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In patients with hepatocellular carcinoma (HCC), natural killer (NK) cell activity decreases significantly, and the reduced activity may be associated with the progression of HCC. In this study we evaluated the effects of pulsing with interleukin (IL)-2 and/or IL-12 on the activation of freshly isolated peripheral blood lymphocytes (PBL) derived from patients with HCC. PBL obtained from 9 HCC patients, 4 liver cirrhosis patients, and 9 normal subjects were cultured in the presence of IL-2 and/or IL-12. After 24 h of incubation, the levels of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha presented in the supernatants were determined by enzyme-linked immunosorbent assay (ELISA). The IFN-gamma and TNF-alpha production of PBL pulsed by a combination of IL-2 and IL-12 was significantly higher than those of PBL stimulated by either IL-2 or IL-12 alone. The mRNA encoding perforin, granzyme B, as well as IFN-gamma and TNF-alpha, were markedly enhanced in PBL stimulated with a combination of IL-12 and IL-2. The pulsing procedure of IL-12 in combination with IL-2 resulted in the increase of IFN-gamma and TNF-alpha, and the expression of perforin and granzyme B mRNA in PBL obtained from HCC patients, as well as in those obtained from normal subjects. These results indicate that adoptive immunotherapy based on PBL pulsed with a combination of IL-2 and IL-12 may be a promising adjunctive strategy for HCC treatment.

We studied the association of gamma-glutamyltransferase (GGT) and other serum markers of liver injury with daily alcohol consumption in a healthy population of 1,043 Japanese males. A positive correlation between daily alcohol consumption and biochemical markers, such as log GGT (r = 0.432), log AST (r = 0.244) or log LAP (r = 0.246), was seen in all drinkers. However, there was a negative correlation, such as log GGT (r = -0.434), log AST (r = -0.424) or log LAP (r = -0.430), in heavy drinkers who consumed more than 70 g ethanol a day. On the other hand, a positive correlation, such as log GGT (r = 0.426), log AST (r = 0.247) or log LAP (r = 0.216) was found in moderate drinkers who consumed less than 70 g ethanol a day. Interestingly, there was a tendency toward negative association between alcohol consumption and the Tokyo University ALDH2 Phenotype Screening Test (TAST) score in the heavy drinkers, and there was a tendency toward positive association between GGT and TAST score in this group. Our results suggest that there are 2 groups of drinkers, those with elevated GGT (good responders) and those with normal GGT (poor responders) despite heavy drinking.

For the purpose to reveal the relation between cell death and nuclear stainability by supravital staining with basic dyes observations have been made on the cells of bone marrow, peripheral blood and lymph node from anemic and non-anemic rabbit, rat, mouse and chicken, and thymus from young mouse. The cells were stained supravitally in blood serum, isotonic saline, calcium chloride and sucrose solutions with the dyes; brilliant cresyl blue (B. C. B), Nile blue (N. B.), neutral red (N. R.), Janus green (J.G.) and eosin (E.). The following results were obtained: 1. In the presence of blood serum all the living cell nuclei observed were not stained supravitally, except some mature erythroblasts and nucleated red cells. 2. In isotonic saline, CaCl2 and MgCl2 solutions all the erythroid cell nuclei were stained deep by B. C. B., N. B., N. R., slightly by J. G. but not by E. In stainability the younger the cell is the deeper in its nuclear staining. The nuclei of other cell strains were not stained. 3. In isotonic sucrose the nuclei of mature granulocyte were also stained by B. C. B. and N. B. but not by other dyes. The nuclei of lymphoid cells and myeloid cells appeared pale without being stained by any dyes. The nuclei of erythroblasts in sucrose solution were stained deeper with B. C. B. and N. B. than those in isotonic saline. The differences between supravital stainability of the nuclei among the cells belonging to different strain and among those of the same strain but in different maturation stage and the nuclear staining after cell death have been discussed from the possible dissociation of DNA from histone.

