Targeted chemotherapy in mice with peritoneally disseminated gastric cancer using monoclonal antibody-drug conjugate.

The murine monoclonal antibody A7 (MAb A7) is reactive against most human gastric cancer cell lines. Using a nude mouse peritoneal dissemination model of human gastric cancer, we investigated targeted chemotherapy using a conjugate of neocarzinostatin (NCS) with MAb A7 (A7-NCS). After demonstrating cytotoxicity of the complex against the human gastric cancer cell line MKN45 in vitro, we intraperitoneally injected A7-NCS, NCS or saline into nude mice bearing peritoneally disseminated human gastric cancer. A7-NCS inhibited peritoneal dissemination significantly more effectively than NCS. MAb A7 may prove to be an effective carrier for antineoplastic drugs in patients with peritoneal dissemination of gastric cancer.

Changes in expression of the antigen recognized by monoclonal antibody A7 in human pancreatic carcinoma cells following exposure to anticancer agents.

Techniques which can increase the expression level of tumor-associated antigens may improve immunotargeting therapy. We studied the reactivity of MAb A7 toward an antigen expressed on the surface of the human pancreatic cancer cell line HPC-YS after treatment with various antitumoral agents. When we applied 1 microg/ml mitomycin C (MMC) or 0.1 microg/ml neocarzinostatin (NCS) for 1 h, A7 recognizing antigen expression was enhanced until 24 h after the treatments. At a dose that completely suppressed cell growth, increased antigen expression was maintained for 96 h. Therefore, this study suggests that the combined application of an anticancer drug and MAb A7 may be useful for immunotargeting chemotherapy.

Reduced blood accumulation of biotinylated monoclonal antibody A7 after the subsequent administration of avidin.

Clear immunoscintigraphy with radiolabeled monoclonal antibodies (MAbs) requires a high tumor tissue/blood ratio of radioactivity. In this study, we attempted to obtain a high tumor tissue/blood ratio by the active removal of radiolabeled MAb from the circulation, using the avidin-biotin system. Biotinylated 125I-labeled MAb A7 was injected intravenously into nude mice bearing a human colon cancer (WiDr) xenograft. Avidin was injected 24 h later. The tumor tissue/blood ratio of radioactivity was almost four times that of controls. These results suggest that biotinylated 125I-labeled MAb A7 and avidin are potentially useful for the rapid immunodetection of human colon cancer.

The effect of intravenous and intra-tumoural chemotherapy using a monoclonal antibody-drug conjugate in a xenograft model of pancreatic cancer.

In order to investigate the efficacy of the intra-tumoural administration of an anticancer drug-monoclonal antibody conjugate in athymic nude mice bearing xenografts of a human pancreatic carcinoma, we examined the clearance of the murine monoclonal antibody A7 from the xenografts after intravenous or intra-tumoural administration and measured the antitumour effect of neocarzinostatin conjugated to MAb A7 following intravenous or intra-tumoural injection. Compared with 125I-labelled normal mouse IgG, a larger amount of 125I-labelled A7 remained in the tumour after both intravenous and intra-tumoural injection, and a significantly larger amount of 125I-labelled A7 remained in the tumour after intra-tumoural injection than that after intravenous injection. Moreover, a larger amount of 125I-labelled A7-NCS localized in the tumour after intra-tumoural injection than that after intravenous injection. Neocarzinostatin conjugated to MAb A7 showed greater activity against human pancreatic cancer than neocarzinostatin alone after both intravenous and intra-tumoural administration. Tumour growth was suppressed completely by the intra-tumoural administration of A7-NCS at a dose that did not suppress tumour growth via the intravenous route. These observations suggest that the intra-tumoural injection of neocarzinostatin conjugated to MAb A7 offers promise in treating pancreatic carcinoma.

[Bilateral pneumothorax and rupture of dissecting aortic aneurysm following a mitral valve replacement in Marfan syndrome: a case report].

A 12-year-old girl with Marfan syndrome was referred to our department because of severe mitral regurgitation. The patient was pale and in a pre-shock condition. The echocardiogram revealed a chordae rupture of the posteromedial papillary muscle at the posterior leaflet. A moderate dilatation of the aortic annulus (30 mm) and the ascending aorta (45 mm) without aortic regurgitation was also observed. Emergent mitral valve replacement was performed without replacement of the aortic valve or the ascending aorta. After two months, the patient developed bilateral pneumothorax, which was resistant to continuous suction therapy and finally required surgical treatments. Despite the relatively uneventful recovery, a lethal rupture of dissecting aortic aneurysm into the pericardial cavity, developed four months after the initial operation. We discussed the desired surgical approaches and respiratory problems in patients with connective tissue disorder.

