Type IIa early gastric cancer with proliferation of xanthoma cells.

We report a type IIa early gastric cancer associated with xanthoma cell proliferation in a 61-year-old man. The patient was admitted to our hospital because of a gastric polyp detected at a medical checkup. An irregular protruding lesion with xanthoma cell proliferation was detected endoscopically. Histological examination showed a well differentiated tubular adenocarcinoma in the mucosa associated with xanthoma cell proliferation. The distribution of the xanthoma cells in the stroma corresponded closely with that of the cancer cells. Neither atypism nor mitotic figures were recognized in the xanthoma cells. In an immunohistochemical study, almost all the xanthoma cells were stained positive for alpha 1-antitrypsin, while relatively few exhibited positive S-100 protein staining. Specific monocyte chemotactic and activating factor immunoreactivity was present only in the xanthoma cells, and not in the cancer cells. On the basis of these findings, it was speculated that the gastric cancer cells may have caused the xanthoma cell proliferation via an autocrine mechanism i.e., by a chemical mediator acting in a paracrine or juxtacrine manner.

[Establishment and characterization of a human cholangiocellular carcinoma cell line].

A cell line, HuH-28 was established in vitro from a patient with cholangiocellular carcinoma (CCC). This cell line has been in continuous culture over 10 month period with slow growth potential. HuH-28 was composed of spindle-shaped cells as major population besides a small percentage of polygonal-shaped cells. Chromosome number of the cells were distributed near the hypotriploid region on the 3rd passage. HuH-28 cells were not transplantable into nude mice, but secreted some tumor markers including alkaline phosphatase (ALP), gamma glutamyltranspeptidase (GGT), beta 2-microglobulin (BMG), ferritin, elastase-1 and tissue polypeptide antigen (TPA). This HuH-28 cell line will represent a good model for the investigation of carcinogenesis, histogenesis+ and diagnosis of CCC.

[Culture methods of human liver cells].

Attempts to isolate and cultivate human liver cells have been described. Many viable liver cells have been obtained when dissociated with collagenase followed by dispase. The morphology and differentiated functions have been maintained for more than 3 weeks when human fetal liver cells were cultured not only in medium containing 10(-3) M hydrocortisone, but also on collagen gel substrates with 5 x 10(-7) M hydrocortisone. The colony-forming capacity of primary cultured fetal human livers has also been described in the presence of conditioned medium.

[Studies on AFP-producing capacity and some other properties of human hepatoma cells treated with various anticancer drugs].

The effect of various anticancer drugs on alpha-fetoprotein (AFP) secretion and some other properties of human hepatoma cells was investigated in vitro with the following results. (1) There was a high correlation between AFP secretion and cell number after treatment of human hepatoma cells with anticancer drugs and the amounts of AFP secreted per 10(4) cells per 72 hours (AFP-secreting capacity) were not affected within therapeutically achievable concentrations (TAC). (2) The AFP-secreting capacity was affected with some exceptions in the cells treated with higher concentration of drugs than TAC. Furthermore, chromosomal and morphological aberrations in the similarly treated cells, were also observed, suggesting the relationship between the change of AFP-producing capacity and that of some other properties.

[Tissue culture course of a human hepatoma cell line].

A cell line, HuH-33 was cultured in vitro from a patient with hepatocellular carcinoma. This cell line has been in continuous culture over 12 month period with slow growth potential. HuH-33 was composed spindle-or polygonal-shaped cells as a major population. Chromosome number of the cells were widely distributed even in the primary culture. HuH-33 was transplantable into nude mice and secreted alpha-fetoprotein, albumin, beta 2 microglobulin, ferritin and tissue polypeptide antigen.

Partial adenine phosphoribosyltransferase deficiency detected by ureterolithiasis.

A 30-year-old woman was admitted to our hospital because of recurrent ureterolithiasis. She was suspected of having adenine phosphoribosyltransferase (APRT) deficiency based on the presence of 2,8-dihydroxyadenine (DHA) crystals in her urinary sediment, infrared spectrophotometric analysis of the excreted stone, and then the definitive diagnosis by gene analysis. A pedigree study indicated only a slight possibility of this disease in the family. From these results, we consider that urinary sediment and stone analysis should be used for screening while gene analysis should be employed for definitive diagnosis of APRT deficiency, so that the complications of this condition can be prevented.

