A randomized controlled study evaluating the efficacy of aprepitant for highly/moderately emetogenic chemotherapies in hematological malignancies.

Chemotherapy-induced nausea and vomiting (CINV) is a serious complication of treatments of hematological malignancies. Although aprepitant, an NK1 receptor antagonist, has been shown to control CINV in highly emetogenic therapies for solid tumors, the antiemetic effect of this agent in hematological chemotherapies is not well established. In this randomized controlled trial, we examined the additional effect of aprepitant in combination with conventional 5HT3 blocker-based prophylaxis for CINV in highly or moderately emetic chemotherapies for hematological malignancies (n = 41). The complete response rate, defined as no emetic episodes and no salvage treatments, was significantly higher in the aprepitant arm than the control arm (82 versus 47 %, p = 0.026), with no increase in severe adverse effects. However, the difference of nausea, measured with visual analog scale, and of oral intake impairment was moderate, which suggests insufficiency of blocking NK receptor for these events. Furthermore, sub-group analysis revealed that merit of aprepitant addition depends on treatment regimens. Our results indicate the overall advantage of applying aprepitant in the control of CINV in hematological malignancies and the need for further refinement of anti-CINV strategies, including stratification according to regimen.

Analysis of cell kinetics after gamma-ray irradiation using an anti-BrdU monoclonal antibody.

The cell cycle was analyzed using double staining with an anti-bromodeoxyuridine (BrdU) monoclonal antibody and propidium iodide (PI). Changes in cell kinetics after irradiation were compared with those seen by the conventional PI-based DNA histogram method. The effect of irradiation on cell kinetics has been studied primarily by counting G2-arrested cells. By the present BrdU method, a rapid transition from G1 to S-phase was observed within 2 h of irradiation, followed by G1 block. Cells in the S-phase progressed to G2+M where they arrested, resulting in a decreased percentage of S cells (<5%). Release of G1 block occurred after 8 h, and G2+M cells returned to G1 after >18 h. The initial G1 arrest induced by irradiation was confirmed for the first time by the present BrdU-PI double staining.

Cell kinetic changes in cultured tumor cells after treatment with radiation and chemotherapy.

Flow cytometry instantaneously determines the percentages of cells in various cell cycle phases to rapidly evaluate effects of irradiation and anti-cancer drugs. We studied these using growth curve analysis and 5-bromodeoxyuridine propidium iodine (BrdU)-(PI) double staining. With conventional DNA histogram methods, determination of S phase fraction was difficult because of overlapping DNA content between G1 and early S phases and between late S and G2 phases. Double staining directly differentiated G1, S, and G2 + M phases. By double staining, rapid transition from G1 to S occurred within 4 h after irradiation or after the drug treatments, and initial G1 arrest induced by irradiation was confirmed for the first time.

Enhancement of thermal damage in murine tumors by hydralazine-induced modification of blood flow and oxygen tension.

We investigated changes in blood flow in normal muscle and in SCC-VII tumors treated by hyperthermia combined with hydralazine, to evaluate the enhancement of thermal tumor damage by hydralazine. We studied SCC-VII tumor-bearing C3H/He mice. Hydralazine was administered by intraperitoneal injection, and tumors were heated by a water bath. We measured blood flow using the laser Doppler method, and oxygen tension using polarography. The response of tumors to therapy was assessed using a growth delay analysis. In tumors, blood flow and O-2 tension significantly decreased with increasing doses of hydralazine. Compared to tumors treated by hydralazine alone or by hyperthermia alone, tumor blood flow was significantly decreased in the group treated by hyperthermia with hydralazine. In tumors treated by hyperthermia with hydralazine, blood flow was significantly decreased with increasing Hyd doses, heat durations, and temperatures. In normal muscle, no decrease in blood flow was induced by hyperthermia, hydralazine, or their combination. In tumors treated by hyperthermia (43 degrees C, 20 min) with hydralazine, a maximum additional growth delay was observed. Our results suggest that a decrease in tumor blood flow caused by hydralazine plays an important role in enhancement of the hyperthermic antitumor effect.