[A case report: Rastelli operation for double outlet right ventricle associated with a chordae insertion to the infundibular septum].

A successful Rastelli operation for double outlet right ventricle (DORV) with a chordae insertion of the tricuspid valve to the infundibular septum was reported. A patient was a 6-year-old boy and the diagnosis was DORV, d-malposition of the aorta and pulmonary stenosis. The infundibular septum was not resected but mobilized with two incision, one anterior and vertical, and other one subaortic. This procedure allowed the construction of the tunnel similar to the closure of a large, subarterial VSD and of the straight unobstractive left outflow tract. The postoperative echocardiographic and angiographic examination revealed neither the pressure gradient in the left ventricular outflow tract nor the tricuspid regurgitation.

[Diagnosis and targeting therapy of colorectal cancer using antibody].

Application of monoclonal antibody for diagnosis and therapy of colorectal cancer was reviewed. The history and present status of radioimmunoimaging of cancer were presented. Immuno-guided surgery using radiolabeled antibody and hand-aided detector during surgery is a promising approach for complete excision of cancerous region. Also, a new antibody labeling method using a micro-magnet instead of radioisotopes may lead to a new era in immunodiagnosis of cancer. Finally, immunotargeting chemotherapy using antibody-anticancer drug conjugates was reviewed. A new type of immunoconjugate composed of human/mouse chimeric antibody and Neocarzinostatin seems to be one of the most promising drug formulas for immunotargeting chemotherapy.

[Influence of the surgical stress on graft-versus-host reaction in mice--the suppressor activity to GVH reaction induced by the surgical operation].

The influences of surgical stress on the local graft-versus-host reaction (GvHR) in F1 mice were studied. Skin incision to the F1 mice inflicted to 1 day prior to injection of parental spleen cells produced impairment of popliteal lymph node enlargement, however, this effect was not observed when GvHR was induced on the day 3 and 5 after operation. Spleen cells from hind limb amputated F1 mice exhibited suppressor activity on local GvH reaction when injected into naive syngenic F1 mice together. This suppressor activity was markedly detected 3 and 24 hrs after operation, then gradually reduced and completely disappeared on the day 7. These suppressor activity completely disappeared by the treatment with anti-Thy 1.2. antibody and complement. These results showed that GvH reaction was suppressed by the surgical stress, and this suppression was due to suppressor T lymphocytes in the spleen.

Evaluation of neuropsychological function in patients with liver cirrhosis with special reference to their driving ability.

Ability to drive an automobile was evaluated in 16 patients with well compensated liver cirrhosis. Four tests were performed, namely the emergency reaction test, the continuous emergency reaction test, the signal confirmation test and the accelerator reaction test. Test scores were compared to those of a group of age-matched healthy volunteers. 31% of patients were found to be unfit to drive. Alcoholic cirrhotics fared as poorly as non-alcoholic cirrhotics. In patients with subclinical hepatic encephalopathy (defined by neuropsychologic testing), 44% were unfit to drive. Routine testing of cirrhotic patients for ability to drive could have a major impact on motor vehicle accident rates.

On the histochemistry of neuronal and glial TPPase.

Thiamine pyrophosphatase (TPPase) activity in the neuronal Golgi apparatus and in glia cells was investigated using different tissue preparation techniques such as fresh frozen and fixed frozen sections, the effect of ATP as an activator and the fine structural localization of the enzyme activity in glia cell processes and the glio-vascular contact point was observed under the electron microscope. TPPase was demonstrated in the glia cell using only a fixed frozen tissue preparation, activated by ATP with the concentration as low as 0.1 mM. On the contrary, enzyme activity was detected in the Golgi apparatus of the neuron regardless of tissue preparation, fresh or fixed frozen, or by the addition of ATP to the incubation medium. The identity of the glia cell type showing a positive reaction for TPPase is thought to be part astrocyte and part microglia. The fine structural observation of TPPase localization at the glia cell process ending, particularly at the glio-vascular and glio-fibral (medullated fiber) contact point suggests a functional role of this enzyme for transport of metabolites through glia cell processes to neurons and/or the blood capillary, and also may play a role in regulation of co-carboxylase.