Examination of HeLa cell contamination of human cell lines derived from primary hepatomas using glucose-6-phosphate dehydrogenase and lactate dehydrogenase isozymes.

Isozyme patterns of glucose-6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) in human cell lines derived from primary hepatomas were compared with those in HeLa cells. Some cell lines derived from primary hepatomas having type B G6PD showed one or two isozymes of LDH. On the other hand, HeLa cells having type A G6PD showed four LDH isozymes. These findings suggest that not only G6PD, but also LDH may be useful for the detection of HeLa cell contamination of a culture in some cases.

Effect of various factors and substrates on the growth of a human hepatoblastoma cell line, HuH-6 in a serum-free medium.

The effect of various factors and substrates on the growth of a human hepatoblastoma cell line, HuH-6, which was inoculated at low density in a serum-free medium was examined. Several supplements were required to enhance cell growth of HuH-6. These included cholera toxin (CT), glucagon (Glu) and selenium (Se). Type IV collagen (C-IV) provided the most conductive environment tested for cell growth. These results suggest that CT, Glu, Se, and C-IV are important stimulators for the continuous growth of HuH-6 in a serum-free medium at low density.

delta-Aminolevulinic acid dehydratase deficiency porphyria (ADP) with syndrome of inappropriate secretion of antidiuretic hormone (SIADH) in a 69-year-old woman.

delta-Aminolevulinic acid dehydratase deficiency porphyria (ALAD porphyria, ADP) with syndrome of inappropriate secretion of antidiuretic hormone (SIADH) in a 69-year-old woman is reported. The patient was admitted to our hospital complaining of slight cough with low-grade fever, and treated with piperacillin sodium, resulting in complete resolution of the symptoms, following a diagnosis of bronchopneumonia. Thereafter, however, she began to complain of vomiting, abdominal pain, facial numbness and paresis of the extremities with gait disturbance, and became comatose with hyponatremia (serum Na concentration 119 mEq/L) in a few days. Laboratory tests revealed an antidiuretic hormone (ADH) level of 13.5 pg/mL, plasma osmolality 218 mOsm/KgH2O, urinary osmolality 429 mOsm/KgH20, urinary Na concentration > 20 mEq/L, and no abnormalities of thyroid, adrenal or renal function. Neither edema nor dehydration was evident. These data indicated the presence of SIADH. No abnormalities suggestive of malignant or infectious diseases such as lung cancer, pneumonia and Guillain-Barré syndrome were evident from laboratory and roentgenographic findings. As the cause of SIADH, therefore, porphyria was suspected. Metabolites and activities of enzymes in the heme biosynthetic pathway were examined, and very low activity of delta-aminolevulinic acid dehydratase (ALA-D) (0.14 mumol PBG/mL RBC/h) was found. The patient was neither an alcoholic nor a heavy smoker, and she had no past history of heavy metal intoxication, photosensitivity or tyrosinemia. On the basis of these data and clinical features, she was diagnosed as having ADP. We consider this to be the first case of ADP reported in Japan.

Effect of fish oil and safflower oil in common Japanese diet on human plasma fatty acid composition.

The influence of fish oil and safflower oil contained in the common Japanese diet as the main dietary polyunsaturated fatty acid source on plasma fatty acids in ten female student volunteers (21-22 years old) was investigated. The subjects were divided into two groups and fed the experimental diets for five days. The total daily fat intake in the fish diet and safflower oil diet was 54.4 g and 56.2 g, respectively, and the fat derived from fish and safflower oil was 16 g and 23 g, respectively. The proportion of linoleic acid was reduced in the plasma of subjects fed the fish diet and increased in the plasma of subjects fed the safflower oil diet. The plasma levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were significantly elevated in the fish diet group. The ratio of EPA/arachidonic acid (AA) was higher, and those of n-6/n-3 and n-9/n-3 were lower in the plasma of subjects fed the fish diet when compared to the results obtained from plasma of subjects fed the safflower oil diet. From these results, it seems likely that fish oil in the common Japanese diet is a favorable source of plasma EPA and DHA even in such short term supplementation and with such a small amount of daily consumption.

[Laboratory and clinical studies of flomoxef].