Quantitative studies on translocation and metabolic conversion of lysophosphatidylcholine incorporated into the membrane of intact human erythrocytes from the medium.

The fate of palmitoyl-lysophosphatidylcholine (lysoPC) incorporated into the membrane of intact human erythrocytes from a medium was investigated under nonhemolytic conditions at 37 degrees C by means of 14C-labeled tracers. The lysoPC was first incorporated into the outer half of the membrane lipid bilayer and then gradually translocated into the inner half during the incubation. At the same time it was metabolically converted into phosphatidylcholine (PC) and free fatty acid (FFA) plus glycerophosphorylcholine by the actions of acyltransferase and lysophospholipase, respectively. The half times of the conversion were about 14 h, while the value of 0.5 h was obtained when the half time was measured with the hemolysate of the lysoPC-loaded erythrocytes. Chymotrypsin treatment of unsealed ghosts caused a definite decrease in lysophospholipase activity, while similar treatment of resealed ghosts did not. This together with other evidence already reported in the literature suggests that both lysophospholipase and acyltransferase may be located in the inner surface of the membrane. The above findings strongly suggest that the most of the lysoPC loaded to the membrane is gradually translocated from the outer to the inner half of the bilayer and soon converted to either PC or FFA.

Immunohistochemical analysis of intercellular adhesion molecule-1 expression in human gastric adenoma and adenocarcinoma.

In this study, we examined the distribution of intercellular adhesion molecule-1 (ICAM-1) in gastric adenomas and carcinomas immunohistochemically at the light and electron microscopic levels. ICAM-1 was expressed on tumour cells in 12 of 28 gastric carcinomas and in 3 of 11 adenomas but not on most normal gastric epithelial cells. ICAM-1 was localized on luminal sites of neoplastic glands in adenomas and in intestinal-type carcinomas, and rarely on the surface of tumour cells of diffuse carcinomas. Expression of ICAM-1 on the tumour cells was more frequent in intestinal-type than diffuse carcinomas (P < 0.005). At the ultrastructural level, ICAM-1 was present prominently on the apical membrane and weakly on the lateral surface of the tumour cells of the intestinal-type carcinoma and also localized on the perinuclear membrane and the membrane of the endoplasmic reticulum of cancer cells. There was no significant association between ICAM-1 expression and HLA antigen expression or the number of infiltrating lymphocyte subsets. These results may implicate the synthesis of ICAM-1 by gastric cancer cells, but the expression is infrequent and may not be sufficient for host immune surveillance of the tumour cell.

Analysis of cell kinetics after irradiation by flow cytometry: proliferative ability of G2-blocked cells after cell sorting.

Irradiation, dose-dependently, increases the percentage of cells in the G2 + M phase and the duration of cell cycle arrest, reflected by changes in cell kinetics in the first 48 h. Cells in the G2 + M phase are considered radiosensitive but little is known about their proliferative ability. We studied proliferative ability of irradiated G2 phase-arrested cells following flow cytometric cell sorting using fluorescent Hoechst 33342 staining. Although proliferative ability of non-irradiated cells was not different between G1 and G2 + M phases, cells arrested in G2 + M after irradiation, especially at higher doses, showed less ability than cells in G1. Proliferative ability also correlated well with of G2 block duration.

Enhancement of antitumor effect of hyperthermia with glucose administration in murine mammary carcinoma.

Antitumor effects of hyperthermia are enhanced by lowering the pH in the tumor tissue with administration of glucose. This decreased pH in the tumor tissue with glucose administration was determined using mouse experimental tumors. 31P-MRS microelectrodes were used for the measurement of pH. By using these two measurement methods, time course change in the tumor tissue was determined in the controls and the groups treated with 6 g/kg of intraperitoneal glucose. The determination of pH with 31P-MRS was calculated from the chemical shift of the peak of creatine phosphate (Pcr) and that of inorganic phosphate (Pi). Following glucose administration, the tumor tissue showed a decrease of 0.3 pH units with the microelectrode method, but did not show any significant decrease in pH with the MRS determination. This finding suggested that 31P-MRS showed intracellular pH (pHi) due to the localization of Pi and that the microelectrode indicated interstitial or extracellular pH (pHe). The ATP/Pi ratio obtained in tumor tissue 24 h after heat treatment (with, without glucose) was correlated with tumor inhibition.

