Preservation of pancreas in the University of Wisconsin solution supplemented with AP39 reduces reactive oxygen species production and improves islet graft function

Ischemia-reperfusion injury (IRI) results in increased rates of delayed graft function and early graft loss. It has recently been reported that hydrogen sulfide (H₂S) protects organ grafts against prolonged IRI. Here, we investigated whether the preservation of pancreas in University of Wisconsin (UW) solution supplemented with AP39, which is a mitochondrial-targeted H₂S donor, protected pancreatic islets against IRI and improved islet function. Porcine pancreata were preserved in the UW solution with AP39 (UW + AP39) or the vehicle (UW) for 18 h, followed by islet isolation. The islet yields before and after purification were significantly higher in the UW + AP39 group than in the UW group. The islets isolated from the pancreas preserved in UW + AP39 exhibited significantly decreased levels of reactive oxygen species (ROS) production and a significantly increased mitochondrial membrane potential as compared to the islets isolated from the pancreas preserved in the vehicle. We found that the pancreas preserved in UW + AP39 improved the outcome of islet transplantation in streptozotocin-induced diabetic mice. These results suggest that the preservation of pancreas in UW + AP39 protects the islet grafts against IRI and could thus serve as a novel clinical strategy for improving islet transplantation outcomes.     

Monoclonal antibodies against human erythrocyte membrane antigens and their reactivities with hematopoietic cells

Four monoclonal antibodies against human erythrocyte membrane antigens were established. The antigenic determinants of KOR-E1, E3, E6 were Pr1h antigen, Wrb antigen, and the trypsin sensitive portion of glycophorin A (EnaTS) respectively. The antigen recognized by KOR-E4 could not be determined. The reactivities of these antibodies with normal hematopoietic cells, malignant hematopoietic cell lines (N = 31), and fresh leukemic cells obtained from 128 patients with various types of leukemias were studied. All antibodies reacted only with erythrocytes among peripheral blood cells, and also KOR-E6 reacted only with erythroid cells among bone marrow cells. KOR-E3 had no reactivity with any cell lines examined, and KOR-E1 and KOR-E4 were reactive with some lymphoid cell lines. However, KOR-E6 had specific reactivities with erythroid (HEL, K562), megakaryocytic (CMK-1), multiphenotypic (KOPM-28), and basophilic (KU-812) cell lines. The antigen (glycophorin A) recognized by KOR-E6 was expressed on a small population of mononuclear cells separated from acute lymphoblastic leukemia (3/70), acute myelogenous leukemia (2/12), monosomy 7-myeloproliferative disorder (1/1), juvenile CML (1/1), and transient myeloproliferative disorder with Down's syndrome (4/12), although it could not be determined whether these cells were leukemic cells or not. KOR-E6 was reactive with a large population of leukemic blasts in erythroleukemia (2/2) and acute megakaryoblastic leukemia (3/6). Thus, KOR-E6 appears to be an erythroid marker of leukemic cells.     

Clinical significances of intraoperative radiation therapy for aged patients with bladder cancer]

Seven elderly patients, 79-88 years old, with bladder cancer were treated by transvesical tumorectomy with intraoperative radiation therapy (IORT). The cancers appeared to be of high grade and high stage by cystoscopy and other examinations, and consequently they were diagnosed to be over stage T2. Therefore, all patients were thought to be candidates for total cystectomy. But their ages and complications precluded this treatment, so we decided to carry out the "palliative" IORT. The operation of IORT required less than two hours and required less than 200 ml of blood loss. There were no complications such as hematuria, irritable bladder, and rectal symptoms. The postoperative stage diagnoses coincided with the preoperative ones in 5 cases, but two cases were overdiagnosed. Five patients died after more than one year and 11 months, but four patients died due to other diseases, without cancer. One patient died due to pulmonary cancer confirmed by autopsy. Recurrence was seen in one case. These results confirmed that IORT was effective for local control of bladder cancer and partially prophylactic for recurrence. Furthermore, this treatment seemed to be even curative for some cases. We recommend this modality of treatment for some of aged patients and patients with complications who are unable to undergo cystectomy.     

