Generation of interspecies limited chimeric nephrons using a conditional nephron progenitor cell replacement system.

Animal fetuses and embryos may have applications in the generation of human organs. Progenitor cells may be an appropriate cell source for regenerative organs because of their safety and availability. However, regenerative organs derived from exogenous lineage progenitors in developing animal fetuses have not yet been obtained. Here, we established a combination system through which donor cells could be precisely injected into the nephrogenic zone and native nephron progenitor cells (NPCs) could be eliminated in a time- and tissue-specific manner. We successfully achieved removal of Six2+ NPCs within the nephrogenic niche and complete replacement of transplanted NPCs with donor cells. These NPCs developed into mature glomeruli and renal tubules, and blood flow was observed following transplantation in vivo. Furthermore, this artificial nephron could be obtained using NPCs from different species. Thus, this technique enables in vivo differentiation from progenitor cells into nephrons, providing insights into nephrogenesis and organ regeneration.

Serial changes in the regional cerebral blood flow of patients with hypertensive intracerebral hemorrhage -- long-term follow-up SPECT study.

We analyzed serial changes in the regional cerebral blood flow (rCBF) of 13 patients with intracerebral hemorrhage by single photon emission computed tomography (SPECT) during the acute- to chronic stage (2 hr to 55 weeks). The (99m)Tc-ethyl cysteinate dimmer ((99m)Tc-ECD) was used as the nuclear mediator. The SPECT timing within 48 hours after the onset was considered to be acute stage, from 48 hours to 4 weeks to be subacute stage, and after 4 weeks to be chronic stage. The region of interest was each hemisphere in the whole brain without ventricles at the thalamic level. For semi-quantitative analysis of rCBF, we used the Brain Uptake Ratio method. Of the 13 patients (mean age 65.5 years), 3 had thalamic-, 4 putaminal-, 5 subcortical-, and one a cerebellar hemorrhage; the hematoma volume varied from 4-50 ml (<20 ml, n=9; 20-30 ml, n=1; >30 ml, n=3; mean 17 ml). The rCBF changes during the long-term follow-up were classified as increase-, decrease-, and unchanged type. Of 5 patients with increased rCBF, 4 made a good recovery and one was severely disabled; of 5 patients with decreased rCBF, 1 made a good recovery, 3 were moderately-, and one was severely disabled. All 3 patients with unchanged rCBF were moderately disabled. Our findings suggest that among patients with hypertensive intracerebral hemorrhage, those with increased rCBF over time may have a favorable outcome. We further need more cases with intracerebral hemorrhage to clarify this trend.

Ischemia-induced neuronal cell death is mediated by the endoplasmic reticulum stress pathway involving CHOP.

Brain ischemia induces apoptosis in neuronal cells, but the mechanism is not well understood. When wild-type mice were subjected to bilateral common carotid arteries occlusion (BCCAO) for 15 min, apoptosis-associated morphological changes and appearance of TUNEL-positive cells were observed in the striatum and in the hippocampus at 48 h after occlusion. RT-PCR analysis revealed that mRNAs for ER stress-associated proapoptotic factor CHOP and an ER chaperone BiP are markedly induced at 12 h after BCCAO. Immunohistochemical analysis showed that CHOP protein is induced in nuclei of damaged neurons at 24 h after occlusion. In contrast, ischemia-associated apoptotic loss of neurons was decreased in CHOP(-/-) mice. Primary hippocampal neurons from CHOP(-/-) mice were more resistant to hypoxia-reoxygenation-induced apoptosis than those from wild-type animals. These results indicate that ischemia-induced neuronal cell death is mediated by the ER stress pathway involving CHOP induction.

Protective effect of sulfobutyl ether beta-cyclodextrin on DY-9760e-induced hemolysis in vitro.