Studies on hyperthermia combined with arterial blockade for treatment of tumors: (Part I) effectiveness of hyperthermia combined with arterial ligation.

Arterial ligation was combined with hyperthermia in rabbits with VX2 tumors implanted in the leg. For seven days after arterial ligation, blood flow was decreased and the pH was low in both normal muscle and tumor tissue. The temperature of normal muscle and tumor tissue increased faster and reached a higher level on heating immediately after ligation than without ligation. The antitumor effect of hyperthermia was stronger immediately after ligation than two or seven days afterwards. However, damage to normal muscle was severe with this combination therapy, so a better method of therapeutic arterial blockade is needed.

Studies on hyperthermia combined with arterial therapeutic blockade for treatment of tumors: (Part III) effectiveness of hyperthermia combined with arterial chemoembolization using degradable starch microspheres on advanced liver cancer.

Arterial chemoembolization using degradable starch microspheres and adriamycin was combined with hyperthermia to treat advanced liver cancer. The prolonged peak adriamycin level in hepatic venous blood suggested that the drug persisted for longer in the liver after injection containing microspheres. Heating efficiency was increased more in tumor tissue than in normal liver tissue after embolization. This combined therapy was performed in eight patients with advanced liver cancer and was effective in three (complete or partial remission). The mean survival time was 25 weeks and there were no severe side effects. This combined therapy may be useful for liver cancer.

Studies on hyperthermia combined with arterial blockade for treatment of tumors: (Part II) effectiveness of hyperthermia combined with arterial embolization using degradable starch microspheres.

The efficacy of temporary arterial embolization using degradable starch microspheres combined with hyperthermia was investigated in rabbits bearing VX2 tumors. Microsphere injection caused a marked decrease of tumor blood flow and pH. During heating, there was a marked increase of the maximum temperature in tumor tissue compared with normal muscle. Tumor growth was suppressed 330% times at 3 weeks after hyperthermia alone and 270% times following combined treatment with microspheres and hyperthermia. Damage to normal muscle tissue was mild. In conclusion, this combination therapy may be useful for causing selective tumor damage and reducing the effect on normal tissues.

In-111 labeled leukocyte scintigraphy in patients with suspected inflammation after failed antibiotic therapy.

In-111 labeled leukocyte scintigraphy (In-111 WBC scan) was performed in 16 patients with inflammation suspected on the basis of laboratory findings, symptoms, and diagnostic imaging, but who had failed antibiotic therapy. In-111 WBC scans revealed an abnormal focus of radiotracer activity (positive scans) in five of 16 patients. No correlation was found between the peripheral WBC count and accumulation of In-111 WBC. Inflammatory disease suspected on the basis of the CRP level should be considered when In-111 WBC scanning results in negative findings. Our results indicated that In-111 WBC scanning has low sensitivity after antibiotic therapy. Selection of patients on the basis of persistent elevation of CRP may be valuable.

Effects of right hemithoracic irradiation on ceftazidime penetration into the alveolar space in rats.

The effect of thoracic irradiation on antibiotic penetration into the alveolar space was determined in a hemithoracic irradiation rat model to evaluate radiation-induced acute alveolar injury at various time intervals. The results of this investigation may be summarized as follows: (1) The transfer of ceftazidime (CAZ: Modacin) from blood to lung tissue, that is, the permeability of pulmonary capillary epithelium, and the transfer of ceftazidime from lung tissue to bronchoalveolar lavage fluid, that is, the permeability of the alveolar epithelium, peaked at 4 to 5 weeks after thoracic irradiation; (2) the time course of change in the absolute concentration of ceftazidime in lung tissue showed a significant increase not only in the irradiated lung but also in the contralateral non-irradiated lung 3 days or more after irradiation. The finding that the administration of antibiotics may cause a significant increase in drug concentration in lung tissue even in the contralateral lung at 3 days after irradiation suggests that the change induced at the alveolar level immediately after irradiation affects the non-irradiated lung field through an as yet unknown mechanism.