A comparative study of measurement of arterial blood pressure using HEM-802F and arterial cannulation

Finger arterial blood pressures determined by a newly developed sphygmomanometer, HEM-802F, were compared with arterial pressure obtained from direct measurement of the radial artery. An excellent correlation was found between the two methods (systolic: r = 0.93, diastolic: r = 0.91), although there was a large variability among individual subjects. The range of differences between them are from +32 to –13mmHg for systolic and +15 to –25mmHg for diastolic blood pressure measurement. HEM-802F underestimated systolic pressure (–4.0mmHg) and overestimated diastolic pressure (+6.7mmHg), compared with intra-arterial readings.The HEM-802F was useful for the non-invasive arterial pressure monitoring during general anesthesia.(Shigemi K, Takahashi H, Hashimoto S et al.: A comparative study of measurement of arterial blood pressure using HEM-802F and arterial cannulation. J Anesth 4: 91–93, 1990)     

P53 and rb alterations in operable breast-cancer

We analyzed association between p53 and/or Rb expression and clinicopathologic variables or Ag-NOR counts, and then ascertained whether p53 and/or Rb expression would be useful for estimating prognosis in 81 breast cancer patients. Positive p53 expression was significantly associated with post-menopausal status, axillary lymph node metastases and Ag-NOR counts, whereas low level Rb expression was significantly associated with tumor size. Moreover, the combination of p53 and Rb expression was significantly associated with Ag-NOR counts, although there was no significant association between p53 and Rb expression. In the univariate study, p53 expression as well as age and axillary lymph node metastases were significantly associated with survival, whereas Rb expression was not. In the multivariate study, p53 and/or Rb expression did not provide independent prognostic information, although axillary lymph node metastases was an important factor affecting survival. Our findings suggest that p53 and/or Rb expression may reflect tumor proliferation of breast cancer, but the prognostic value of such assays is limited.     

Nm23 expression versus helix-pomatia lectin-binding in human breast-cancer metastases

The relationship between nm23 expression, HPA staining, and other clinicopathological variables, i.e., axillary (AX) and internal mammary lymph node (IMN) metastases was assessed, and their prognostic values were analyzed. Positive HPA staining was found to be significantly correlated with tumor size, and with AX and IMN metastases, whereas negative nm23 expression was significantly correlated with AX and IMN metastases. A univariate study revealed that overall survival was correlated significantly with tumor size, AX and IMN metastases, and HPA staining, whereas nm23 expression did not appear to be a significant prognostic factor. In a multivariate study, only AX and IMN metastases were significant prognostic factors. When AX and IMN metastases were excluded from the Cox model, however, only HPA staining had independent prognostic value for survival. In this study, therefore, it may be concluded that HPA staining was more closely correlated with AX and IMN metastases, and with survival than nm23 expression.     

A case with breast cancer under the nipple who underwent breast conserving treatment

Although breast conserving treatment (BCT) has become the standard therapy for early breast cancer, breast removal is still recommended for patients with a tumor beneath the nipple or with Paget’s disease. We have employed transposition of a latissimus dorsi myocutaneous (LD-MC) flap after wide local excision of a tumor with the nipple-areola complex. A new nipple-areola complex was reconstructed on the LD-MC flap after breast irradiation. Utilizing reconstructive techniques, BCT will likely become the treatment of choice for more patients with early breast cancer.     

Investigation of radiation safety management of nuclear medicine facilities in Japan; contamination of radioactivity in the draining-water system. A Working Group of Japanese Society of Nuclear Medicine for the Guidelines of Nuclear Medicine Therapy

Radiation safety management condition in Japanese nuclear medicine facilities were investigated by the questionnaire method. The first questionnaire was asked in all Japanese 1,401 Nuclear Medicine facilities. Answers from 624 institutes (44.5%) were received and analyzed. The radiation-safety management in nuclear medicine institutes was considered to be very well performed everyday. Opinion for the present legal control of nuclear medicine institutes was that the regulation in Japan was too strict for the clinical use of radionuclides. The current regulation is based on the assumption that 1% of all radioactivity used in nuclear medicine institutes contaminates into the draining-water system. The second questionnaire detailing the contamination of radioactivity in the draining-water system was sent to 128 institutes, and 64 answers were received. Of them, 42 institutes were considered to be enough to evaluate the contamination of radioactivity in the draining-water system. There was no difference between 624 institutes answered to the first questionnaire and 42 institutes, where the radioactivity in the draining-water system was measured, in the distribution of the institute size, draining-water system equipment and the radioactivity measuring method, and these 42 institutes seemed to be representative of Japanese nuclear medicine institutes. Contamination rate of radioactivity into the draining system was calculated by the value of radioactivity in the collecting tank divided by the amount of radionuclides used daily in each institute. The institutes were divided into two categories on the basis of nuclear medicine practice pattern; type A: in-vivo use only and type B: both in-vivo and in-vitro use. The contamination rate in 27 type A institutes did not exceed 0.01%, whereas in 15 type B institutes the contamination rate distributed widely from undetectable to above 1%. These results indicated that the present regulation for the draining-water system, which assumed that 1% of all radioactivity used in nuclear medicine institutes contaminated into draining-water system, should be reconsidered in nuclear medicine facilities where radionuclides are used only in in-vivo studies.     