Fine structural correlations between the muscle pathology of amyotrophic lateral sclerosis (ALS) patients and experimental Ca-Mg deficient rats.

Nonspecific myofibrillar changes such as streaming of the Z-line, formation of rod-like structures, satellitosis, proliferation of sarcolemmal nuclei and papillary projection of the sarcolemma were recognized as a disorganization of the muscle itself. In addition, fine structural pathology in ALS specimens showed characteristically a pig-tail formation - 'Zopfformation' - which has been considered to have a neurogenic origin.

[A case report of ultrasonic decalcification for calcified bicuspid aortic valve stenosis].

Calcified stenotic aortic valve are usually replaced by the artificial valve. Recently, we are trying to decalcify the calcification of the aortic valve by ultrasonic energy. This case report reviewed a small physique, 47 year old female, who had the calcified stenotic bicuspid aortic valve and the narrow aortic valve ring (18 mm diameter). The ultrasonic decalcification was performed by the ultrasonic surgical instrument, SUS-201D (ALOKA), ultrasonic energy output range from 25% to 45%. After the ultrasonic decalcification, the movability of the aortic cusps were improved and the aortic opening was enlarged. The systolic pressure gradient across the aortic valve was improved from 100 mmHg to 20 mmHg. This method "ultrasonic decalcification" is very useful technique for the repair of the calcified aortic valve stenosis.

[Adenosine triphosphate: an additive myocardial agent in cardioplegia].

The potential for enhancing myocardial protection by adding ATP to cardioplegic solution was investigated in a rat heart model of cardiopulmonary bypass. Concentration of added ATP was 0.01 mM and 0.1 mM. The hearts were perfused by working heart model for 10 min as a baseline hemodynamic variables and they underwent infusion of cardioplegic solution followed by 20 min cardiac arrest with myocardial temperature at 37 degrees C. The ATP added group revealed good % recovery of aortic flow after 20 min period of ischemic arrest compared to the ATP free control group. But there is no difference between 0.01 mM and 0.1 mM ATP added group. ATP added group showed higher ATP concentration in myocardium and ATP/ADP and energy charge after 20 min of ischemic arrest than ATP free control group. But total adenosine showed no difference. The result suggested that ATP was a protective myocardial agent in cardioplegic solutions and ATP might protect membrane-bound ATP dependent process by ensuring energy supply.

Expression of matrix metalloproteinase-13 and tissue inhibitor of metalloproteinase-1 in acute liver injury.

BACKGROUND/AIMS: Matrix metalloproteinase-13, one of the principal neutral proteinases capable of cleaving native fibrillar collagens, is important in the degradation and remodeling of extracellular matrix. However, its precise expression in liver injury has not been characterized. We examined the kinetics of the expression of matrix metalloproteinase-13 and one of its specific inhibitors, tissue inhibitor of metalloproteinase-1, in acute liver injury in rats. METHODS: Acute liver injury was induced by administration of carbon tetrachloride or two different doses of D-galactosamine hydrochloride in Wistar rats. Hepatic matrix metalloproteinase-13 and tissue inhibitor of metalloproteinase-1 mRNA levels were then examined by Northern blotting. RESULTS: All rats survived after liver injury induced by carbon tetrachloride or low doses of D-galactosamine hydrochloride. However, rats died 5 days after induction of liver injury by high doses of D-galactosamine hydrochloride. In carbon tetrachloride-induced liver injury, matrix metalloproteinase-13 mRNA was transiently increased between 6 h and 1 day after injury. Tissue inhibitor of metalloproteinase-1 mRNA expression was increased between 6 h and 3 days after the peak of matrix metalloproteinase-13 expression. Similar patterns of matrix metalloproteinase-13 and tissue inhibitor of metalloproteinase-1 expression were observed in low-dose D-galactosamine hydrochloride-induced liver injury. In contrast, in high-dose D-galactosamine hydrochloride-induced liver injury, tissue inhibitor of metalloproteinase-1 expression peaked before matrix metalloproteinase-13 expression, which was increased 2 days after injury. Both mRNA levels continued to increase until death. CONCLUSIONS: Transient expression of matrix metalloproteinase-13, followed by that of tissue inhibitor of metalloproteinase-1, was observed during recovery from acute liver injury induced by carbon tetrachloride and low-dose D-galactosamine hydrochloride. In contrast, disordered expression of matrix metalloproteinase-13 was observed in fatal liver injury caused by high-dose D-galactosamine hydrochloride. These results indicate that matrix metalloproteinase13 plays an important role in the early phase of recovery from liver injury.