In order to ascertain whether black-crowned night herons (BCNH), white heron (Plumed Egrets (PE)) and domestic fowls are infected by JE virus and they serve as infection source ofJE, hemoagglutination inhibiting antibody and its 2·ME sensitive antibody in the sera of these birds were determined. Physico-chemical nature of fowl's antibody of JE produced by natural infection and their maternal antibody in the sera of chicks were examined. The results are briefly summarized as follows. 1) As to the herons captured in Tsudaka Town, two out of six adult night herons and three out of the four chicks showed positive HI reaction. On the other hand, HI reaction in the sera of two adult white herons and three chicks were negative. 2) As to the herons captured in Okayama City, twenty out of thirtytwo adult night herons and seven out of seventy white herons showed positive HI reaction in 1966 around the time when JE was prevalent in Okayama Prefecture. And six out of eleven night herons and one out of seven white herons showing positive HI reaction, responded positively to 2-ME sensitivity test. 3) The results indicate that white herons can be also infection source ofJE though less than in the case of night herons. 4) In the domestic fowls (white leghorn) kept at Takahashi District, eight out of twenty-seven fowls showed positive HI reaction. And six out of seven domestic fowls showing positive HI reaction responded positively to 2-ME sensitive reaction. 5) Transformation of JE antibody in the serum of hen from IgM to IgG was recognized. 6) Domestic chicken's sera having 1 : 640 of HI titer in the original serum and 1 : 320 of HI titer after 2-ME treatment were fractionated by gel filtration on Sephadex G-200 and the antibody activities present in the various fractions were determined. HI antibody activities occurred in both IgM and IgG classes of immunoglobulins. 7) Maternal HI antibodies reacting with JE virus were found in newly hatched domestic chickens from the eggs laid by hens with natural infection ofJE. And half life of HI antibodies in chicks was four days. 8) HI antibodies of JE in the serum of maternal immune-hens and chicken having maternal antibody were located in r-globulin fraction by starch block electrophoresis. 9) The results from 4) to 8) indicate the presence of natural infection ofJE in the domestic fowls. And domestic fowls can be infection source ofJE.

To obtain some information of the biological action of Kankohso 101dinicotinate and Kankohso 301-nicotinate, observations were made on the binding mode of these substances with protein, chondroitin sulfate and nucleic acids and the following results were obtained; 1. Kankohso 10 I-dinicotinate binds reversively with bovine serum albumin or serum r-globulin, resulting in metachromasia. By binding with proteins the absorption maximum of the dye shifts toward the long wave length side and the absorbance decreased distinctly. The data show that there are more than one kind of binding sites and the binding with bovine serum albumin is weak in acidic solution and strong in alkaline solution. 2. Kankohso 10 I-dinicotinate produces strong metachromasia with sodium chondroitin sulfate and the color of the solution changes from violet blue to reddish violet. The absorption maximum at 592 mp. decreases without shifting its wave length ,and the shoulder appears at 555 mp. be. comes distinct peak. The strongest metachromatical changes occurs at the concentration of the chondroitinsulfate whose sulfonate radicals is equal to the molecules of Kankohso 10 I-dinicotinate. 3. Kankohso IOI-dinicotinate produces metachromasia with nucleic acid, where absorption spectrum is shifted toward long wave length and absorbance is decreased at a certain concentration. 4. Kankohso 301.nicotinate binds weakly with bovine serum albumin, the binding of which is reversible and the maximum binding number is 1.1 per molecule of albumin. Metachromasia cannot be produced by binding. Kankohso 30I.nicotinate does not bind with bovine serum γ-globulin. This compund does not produce metachromasia with sodium chondroitin sulfate but produces weak metachromasia with nucleic acid, indicating some affinity to nucleic acid.