Differential effects of lidocaine and mexiletine on relaxations to ATP-sensitive K+ channel openers in rat aortas

BACKGROUND: In cardiac myocytes, lidocaine reduces but mexiletine increases adenosine triphosphate (ATP)-sensitive K+ currents, suggesting that these class Ib antiarrhythmic drugs may differentially modify the activity of ATP-sensitive K+ channels. The effects of lidocaine and mexiletine on arterial relaxations induced by K+ channel openers have not been studied. Therefore, the current study was designed to evaluate whether lidocaine and mexiletine may produce changes in relaxations to the ATP-sensitive K+ channel openers cromakalim and pinacidil in isolated rat thoracic aortas. METHODS: Rings of rat thoracic aortas without endothelia were suspended for isometric force recording. Concentration-response curves were obtained in a cumulative fashion. During submaximal contractions to phenylephrine (3 x 10(-7) M), relaxations to cromakalim (10(-7) to 3 x 10(-5) M), pinacidil (10(-7) to 3 x 10(-5) M), or diltiazem (10(-7) to 3 x 10(-4) M) were obtained. Lidocaine (10(-5) to 3 x 10(-4) M), mexiletine (10(-5) to 10(-4) M) or glibenclamide (5 x 10(-6) M) was applied 15 min before addition of phenylephrine. RESULTS: During contractions to phenylephrine, cromakalim and pinacidil induced concentration-dependent relaxations. A selective ATP-sensitive K+ channel antagonist, glibenclamide (5 x 10(-6) M), abolished these relaxations, whereas it did not alter relaxations to a voltage-dependent Ca2+ channel inhibitor, diltiazem (10(-7) to 3 x 10(-4) M). Lidocaine (more than 10(-5) M) significantly reduced relaxations to cromakalim or pinacidil in a concentration-dependent fashion, whereas lidocaine (3 x 10(-4) M) did not affect relaxations to diltiazem. In contrast, mexiletine (more than 10(-5) M) significantly augmented relaxations to cromakalim or pinacidil. Glibenclamide (5 x 10(-6) M) abolished relaxations to cromakalim or pinacidil in arteries treated with mexiletine (10(-4) M). CONCLUSIONS: These results suggest that lidocaine impairs but mexiletine augments vasodilation mediated by ATP-sensitive K+ channels in smooth muscle cells.     

Highly sensitive analysis for N-myc amplification in neuroblastoma based on fluorescence in situ hybridization

BACKGROUND/PURPOSE: The N-myc amplification status in neuroblastoma has been evaluated previously for the whole tumor by the Southern blot method. The aim of this study is to evaluate the effectiveness of the fluorescence in situ hybridization (FISH) method to analyze N-myc amplification in neuroblastoma and compare the findings with those using the Southern blot method. METHODS: In 26 neuroblastoma primary tumors and metastatic lesions, the N-myc amplification status was evaluated by both the Southern blot method and FISH method. RESULTS: Of the 22 samples with no N-myc amplification using Southern blot, no cells with N-myc amplification using FISH were present in 21 of the samples. However, one metastatic liver lesion showed 16% of the nuclei to display more than 10 copies of N-myc based on FISH analysis. In the 4 remaining samples with N-myc amplification using the Southern blot method (17 copies, 15 copies, 6 copies, and 3 copies), the rates of cells with more than 10 copies of N-myc based on a FISH analysis were 79%, 68%, 94%, and 9%, respectively. CONCLUSIONS: The FISH method can detect more accurately N-myc amplification than the Southern blot method either when the rate of cells with N-myc amplification is low or intratumor heterogeneity is present.