Laboratory and clinical studies were performed on a new oxacephem antibiotic, flomoxef (FMOX, 6315-S). In vitro antibacterial activity of FMOX was evaluated in comparison to latamoxef (LMOX), cefmetazole (CMZ), cefazolin (CEZ) using clinically isolated strains of Gram-negative and Gram-positive bacteria. Antibacterial activities of FMOX were stronger than LMOX, CMZ, CEZ against Escherichia coli, Klebsiella but only slightly effective against Staphylococcus aureus. This antibiotic drug was administered to 5 patients consisting of 2 cases with pneumonia, one each with pyelonephritis, chronic bronchitis and urinary tract infection. The drug was given in 1 g drip infusion twice a day for 8 to 13 days. Clinical efficacies of FMOX were excellent in 1 case, good in 2, fair in 1, and unevaluable in 1. As for bacteriological effect of FMOX, organisms were eradicated in 3 cases. No side effect was noted and there was no abnormal change in laboratory findings.

[Management of cesarean section under replacement therapy with factor VIII concentrates in a pregnant case with congenital combined deficiency of factor V and factor VIII].

The congenital combined deficiency of Factor V and Factor VIII, a rare bleeding disorder, was identified in a 25-year-old woman. She was admitted to our hospital with a complaint of genital bleeding. Her prothrombin time and activated partial thromboplastin time were prolonged. She had low levels of Factor V coagulant activity (F. V:C) 14%, and Factor VIII coagulant activity (F. VIII:C), 12%, and normal levels of von Willebrand factor antigen (vWF:Ag), ristocetin cofactor (Rcof) and Protein C antigen. Her Protein C inhibitor level was slightly low. Her Rcof, vWF:Ag and F. VIII:C were elevated following administration of 1-deamino-8-D-arginine-vasopressin (DDAVP), but her F. V:C remained unchanged. Four years later, her F. VIII:C rose to 70% during the course of her pregnancy, but her F. V:C value remained low. It was expected that the vaginal delivery would be possible at the termination of pregnancy. Premature rupture of the membranes and an anomaly of rotation appeared in the course of delivery, however, and cesarean section was accomplished without excess bleeding under replacement therapy with Factor VIII concentrates. These findings suggested that DDAVP and Factor VIII concentrates were useful for management of her delivery. However the mechanisms of the rise of plasma F. VIII:C during pregnancy in a case with congenital combined deficiency of Factor V and Factor VIII are unclear.

[Prolymphocytic leukemia associated with serum hypo IgA].

A 69-year-old man was admitted for the evaluation of leukocytosis with atypical cells. Physical examination revealed marked hepatosplenomegaly. The peripheral blood demonstrated Hb 10.1 g/dl, platelet 13.6 x 10(4)/microliters, and WBC 14200/microliters with 76% lymphoid cells. Bone marrow showed 52.4% lymphoid cells. These cells had a nucleus which was relatively large with a coarse chromatin structure and one prominent nucleolus. Under electron microscopy, these cells had a narrow cytoplasm containing a few mitochondria with some microvilli. The surface of these cells was positive for Ia, B1, B2, C3R, and had markedly elevated IgM-K and IgD-K surface immunoglobulins. Levels of IgG, IgA and IgM were 1140 mg/dl, 53 mg/dl, and 198 mg/dl respectively. He was diagnosed as having B-PLL, and was treated with vincristine, cytarabine and prednisolone. Since B-CLL frequently in accompanied by reduced levels of one or several immunoglobulins, and the most significant is the decrease of IgA, it is speculated from our case that B-PLL is very similar to B-CLL in the abnormalities of B cell function.

[B-chronic lymphocytic leukemia/prolymphocytic leukemia (CLL/PL)--a case report].

A 48-year-old male was admitted to our hospital on April 20, 1989 because of general fatigue and abdominal fullness. Physical examination showed hepatomegaly, massive splenomegaly, and systemic lymphadenopathy. Hematological findings revealed WBC 73,000/microliters, RBC 289 x 10(4)/microliters, Hb 8.0g/dl, and platelet 9.1 x 10(4)/microliters. WBC differential count demonstrated a mixture of 63% matured small lymphocytes and 32% prolymphocytoid cells. Bone marrow aspiration was unsuccessful with a dry tap. Surface marker analysis of peripheral blood lymphoid cells disclosed that they were positive for anti-HLA-DR, CD 5, CD 19, CD 20, CD 21, CD 25, Sm-IgM, Sm-IgD, and Sm-K. He was diagnosed as B-CLL/PL, and treated with VEPA with partial remission. CLL/PL which was advocated by Melo in 1986 is regarded as a distinct clinical entity intermediate between CLL and PLL in clinical and laboratory features. Our case is interesting with regard to good response to combination chemotherapy, though most cases of CLL/PL have a resistance to standard chemotherapy.