Die stark eisenhaltigen Praparate Ferrlecit, Kobaltjerrlecit, Ferrojolsan, Ferrojolcamma und Irradian wurden im Siemens Unterrichtsreaktor SUR 100 des Instituts fur Kernenergetikder Universitat Struttgart und der Technischen Universitat, Berlin mit einem NeutronenfluB von ca. 5.l06n/cm2 sec bestrahlt. Es ollte dabei untersucht werden, wie stark eisenhaltige Praparate mit diesem Reaktor aktiviert werden konnen. Es wird festgestellt, daB Eisen praktisch kaum aktiviert wird. Die Ergebnisse zeigen, daB nur das Kobattjerrlecit und das Irradian starker durch Neutrosen aktiviert werden. Die hohere Aktivitat des Kobaltjerrlecit wird auf die darin enthaltenen Elemente Mangan, Kupfer und Kobalt zuruckgefuhrt, die mit thermischen Neutronen gut aktivierbar sind. Die hohere Aktivitat des Irradian geht hauptsachlich auf β-Zerfalle zuruck. Das Phosphorisotop P-32 tragt einen groβen Anteil dazu bei. De rβ-Untergrund des Irradian bei hohen Energien, der mit einer kleinen Halbwertszeitabklingt, ist durch keines der angefuhrten Elemente im Irradian erklarbar. Der Mn-56Peak im Spektrum ist gut erkennbar, durch den β-Untergrund wird er aber zu einer hoheren Energie hin verschoben.

Les auteurs out effectue après une irradiation totale de 1200r de rats blancs des deux sexes des examens hématologiques à la suite d'irradiations ainsi que des examens physiologiques et des contrôles. Ils n'ont observe de modification importante des facteurs coagulants qu'au troisieme jour; cette modification était maximum avant la mort, c'est-à-dire au stade terminal. Les temps de coagulation naturelle ont beaucoup diminué, de même que ceux de la thrombine et ceux de la thrombine avec le bleu de toluidine, c'est-à-dire que l'héparine libérée ( = antithrombine semblable à l'héparine) a diminue. Pour les facteurs V et VII et en particulier pour la prothrombine on a observe un fort accroissement de la concentration. Les auteurs pensent que ceci est explicable par le fait que la décomposition des tissus pendant l'irradiation entraine la libération de kinase et d'autres activateurs dans la circulation sanguine, ce qui provoque une anoxemie des tissus. D'autres expériences sont en cours en collaboration avec de nombreux spécialistes et instituts.

For the purpose to clarify whether minimal catalatic activity exists in Japanese acatalasemic cells or not and the manner how extrinsic hydrogen peroxide affects the acatalasemic cells, the author performed tissue cultures using the skin specimens from four acatalasemic persons affected with Takahara's disease and studied the nature of these cultured cells. The results are summarized as follows. 1. Between normal and acatalasemic cultured cells, no morphological differences could be seen and the growth rate of these cell-lines was similar to one another. 2. On the activity of succinoxidase and cytochrome oxidase there could be observed no difference between normal and acatalasemic cells. 3. In each acatalasemic cell line the minimal catalatic activity was observed and it seemed that this activity has an important role in decomposing hydrogen peroxide under normal metabolic pathway. 4. After treating with 10-4M hydrogen peroxide, respiratory enzyme activities and the growth rate in the acatalasemic cells were markedly disturbed, while in normal cells these remained almost intact. 5. There could be observed no differences between normal and acatalasemic cultured cells after X-ray irradiation (200 to 600 r) on the succinoxidase activity, catalatic activity and growth rate.

Ultrastructure of microfilaria Brugia malayi was investigated with electron microscope. Microfilariae are covered by a sheath membrane with dense materials on its outer surfaces. The cuticle consists of 3 layers; namely, external cortical, internal cortical and fibrous layer. Beneath these cuticular layers, thin hypodermis is present and the muscle cells are arranged of 4 groups in a crosssection except for the head and tail. A pair of cephalic channel containing several cilial rods opens at the anterior end of the worm. A hook is situated on the anterior edge of one channel orifice, and several spines grow on the opposite side to the hook. Caudal channels paired laterally opening into the both sides of the posterior region differ from cephalic channels by the presence of a single cilial rod. A central canal runs from the buccal cavity to the inner body, and opens into the inner body cell through the filamentous apparatus. The inner body appears to consist of several cells having storage substances and a flat nucleus located on the periphery of the cell. An excretory apparatus, i. e., a cell, is composed of a nucleus and a large vesicle which has many microprojections on the luminal surfaces. The GI cell which occupies the whole width in a cross-section is larger than the R cell. R2-R4 cells appear to be in a close contact with the anal apparatus having many microprojections on the luminal surfaces. These microprojections differ from those of the excretory vesicle in their thickness and length. The characteristic patterns of these organs are compared with other microfilariae.