The hemolytic behavior of a novel cytoprotective agent, DY-9760e (3-[2-[4-(3-chloro-2-methylphenyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1-(4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate) was investigated using rabbit erythrocytes. Further, the effects of water-soluble cyclodextrin derivatives, such as 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) and sulfobutyl ether of beta-cyclodextrin (SBE-beta-CyD), on the hemolytic activity of DY-9760e were studied. DY-9760e induced hemolysis at concentrations >0.2-0.3 mM in phosphate buffered saline (PBS) of pH 4.0 and 6.0, where DY-9760e is predominantly in dicationic and monocationic forms, respectively. The hemolytic activity of the monocationic DY-9760e was higher than that of the dicationic species, and the hemolysis at pH 4.0 involved the formation of methemoglobin. DY9760e induced the morphological change of erythrocytes towards membrane invagination at both pH 4.0 and 6.0. SBE7-beta-CyD significantly suppressed the DY-9760e-induced hemolysis and morphological change at both pH 4.0 and 6.0, as well as the formation of methemoglobin at pH 4.0. On the other hand, HP-beta-CyD suppressed only the hemolysis, but neither the morphological change nor the formation of methemoglobin. In addition, the inhibitory effect of SBE7-beta-CyD on the hemolysis was greater than that of HP-beta-CyD. The superior inhibitory effect of SBE7-beta-CyD on the DY-9760-induced hemolysis, the morphological change, and the formation of methemoglobin may be attributable to the formation of a stable inclusion complex with DY-9760e and to the weaker hemolytic activity of SBE7beta-CyD than HP-beta-CyD. These results suggest potential use of SBE7-beta-CyD as a parenteral carrier for DY-9760e.

[Dynamic analysis of airway temperature in dogs breathing cold air].

Exercise in cold, dry air induces bronchoconstriction. However, the time course of airway cooling during the breathing of cold air has not been investigated. In this study in dogs, the temperatures of tracheal gas and the tracheal wall were measured continuously while the animal was breathing cold air at approximately 4.5 degrees C. The temperature of the tracheal gas decreased during the inspiratory phase, increase slightly early in the expiratory phase, and then decreased to the level at end-expiration. The lowest temperature of the tracheal gas decreased significantly, from 29.7 +/- 2.4 degrees C (mean +/- SD) to 25.7 +/- 2.8 degrees C. The highest temperature also changed significantly, but the decrease was less than 1 degree C. The lowest temperature of the tracheal wall decreased from 31.1 +/- 2.6 degrees C to 30.5 +/- 2.5 degrees C during cold-air breathing, but the difference was not significant. When a dog inspired deeply during a cough, the temperature of tracheal gas did not decrease linearly with the progression of inspiration. However, the decrease in the tracheal wall temperature was almost directly proportional to inspiratory volume. We concluded that both the tracheal gas and the tracheal wall temperatures were resistant to cooling with cold-air breathing. It remains uncertain whether rapid, deep breathing during exercise decreased the tracheal wall temperature.

Improvement of subcutaneous bioavailability of insulin by sulphobutyl ether beta-cyclodextrin in rats.

The objective of this study was to examine and compare how hydrophilic beta-cyclodextrin derivatives (beta-CyDs) improve the bioavailability of insulin following subcutaneous injection of insulin solution in rats. When insulin solutions in the absence of beta-CyDs were injected into the dorsal subcutaneous tissues of rats, the absolute bioavailability of insulin calculated from plasma immunoreactive insulin (IRI) levels was approximately 50%. When maltosyl-beta-cyclodextrin was added to the solutions, there was no change in the plasma IRI levels and hypoglycaemia compared with those of the insulin-alone solution. Dimethyl-beta-cyclodextrin decreased the bioavailability of insulin, although it increased the maximal concentration of IRI in plasma and the capillary permeability of the fluorescein isothiocyanatedextran 40, a non-degraded permeation marker. When insulin solutions containing sulphobutyl ether-beta-cyclodextrin with a degree of substitution of the sulphobutyl group of 3.9 (SBE4-beta-CyD) were injected, the IRI level rapidly increased and maintained higher IRI levels for at least 8 h. The bioavailability of the insulin/SBE4-beta-CyD system was about twice that of insulin alone and approached 96%. The enhancing effects of SBE4-beta-CyD may be in part due to the inhibitory effects of SBE4-beta-CyDs on the enzymatic degradation and/or the adsorption of insulin onto the subcutaneous tissue at the injection site, although this does not apparently facilitate capillary permeability. These results suggest that SBE4-beta-CyD in aqueous insulin injection for subcutaneous administration is useful for improving the bioavailability and the hence the pharmacological effects of insulin.

