A novel splicing mutation in the ceruloplasmin gene responsible for hereditary ceruloplasmin deficiency with hemosiderosis.

Hereditary ceruloplasmin deficiency with hemosiderosis (aceruloplasminemia) is a newly recognized autosomal recessive disorder of copper-iron metabolism due to mutations in the ceruloplasmin (Cp) gene. We report here a novel mutation in the Cp gene in a 54-year-old Japanese woman with this disease. She showed clinical triad; diabetes mellitus, retinal degeneration and neurological disorder in her middle age. Laboratory findings were characteristic for no detectable serum ceruloplasmin and increased serum ferritin. Liver biopsy revealed excessive storage of iron in hepatocytes and magnetic resonance imaging of the brain was indicative of increased iron content in the basal ganglia, thalamus and dentate nucleus. The a-->g substitution at the splice acceptor site of the intron 6 (1209-2) caused a 8-bp deletion in Cp mRNA by defective splicing, resulting in a premature termination codon at the amino acid position 388. Truncation of Cp, even if effectively translated, may cause loss of its normal function because of drastic change in its triangular structure.

Telomerase, p53 and PCNA activity in osteosarcoma.

AIMS: The aim of this study was to investigate telomerase activity and to assess the correlation between telomerase activity, tumor proliferative activity and p53 overexpression in osteosarcoma tumor samples. METHODS: Using a telomerase polymerase chain reaction-enzyme-linked immunoassay based on the telomeric repeat amplification protocol method, we examined telomerase activity in 26 primary osteosarcoma specimens. P53 overexpression was identified using immunohistochemical staining, and tumor proliferative activity was assessed by immunohistochemical staining of PCNA. RESULTS: Telomerase activity was detected at a relatively low level in five of 26 osteosarcoma tissue specimens. P53 was detected in eight of 21 cases. There was no significant correlation between telomerase activity and p53 overexpression (p=0.325). There was a significant correlation between PCNA staining and telomerase activity (p=0.0075). CONCLUSION: Difference between the telomerase activity and p53 overexpression in osteosarcoma suggests that p53 and telomerase may not cooperate in tumor proliferation. Correlation of telomerase activity to PCNA expression suggests that telomerase activity may also useful for evaluate proliferative activity in osteosarcoma.

Reduction of VX2 transplanted tumor by granulocyte depletion using extracorporeal circulation on rabbit models.

To investigate the role of granulocytes and the effect of granulocytosis on malignancies, we studied the relationship between white blood cell and granulocyte (Gr) counts, and tumor size. As we have reported that cancer patients with high granulocyte (G/L) ratio have a poor clinical prognosis, normalization of the G/L ratio could contribute to improvement in their condition. Tumor bearing rabbits treated with the extracorporeal granulocytte depletion device showed a decrease in tumor size and no tumor necrosis. Tumor-bearing rabbits with no extracorporeal granulocyte depletion showed a remarkable tumor progression and an increase in Gr count. Depletion of granulocytes considerably improved the survival ratio of tumor-bearing animals. The device used in this study could be a potential treatment for malignant tumors.

Does extensive dissection of lymph nodes improve the results of surgical treatment of gastric cancer?

The results of surgical treatment of patients with advanced gastric cancer were analyzed retrospectively in terms of the extent of dissection of lymph nodes. The postoperative 5-year survival rate was 50.3% for 160 patients who had undergone potentially curative surgery with dissection of group 1, 2, and 3 lymph nodes (R3), compared with 48.8% for 185 patients with dissection of group 1 and 2 lymph nodes (R2). There was no difference in terms of survival between the two groups of patients. Of 160 patients receiving R3, 19 (11.9%) were found to be positive for metastasis to group 3 lymph nodes, and only 5 survived for more than 5 years after surgery (postoperative 5-year survival rate of 26.3%). In four of these five patients, metastases in the group 3 lymph nodes were confined to nodes in the hepatoduodenal ligament. Thus, it appears that in dissection of the deepest nodes, lymph nodes in the hepatoduodenal ligament are the most important to remove in the surgical treatment of advanced gastric cancer.

Cellular fibronectin in plasma: its implications in fibrinogen-associated cryoprecipitation and other related reactions.

Elimination of cryoprecipitable plasma components (cryogels) by cryofiltration from the circulating blood of patients with drug-resistant rheumatoid arthritis (RA) alleviates clinical symptoms including morning stiffness and arthralgia. The cryogels thus isolated from the blood were found to consist mainly of fibrinogen (Fbg) and fibronectin (FN). Analysis by immunoblotting with an anti-cellular FN monoclonal antibody revealed that cellular FN (cFN) co-existed with plasma FN (pFN) in the cryogels derived from the patients. Using an ELISA, we assessed cFN together with the total FN (pFN+cFN) in plasmas and cryogels derived from the patients. The cFN/total FN ratio was distinctly higher in the cryogels than in the plasmas, suggesting that cFN was more readily precipitated than pFN in association with Fbg under cold conditions.

Climatic adaptation in thermogenesis and thermal insulation in wood mice (Apodemus argenteus).