[Emergency coronary artery bypass grafting (CABG) in a case of coronary occlusion secondary to long segmental dissection of left anterior descending artery after percutaneous transluminal coronary angioplasty (PTCA)].

A 60-year-old man with unstable angina underwent PTCA at left anterior descending artery (LAD) #6, which had stenosis of 90%. Subsequently, a long segmental dissection formed from #6 to #7 and the patients was scheduled for CABG. On the day before the expected date of CABG, he had an angina attack and a 12 lead electrocardiogram (ECG) showed ST segment and T wave elevation in V1-3. Coronary angiograms revealed 99% stenosis in LAD #6 and immediately rePTCA was performed at the site. RePTCA improved the stenosis to 50%, but ECG showed an inverted T wave in V3,4 and emergency CABG was performed, with the saphenous vein at LAD #8 and high lateral artery. Some authors have stated that CABG is not useful for long segmental coronary dissection, but we conclude that CABG should be definitely considered in such a case, because this procedure resolves pressure gradients at before-and-after entry and the bypass grafts will be never occluded whenever anastomosis (intima-adventitia-graft) is performed.

Role of the mucus gel layer in the healing of acetic acid-induced gastric ulcers in rats.

Changes in the pH gradient and thickness of the mucus gel layer in the healing of acetic acid-induced gastric ulcers in rats were investigated. The maximum of the pH gradient and the thickness of mucus gel layer at the edge of ulcers and in the regenerated mucosa were higher, decreased gradually with the healing of the ulcers and finally reached normal values. This indicates that mucus and bicarbonate are secreted actively from the mucosa surrounding an ulcer and that they play an important role in protection of the base of ulcers and in acceleration of the healing of the ulcers.

In vitro measurement of the pH gradient and thickness of the duodenal mucus gel layer in rats.

The mucus bicarbonate barrier of the duodenum has lately been reported to be one of the most important defensive factors for the duodenal mucosa. We established an in vitro system for evaluating the mucus bicarbonate barrier in the rat duodenum. Our method allows direct measurement of the pH gradient as well as the thickness of the mucus gel layer of the rat duodenum in vitro. The obtained results suggest that alkali secretion in response to acid-loading as well as the thickness of the duodenal mucus gel layer are greater than those in the stomach.

Effect of the luminal hydrogen ion on alkali and mucus secretion in the rat stomach.

This study was designed to investigate the effect of luminal H+ on alkali and mucus secretion in the isolated rat stomach. At various luminal pHs, the thickness and the pH gradient of the mucus gel layer were measured. The maximum pH and calculated alkali secretion showed that the latter was stimulated in parallel with an increase in luminal H+ concentration, especially when the luminal pH became less than 3.5. Mucus secretion, however, was not significantly affected by the luminal H+ concentration. These results suggest that alkali and mucus secretion in the stomach are not regulated by the same mechanism, but different ones.

[Phosphoenolpyruvate:carbohydrate phosphotransferase systems in Enterococcus faecalis].

A wild-type strain of Enterococcus faecalis and its mutants resistant to 2-deoxy-D-glucose (2DG) were examined for the presence of phosphoenolpyruvate:carbohydrate phosphotransferase systems (PTSs) with 12 carbohydrates, which were utilized by the organism, as the substrates. The wild-type strain possessed a constitutive mannose-PTS, which was reactive with glucose, mannose, glucosamine, 2DG and fructose. This activity was absent in the mutants. No independent glucose- or fructose-PTS was found in the mannose-PTS-defective mutants. The mutants, however, showed a low level of a constitutive PTS activity with maltose, suggesting the existence of an independent maltose-PTS in the organism. Both wild-type and mutant strains possessed inducible lactose-, mannitol-, and trehalose-PTSs. Lactose-PTS was induced by either lactose or galactose in the parent, but only by lactose in the mutants. The lactose-PTS was not reactive with galactose, and no separate galactose-PTS was present. These observations suggest that the inducer for lactose-PTS, probably being galactose 6-phosphate, may not be formed from galactose in the organism when the constitutive mannose-PTS is lost by mutation.