[A study of ambulatory treatment for pulmonary tuberculosis in foreigners residing in Japan].

We studied 130 cases of pulmonary tuberculosis in foreigners residing in Japan to obtain the results as follows; 1. Of the cases of pulmonary tuberculosis in foreigners who are registered and receiving treatment in Japan, 20.3% were treated at three dispensaries of the Japan Antituberculosis Association in Tokyo. 2. The nationality of the cases treated was China in more than half of them, followed by the Republic of Korea. 3. The number of days taken from entry into Japan to the start of treatment was about 11.4 months; 0.9% of the total number of cases examined by chest radiophotography required medical treatment. 4. Their living conditions in Japan according to questionnairing are: 56.2% have jobs in Japan; working hour, 4.99 +/- 1.19 hours a day; 64.4% take night work; 57.6% work in food/drink service industry; living space is 12.5 m2; 52.4% share the same house with other persons, living together with 1.6 persons. 5. As for the type of illness at the start of treatment, GAKKAI classification type III accounted for 90% and spread 1 83.8%. GAKKAI classification type II accounted for 10%, consisting of many relatively mild cases. 6. The defaulter rate was high at 40.8%. The reason for defaulting was broken down to discontinuation on his own 68%, repatriation 15% and side-effects 19%. The time to default was average 3.2 +/- 3.1 months after the start of treatment. They defaulted 1.2 +/- 0.4 times on the average. 7. To reduce the defaulter rate to the minimum in treating the foreigners residing in Japan, the following may be needed. a. To give guidance on the regimen including the need of treatment and risk associated with discontinuation of treatment at the first visit. b. Measures to reduce the amount to be born by the individual in the medical expenses. c. Preparation of a pamphlet for therapeutic guidance in foreign languages.

Methotrexate, vinblastine, doxorubicin, and cisplatin for advanced urothelial cancer.

A series of 31 patients with advanced urothelial cancer were treated with combination chemotherapy consisting of 1-4 cycles of methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC). Of the 31 patients, 29 had measurable and evaluable lesions. A complete remission was achieved by 4 of these 29 patients (14%) for 1-46 months. A partial remission was observed in 14 of the 29 patients (48%) for 1-9 months. Whereas bony and hepatic metastatic lesions did not respond, some nodal (7/12), pulmonary (4/8), and pelvic lesions (2/3) as well as primary bladder tumors (4/6) and a tumor marker (1/2) responded. Complete tumor remission was observed in nodal (2/12) and pulmonary (1/8) metastatic lesions, in invasive lesions to the prostate and seminal vesicle (1/1), and in primary lesions in the bladder (2/6), ureter (1/1), and urethra (1/1). Two of three patients with non-transitional cell tumors attained a partial remission for 1-7 months. Complete remission of the pulmonary lesions was obtained in a case of squamous cell cancer of the bladder with pulmonary metastases. The toxicity of this regimen was generally tolerable and included moderate to severe myelosuppression, mild to moderate nausea and vomiting, renal toxicity, and mucositis. These results suggest that the M-VAC regimen holds promise for the treatment of advanced metastatic transitional cell cancer as well as non-transitional cell cancer of the urothelium.

[Clinical observation of twelve cases of testicular cancer].