New technique for mediastinal temperature measurement in hyperthermic cancer treatment: balloon catheter in the azygos vein.

The temperature in the mediastinum during hyperthermia is difficult to determine accurately. We measured the temperature in the azygos vein, using a new technique, and compared the measurements with temperatures in the oesophagus. Eight patients with mediastinal tumours resulting from lung cancer or oesophageal cancer were given hyperthermo-radiotherapy. The temperatures in the azygos vein and in the oesophagus were measured before and during blockage of the blood flow of the azygos vein using an angiographic balloon catheter. None of the patients had complications as a result of these procedures, and hyperthermia by capacitative heating was safely performed. The temperature in the azygos vein increased by a mean of 1.7 degrees C (0.2-2.8 degrees C) after blockage of the blood flow. The temperature in the oesophagus was 0.83 +/- 1.09 degrees C (mean +/- SD) higher than that in the azygos vein. Measurement of the temperature in the azygos vein gives a more accurate estimate of mediastinal temperature than does oesophageal temperature but it is an invasive procedure.

Usefulness of dual SPECT with Tc-99m pyrophosphate and Tl-201 to predict further events after acute myocardial infarction with single-vessel coronary artery disease.

PURPOSE: This study was undertaken to determine whether the findings of dual SPECT with Tc-99m pyrophosphate (PYP) and Tl-201 were predictive of further cardiac events after acute myocardial infarction. METHODS: The authors evaluated 88 patients with acute myocardial infarction who underwent dual SPECT for single-vessel coronary artery disease. RESULTS: Twenty-nine patients showed overlapping of Tc-99m PYP and Tl-201 in the same location (overlap-positive group), and 59 patients had no overlap (overlap-negative group). In patients in the overlap-positive group, the incidence of subsequent events was significantly higher than in patients in the overlap-negative group (P < 0.001). In the overlap-positive group, the number of overlap segments in patients with further events was significantly greater than that in patients without further events (P < 0.005). CONCLUSIONS: Areas with overlapping of Tc-99m PYP and Tl-201 may contain jeopardized myocardium. These results suggest that patients who have a Tc-99m PYP and Tl-201 overlap-negative scan are a low risk group, whereas patients who have more overlapping segments may require catheterization and revascularization. Thus simultaneous SPECT imaging with Tc-99m PYP and Tl-201 might be useful to identify patients with greater ischemic risk after acute myocardial infarction.

[Analysis of intermittent blood flow in experimental tumor using a transparent chamber].

Using a transparent chamber under a microscopic recording system equipped with a CCD camera, we measured real-time fluctuations of microvessel blood flow in an experimental murine tumor. Seven microvessels of the tumor showed 3-4 fluctuations in blood flow (intermittent blood flow) during the observation period (2 hours). Within the 120 min. monitoring period, we obtained approximately a maximum 52.5% of decreased blood flow. The duration of change in blood flow ranged from 15-45 min. Similar temporal fluctuations were seen in experimental animal tumors. Our data clearly demonstrated that fluctuations in blood flow (intermittent blood flow) in tumors are a common feature.

[Analysis of tumor vascular density using the window chamber technique].

Vascular structure is indispensable for a tumor to maintain its growth. The structure also affects drug delivery over chemotherapy, and oxygen tension in the tissue during irradiation. Therefore, measuring the vascular density in the tumor will help to reach an expected level or to evaluate the effect of those therapies. Through previous observation, changes in vascular structure, like the vascular density, have been reported. However, the tissue was usually removed from the deceased animal and examined in the microscope. Not many studies have shown changes in one and the same animal's tissue. In this study, we used the dorsal flap window chamber technique, and observed changes in the development of vascular structure and vascular density in a tumor. Window chambers were attached to the backs of 3 rats, and they anesthetized every 24 hours for microscopic observation. Vascular growth in the tumor started 8 to 9 days after implantation. Their average vascular density was about 25%, and this ratio continued till necrosis started in the tumor. "Necrosis occurs in a tumor because its growth is so rapid that the vascular development can not maintain the same speed." This has been a kind of established theory. During observation, we recognized blood stagnation in vessels as the tumor grew, and the relationship between tumor size and its vascular density was significant (r = 0.926). Our data lead us to another conclusion that "Vascular growth is as fast as tumor growth, keeping a ratio of about 25%, but necrosis is caused by deficiency of blood flow circulation."

