of the bladder neck and intact mucosa. Transvaginal needle biopsy of the tumor was done, and pathohistological diagnosis of the tumor was done, and pathohistological diagnosis of the specimen was leiomyoma. The tumor was intramural type and was resected. It was 5.5 X 5 X 5 cm in size, weighed 80 g, and pathohistological diagnosis was leiomyoma of the urinary bladder. 57 cases of leiomyoma of the urinary bladder including this case have been reported in Japan. No special method of diagnosing leiomyoma of the urinary bladder exists, but in some cases, needle biopsy is very effective.
Adult↗
Identification and quantitative determination of fentanyl metabolites in patients by gas chromatography--mass spectrometry.
Although fentanyl has been used widely as a short-acting narcotic analgesic, its metabolism in humans has not been clarified. In this study, three fentanyl metabolites were identified in the urine of eight surgical patients receiving 0.3-0.5 mg of fentanyl intravenously. The metabolites 4-N-(N-propionylanilino)piperidine, 4-N-(N-hydroxypropionylanilino)piperidine and 1-(2-phenethyl)-4-N-(N-hydroxypropionylanilino)piperidine, and unchanged fentanyl were identified by GC-mass spectrometry in urine collected 6 h after administration. Fentanyl and its main metabolite, 4-N-(N-propionylanilino)piperidine, were determined quantitatively in the urine of five additional patients receiving 0.5 mg fentanyl intravenously. Urinary excretion of fentanyl and 4-N-(N-propionylanilino)-piperidine during the first 12 h after injection accounted for 0.3-4.0% and 26 to 55% of the dose, respectively.
Adult↗
Two distinct Ca2+ proteases (calpain I and calpain II) purified concurrently by the same method from rat kidney.
Two molecular species of calpain (Ca2+-dependent cysteine proteinases) were concurrently purified from rat kidney, both to homogeneity. Calpain I and calpain II having low and high Ca2+ requirements, respectively, were clearly separated on DEAE-cellulose chromatography at pH 7.5, and thereafter they were purified by separate but almost identical procedures which included (NH4)2SO4 fractionation and successive chromatographies on TSK-Gel G 3000 SWG, blue Sepharose CL-6B, and DEAE-Bio-Gel A. The purification folds and activity yields were 6170-fold and 17.8% for calpain I and 4160-fold and 11.9% for calpain II. Ca2+ concentrations for half-maximal activation were 2 microM for calpain I and 200 microM for calpain II. The specific activity of calpain II on casein as the substrate was more than twice higher than that of calpain I. Both enzymes are heterodimers, each composed of 80,000-Da and 25,000-Da subunits. The amino acid compositions of calpain I and calpain II are very similar but not identical. Calpain II is more acidic (pI 4.6) than calpain I (pI 5.3). This paper is the first to describe parallel isolation and characterization of low and high Ca2+-requiring proteases from one single nonmuscular tissue.
Amino Acids↗