Twelve cases of testicular cancer were treated at our Department of Urology between 1984 and 1985. The mean age of the patients was 35.2 years old. Eight of them were pure seminoma in histology, other cases were non-seminomatous germ cell tumors (NSGT). Seven cases were of stage I testicular cancer, 3 cases were of stage IIA, and 2 cases were of stage IIB. Patients with stages I and IIA seminoma were given radiation to the ipsilateral iliac and bilateral paraaortic caval nodes to the crura of the diaphragma. Two patients with stage IIB seminoma were treated with PVB therapy and retroperitoneal lymph node resection. Chemotherapy containing CDDP was effective to reduce the tumor size. Two cases were of stage I NSGT, one case was treated with high orchiectomy only, and the other case was treated with chemotherapy after high orchiectomy. Chemotherapy and retroperitoneal lymph node resection were given to two patients with stage IIA NSGT. Abnormally elevated values of lactate dehydrogenase were observed in cases with a primary tumors weight larger than about 100 g. Six of the 8 cases of pure seminoma had slightly elevated beta-human chorionic gonadotropin levels. Two cases with bilateral testicular cancer were found among the 12 cases so that necessity of meticulous palpation of remaining testis was felt. Recurrent change was not observed in any of the 12 cases so that testicular cancer is considered to be an almost curable disease.

[Rhabdomyosarcoma of the prostate in a child: report of a case].

A case of prostatic rhabdomyosarcoma in a 5-year-old boy is presented. He was referred to us because of complete urinary retention. Histologically, embryonal rhabdomyosarcoma of the prostate was found. Initially, he was treated with combination chemotherapy consisting of vincristine, actinomycin-D, adriamycin and radiation therapy. This therapy reduced the size of the tumor markedly. Therefore, we performed prostatectomy. No tumor cells were found on the surgical margin of the resected prostate. About 11 months after the operation, rapidly growing recurrent lesions were found near the bladder neck. Bilateral hydronephrosis and an intravesical massive space occupying lesion were found on the excretory pyelograms and cystogram. Then three drug combination chemotherapy consisting of vinblastine, cis-diamminedichloroplatinum and bleomycin (PVB therapy) was administered and the first course of the chemotherapy reduced the size of the recurrent tumor. Drainage of contrast medium from bilateral kidneys became smooth, and no gross hematuria or severe frequency was observed. Total cystectomy with ileal conduit urinary diversion was performed. Four months after this operation, metastatic lesions appeared at the pubic bone, right ischiadic bone and rectum, and they were resected operatively. However, multiple pulmonary metastases soon occurred, and he did on June 30, 1985, approximately 2 years after the first diagnosis. We have discussed the effectiveness of combination chemotherapy, especially PVB therapy for recurrent cases.

[Combination chemotherapy with adriamycin, carboquone and 5-Fu for bladder cancer].

We treated 25 bladder cancer patients with combined cytotoxic chemotherapy of adriamycin (10--20 mg/day administration on days 1, 2 and 3), carboquone (4 mg/day administration on day 1 of weeks 3, 4 and 5 and 5-Fu (200 mg/day for 5 weeks) or futraful suppositories (750 mg/day for 5 weeks) as one course. According to the Koyama - Saitoh criteria, CR + PR was observed in 4 (17.4%) of 23 patients excluding the 2 dropout patients. According to Karnofsky's criteria, an effect with chemotherapy was observed in 9 (39.1%) of the 23 patients. There was a relatively good response rate in a group of 12 patients with the superficial tumors as compared with a group of 11 patients with the deep tumors. There were no severe adverse reactions.

Polymorphonuclear leukocyte chemotaxis in sporotrichosis.

A lesion of sporotrichosis shows a characteristic arrangement of the infiltrate in three zones: the central suppurative zone composed of polymorphonuclear leukocytes, the tuberculoid zone and the round cell zone. For investigating the pathomechanism of the suppurative zone formation, polymorphonuclear leukocyte chemotaxis to sporotrichin was assayed with the agarose plate method. Polymorphonuclear leukocytes in the patients with sporotrichosis showed an enhanced chemotactic index in comparison with that in the control subjects. However, there was no difference in non-specific chemotaxis to bacteria-derived substance between the patients and the control subjects.