Spatial distribution of tissue inhibitor of metalloproteinase-1 mRNA in chronic liver disease.

BACKGROUND/AIMS: Tissue inhibitor of metalloproteinase-1, a specific inhibitor of matrix metalloproteinases, plays an important role in the pathogenesis of fibrosis and tumor progression. However, the precise expression of tissue inhibitor of metalloproteinase-1 messenger RNA in human hepatic fibrosis has not yet been defined. We investigated the spatial distribution of tissue inhibitor of metalloproteinase-1 messenger RNA in chronic human liver disease. METHODS: Northern and in situ hybridization of probes to tissue inhibitor of metalloproteinase-1 messenger RNA were performed in specimens from 16 surgically resected human livers. Immunohistochemical staining of sections for tissue inhibitor of metalloproteinase-1 and immunoelectron microscopy were also performed. RESULTS: Northern hybridization demonstrated that expression of tissue inhibitor of metalloproteinase-1 messenger RNA was increased 3.9-fold in mild chronic hepatitis, 6.8-fold in moderate chronic hepatitis, and 6.4-fold in cirrhosis, compared with control liver. In situ hybridization showed the expression of tissue inhibitor of metalloproteinase-1 messenger RNA in spindle-shaped cells in the fibrous septa and lobules in chronic hepatitis and cirrhosis; these cells were immunohistochemically positive for a-smooth muscle actin. Immunoelectron microscopy revealed localization of tissue inhibitor of metalloproteinase-1 in between fibers, to the rough endoplasmic reticula of stellate cells located in the lobules and periportal areas, and to fibroblasts in the fibrous septa. These results indicate that tissue inhibitor of metalloproteinase-1 was produced mainly by stellate cells in the specimens of chronic liver diseases. CONCLUSIONS: Expression of tissue inhibitor of metalloproteinase-1 messenger RNA is increased in hepatic fibrosis and stellate cells are involved primarily in its expression.

[Effect of controlled freezing-point storage of hearts--combined effect of controlled freezing-point storage and verapamil].

Controlled Freezing-point Storage (CF Storage) is a new method of preserving foods in minus non-frozen temperature range. Therefore, the authors tried to apply this method to organ preservation. We investigated the effect of controlled freezing-point storage of hearts on ventricular function in isolated perfused rat heart (male, Sprague dawley strain, body weight about 300 g), and the combined effect of controlled freezing-point storage and Verapamil (0.5 mg/l). The hearts were perfused by working heart mode for 10 min, and received infusion of cardioplegic solution which was followed by 4 hours of cardiac arrest. The hearts were stored for 4 hours in Euro-Collins solution at 4 degrees C (4 degrees C group) and minus non-frozen temperature (CF storage group), and were stored for 4 hours in Euro-Collins solution containing Verapamil at minus non-frozen temperature (CF-Ve storage group). Then, the aerobic reperfusion by working heart mode was continued for an additional 30 min. The recovery rate of cardiac output was 33.5%, 62.5% and 74.2% respectively of the preischemic value in 4 degrees C group, CF storage group and CF-Ve storage group. The recovery rate of cardiac output in 4 degrees C group was significantly lower than that in CF storage group and CF-Ve storage group (p less than 0.01), and significant difference was noted between CF storage group and CF-Ve storage group (p less than 0.05). ATP activity after 4 hours of cardiac arrest was 13.9 micrograms/mg, 18.7 micrograms/mg and 18.3 micrograms/mg in 4 degrees C group, CF storage group and CF-Ve storage group respectively. ATP activity after 4 hours of cardiac arrest in 4 degrees C group was significantly lower than that in CF storage group and CF-Ve storage group (p less than 0.05). These data suggest that CF storage of hearts has a protective effect against an ischemic insult upon myocardial cell during hypothermic cardiac arrest, and the best metabolic and functional protection appeared when the hearts were stored in Euro-collins solution containing Verapamil at minus non-frozen temperature.