Wood mice (Apodemus argenteus) were trapped live at three different altitudes (below 1,000, 1,900, and 2,400 m) during a 1-year period (Feb. 1984-Jan. 1985). After remaining at the trapped locations for 10-14 days, they were transferred into a climatic chamber at an altitude of 610 m. Oxygen consumption (VO2) and colonic temperature (Tco) were measured at chamber temperatures (Ta) of five steps (30, 20, 10, 0, and -10 degrees C) in freely moving conditions. In response to Ta of 0 degrees C for the mice trapped in winter when their mean local habitat temperature (Te) were lower than 0 degrees C, there was a significant inverse correlation between VO2 and Te (r = -0.70, p less than 0.001) whereas no significant correlation (r = 0.23) was observed in the mice trapped in other seasons when Te was higher than 0 degrees C. The correlation between Tco and Te was significant (r = -0.66, p less than 0.001) over the entire range of Te. The pelt weight of the mice trapped at Te higher than 0 degrees C had a significant inverse correlation with Te (r = -0.65, p less than 0.001), but not in the mice trapped at Te lower than 0 degrees C. After measurement of VO2 and Tco at Ta of 0 degrees C, the mice who had lived in colder habitats (below 0 degrees C) showed 0% mortality, whereas the mortality of the populations which had lived in warmer habitats was 13%. These results suggest that, in wood mice, adaptation to severe cold is established by an enhanced thermogenesis and by an increased insulation of the pelt in moderate cold.

Development of a novel polycationic adsorbent for cryogel removal.

Cryogel, prevalent in the plasma of rheumatism patients, is a plasma fibronectin (pFN)-extra domain A containing FN (EDA(+)FN)-fibrinogen (Fbg) complex formed by adding heparin (HP) at a low temperature (4 degrees C). Although EDA(+)FN does not usually exist in normal plasma, its prevalence in rheumatic patients causes cryogelation in plasma. Removal of cryogel is thus a promising and novel approach to treating rheumatism. As HP-EDA(+)FN aggregate, which is induced by the main component of cryogel, is considered to be an anion, cationic materials capable of eliminating this anionic conjugate were innovated in this study. We found that an amino group density of 100-130 micromol/g (dry weight) of adsorbents prompted selective adsorption of the EDA(+)FN-HP complex. Elimination of EDA(+)FN as high as 80% accompanied by removal of the components of total FN (pFN) (10%) and Fbg (10%) in the model patient plasma was established.

Catheterized urine cytology of mucinous carcinoma arising in the renal pelvis. A case report.

BACKGROUND: Primary mucinous carcinoma of the renal pelvis is a rare tumor; therefore, criteria for cytologic diagnosis of this tumor have not been established. CASE: An 81-year-old woman suffered from macrohematuria for six months and was found to have a tumor in the right kidney by radiographic examination. Catheterized urine obtained from the right renal ureter was viscous and contained spherical clusters of cells with occasionally vacuolated, lacy and basophilic cytoplasm. In the small to medium-sized nuclei, chromatin was coarse and granular, and the nuclear membrane was thin and nearly smooth. Large nucleoli were evident in some of the nuclei. These findings were consistent with adenocarcinoma possibly of mucinous type. CONCLUSION: Preoperative diagnosis of mucinous carcinoma is possible by cytologic findings of catheterized urine together with clinical data.

Poroid hidradenoma. Report of a case with cytologic findings on fine needle aspiration.

BACKGROUND: Fine needle aspiration cytology (FNAC) can be used for establishing a diagnosis of cutaneous lesions, especially in cases with cyst formation. Poroid hidradenoma is eccrine neoplasm with both solid and cystic components. CASE: A 77-year-old female presented with a slightly elevated nodule in the skin on her left elbow. The tumor was well demarcated, 2.7 x 2.4 cm and soft, and overlying skin was slightly reddish. FNAC revealed two types of cell: one had abundant cytoplasm in which small to large, occasionally multinucleated nuclei with small but distinct nucleoli were evident. Chromatin was finely granular, and nuclear membrane was thin and almost smooth. Another type of cell had scanty cytoplasm and a round to oval nucleus with small but prominent nucleoli. Histologic diagnosis was poroid hidradenoma. CONCLUSION: FNAC can be useful for diagnosing intradermal cystic lesions before surgical resection.

Reduction of EDA(+) fibronectin and its clinical importance on cryofiltration.

Cryofiltration (CRYO) removes cryogel, which is a combination of fibrinogen (Fbg) and fibronectin (FN), containing pathological substances. The purpose of this study was to measure cryogel EDA(+) FN and study the relationship between EDA(+) FN and clinical symptoms in patients with rheumatoid arthritis, SLE and polymyositis. Cryogel contains 51 times more EDA(+) FN than plasma. The patients with rheumatoid arthritis showed a high level of EDA(+) FN in their plasma, and the EDA(+) FN level in plasma corresponded with changes in joint pain. We calculated the clearance level at several points in cryofiltration, and the reduction enabled us to evaluate the CRYO device. The EDA(+) FN clearance was 23.3 +/- 6.4 ml/min, the pFN clearance 16.5 +/- 4.1 ml/min, and the Fbg clearance 22.9 +/- 5.7 ml/min. As the plasma flow in cryofiltration was 30 ml/min, a clearance of EDA(+) FN and Fbg, approximately 23 ml/min, was obviously high. The study of the plasma level change of EDA(+) FN during cryofiltration revealed a temporary elevation. These results suggest that the EDA(+) FN was most efficiently reduced by cryofiltration, would become a good indicator on plasmapheresis, and might move from other tissues into the blood during cryofiltration.