[Effect on hepatic function of hepatic artery infusion chemotherapy using epirubicin and mitoxantrone for advanced hepatocellular carcinoma].

This study was designed to assess the effect of hepatic dysfunction from hepatic artery infusion (HAI) in advanced hepatocellular carcinoma (HCC). The patients were randomly assigned to receive epirubicin (n = 12, Epi group) or mitoxantrone (n = 14, Mito group) once every 4 weeks between 1992 and 1996. HCC patients were given 6-8 mg of mitoxantrone or 30-40 mg epirubicin. There was hepatic dysfunction in 27 patients after HAI, showing similarly elevated GOT, GPT, and total bilirubin. In the Epi group, the GOT value was slightly higher than in the Mito group, but it was not significant. After HAI chemotherapy, the GOT value showed a more than two-fold elevation. Six patients in the Epi group and 2 in the Mito group showed a significant difference. Our results indicated that mitoxantrone had less impact on hepatic function following HAI therapy.

Evi1 defines leukemia-initiating capacity and tyrosine kinase inhibitor resistance in chronic myeloid leukemia.

Relapse of chronic myeloid leukemia (CML) is triggered by stem cells with a reconstituting capacity similar to that of hematopoietic stem cells (HSCs) and CML stem cells are a source of resistance in drug therapy with tyrosine kinase inhibitors (TKIs). Ecotropic viral integration site 1 (EVI1), a key transcription factor in HSC regulation, is known to predict poor outcomes in myeloid malignancies, however, incapability of prospective isolation of EVI1-high leukemic cells precludes the functional evaluation of intraindividual EVI1-high cells. Introduction of CML into Evi1-internal ribosomal entry site (IRES)-green fluorescent protein (GFP) knock-in mice, a versatile HSC-reporter strain, enables us to separate Evi1-high CML cells from the individual. Evi1-IRES-GFP allele models of CML in chronic phase (CML-CP), by retroviral overexpression of BCR-ABL and by crossing BCR-ABL transgenic mice, revealed that Evi1 is predominantly enriched in the stem cell fraction and associated with an enhanced proliferative as well as a leukemia-initiating capacity and that Evi1-high CML-CP cells exhibit resistance to TKIs. Overexpressing BCR-ABL and NUP98-HOXA9 in Evi1-IRES-GFP knock-in mice to model CML in blast crisis (CML-BC), in which Evi1-high cells turned to be a major population as opposed to a minor population in CML-CP models, showed that Evi1-high CML-BC cells have a greater potential to recapitulate the disease and appear resistant to TKIs. Furthermore, given that Evi1 heterozygosity ameliorates CML-CP and CML-BC development and that the combination of Evi1 and BCR-ABL causes acute myeloid leukemia resembling CML-BC, Evi1 could regulate CML development as a potent driver. In addition, in human CML-CP cases, we show that EVI1 is highly expressed in stem cell-enriched CD34+CD38-CD90+ fraction at single-cell level. This is the first report to clarify directly that Evi1-high leukemic cells themselves possess the superior potential to Evi1-low cells in oncogenic self-renewal, which highlights the role of Evi1 as a valuable and a functional marker of CML stem cells.

The pleiotrophin-ALK axis is required for tumorigenicity of glioblastoma stem cells.

Increasing evidence suggests that brain tumors arise from the transformation of neural stem/precursor/progenitor cells. Much current research on human brain tumors is focused on the stem-like properties of glioblastoma. Here we show that anaplastic lymphoma kinase (ALK) and its ligand pleiotrophin are required for the self-renewal and tumorigenicity of glioblastoma stem cells (GSCs). Furthermore, we demonstrate that pleiotrophin is transactivated directly by SOX2, a transcription factor essential for the maintenance of both neural stem cells and GSCs. We speculate that the pleiotrophin-ALK axis may be a promising target for the therapy of glioblastoma.