Artificial conditioner for stored organs.

We have developed an artificial organ conditioning system in order not only to condition but also evaluate the viability for transplant graft of kidneys which have been stored for a long time and damaged by warm ischaemia following cardiac arrest. The conditioning system consisted of an artificial lung, a roller pump, an organ chamber and perfusate. The perfusate was prepared with electrolytes, fluorocarbon, amino acid, glucose, an oxygen scavenger and so on. Conditioning was performed by continuous perfusion under mild hypothermia at 24 degrees C. Mildly damaged kidneys (0 and 30 minutes warm ischaemia rabbit kidneys) were well conditioned but severely damaged kidneys failed to produce urination. Our device successfully exposed the viability of stored kidneys and the successful conditioning of damaged kidneys due to warm ischaemia avoiding transplantation. By establishing our method, the harvesting of kidneys following cardiac arrest will be feasible.

[Statistical studies on the resected gastric cancer of 1538 cases in Hokkaido (author's transl)].

1538 cases with resected gastric cancer in 1974 were registered by the group of surgeons from 76 main hospitals in Hokkaido, who wished to clarify the present situation of the operation of gastric cancer in Hokkaido. The results were as follows; The ratio of male to female was 1.9 to 1.0. The frequency of the resection was 76.9%. The frequency of the early cancer was 13.5%. The frequency of the total gastrectomy was 15.3%. No remarkable differences were found between the above-mentioned results and the results from the whole country. However, the differences between the operable cases estimated by the mortality returns and so forth and the actual cases of the operation amounted to more than 500. Therefore it was clarified that a large number of cases with the gastric cancer for which the operation was indicated failed to be operated. Thus these studies have given out an important subject for the operation on the gastric cancer.

[The significance of blood coagulation test in fulminant hepatic failure].

We treated 12 cases of fulminant hepatic failure with plasma exchange, cryofiltration and hemodialysis. Five cases were survived and 7 cases died. The liver volume measured by volus CT on admission was about 1106 ml in survived cases, and was about 457 ml in death cases. In cases which liver volume was under 600 ml, these prognoses were miserable. AT-III, PT and HPT activities were well correlated with liver volume (r = 0.85) and these activities were good indicator to treat the fulminant hepatic failure. The measurement of FBG and APTT were also useful for the estimation of the prognosis. But they were not becoming of indicator to treatment. We have no expectation to prognosis of the cases which PT, HPT and AT-III were under 15%, 10%, and 25%, respectively.

[A case of essential mesocaval shunt].

Essential mesocaval shunt is very rare entity. A case of chronic cholecystitis with mesocaval shunt from the branch of superior mesenteric vein was reported. This rare hepatofugal pathway was clearly demonstrated by percutaneous transhepatic portography. Selective middle colic venography showed that the branch of this vein was directly connected to the inferior vena cava. After ligation and resection of the shunt, the level of patient's blood ammonia was decreased and the pressure of portal vein was increased. This rare case indicate that the possibility of early diagnosis and interventional radiology for +hepatic encephalopathy.

[Basic and clinical studies on patients who have undergone autologous or allogeneic bone marrow transplantation].

Bone marrow transplantation (BMT) is now increasingly applied not only to hematologic disorders such as leukemias and aplastic anemia but also to some of solid tumors. We experienced ten cases of bone marrow harvest and six cases of autologous or allogeneic BMT during the period from June, 1986 to August, 1987. For autologous BMT cases, bone marrow cells were harvested and cryopreserved at -196 degrees C in liquid nitrogen tank and colony forming assays were examined to ascertain the colony forming ability of harvested bone marrow stem cells. Recovery of colony forming ability after cryopreservation in colony forming unit in culture and burst forming unit in erythrocyte is about 63 to 73% and well preserved. Among 6 cases undergone BMT, five cases are patients with advanced solid tumors including breast cancer, seminoma, lung cancer, malignant lymphoma and brain tumor and these 5 cases have undergone autologous BMT. Conditioning regimen for each cancer in autologous BMT differs based upon conventional chemotherapy regimen and two to four fold dose of conventionally used chemotherapy regimen were administered and after two or three days of high dose chemotherapy, cryopreserved bone marrow cells were thawed and infused. Peripheral white blood cells and platelets were recovered to more than 1000/microliters and 5 x 10(4)/microliters, respectively, within 21 days after bone marrow cell infusion. Disappearance or decrease in tumor size was observed in all cases except brain tumor even though those cases were advanced ones. After autologous BMT, recurrence occurred within three to five months in three cases.(ABSTRACT TRUNCATED AT 250 WORDS)