The mode of lymph flow from the breast to the regional lymph node was studied using 1llSAu colloid, to ascertain the concept of lymph node of Groups 1, 2, and 3. Lymph flow through the internal mammary nodes was found to be abundant. Especially so when 1llSAu colloid was injected into the lower half of the breast, there was observed more abundant flow than that to axillary lymph nodes. Therefore, it was concluded that regardless of the site of occupation of a tumor, internal mammary nodes should be con· sidered to be of Group 1. Lymph flow through the subclavian nodes proved to be always less than that to the axillary nodes or internal mammary nodes. The rate of the lymph flow appearance on the scintigram was about 20%. Whether the subclavian nodes should be classified as Group 1 or 2 should be further studied. The rate of supraclavicular node visualization was below 3%. Therefore, the supraclavicular nodes should be considered to be Group 2. When these results are applied to the current rule of handling of breast cancer, discrepancy occurs in the Stage number and R-numer.

To evaluate urinary albumin index (UAI), the relationship between albumin excretion rate (AER) in the urine stored for 24 h and UAI in the urine collected arbitrarily on the morning of the same day was studied in 123 inpatients. The patients were admitted to our hospital from September 1, 1988 to August 31, 1989, consisting of 67 non-insulin dependent diabetics (Group 1), 40 patients with collagen disease (Group 2), and 16 patients with primary renal disease (Group 3). The relationship between log(e) AER and log(e)UAI was plotted on a graph. Pearson's rank correlation coefficients of Groups 1-3, Group 1, Group 2, and Group 3 were as follows: r = 0.725, r = 0.691, r = 0.855, and r = 0.611, respectively. The formula obtained by using Pearson's rank correlation coefficients to estimate log(e)AER from log(e)UAI in 123 cases of Groups 1-3, 67 cases of Group 1, 40 cases of Group 2, and 16 cases of Group 3 were: log(e)AER/log(e)UAI = 0.815, log(e)AER/log(e)UAI = 0.860, log(e)AER/log(e)UAI = 0.830, log(e)AER/log(e) = 0.722, respectively. In the present study, log(e)UAI was found to correlate well with log(e)AER. As AER is generally accepted to be the most reliable index to know the stage of albuminuria, UAI is considered to be clinically useful.

In Anbetracht der oft schwierigen Umgrenzung des Ekg Bildes erschien es von Interesse, an einem gröβeren Personenkreis mit einem neuen Beobachtungssystem Untersuchungen unter dem Gesichtspunkt anzustellen, ob Herzgesunde charakteristische Veränderungen der Herzstromkurve aufweisen. In der vorliegenden Arbeit wird im Hinblick auf diese Fragestellung über Rhythmusstörungen im Tagesablauf berichtet, die bei 100 Herzgesunden jüngeren Erwachsenen während einer kontinuierlichen Magnetbandregistrierung von nahezu 24 Stunden Dauer gefunden wurden. AUβerdem wird das Auftreten der RhythmusstOrungen bei "Herzgesunden", mit dem 35 Patienten der "alten Gruppe", die anamnestisch und physiologisch an keiner Herzerkrankung leiden und mit 100 "Koronarleiden" verglichen. 1. Die beobachteten Rhythmusstörungen waren bei allen Gruppen .sehr ausgeprägt, d. h. jede fUr sich 55%, 100%, 97%. 2. Die am häufigsten beobachteten Abweichungen bei allen Gruppen waren Extrasystolen (ES) d. h. 49%, 100%, 97 %. 3. Häufiger fanden sich mehr supraventribuläre ES als ventrikuläre bei Herzgenuden, (31: 29 FäIle), bei der "alten Gruppe" (29: 22 FäIle), nicht dagegen bei "Koronarleiden" (67: 92 FäIle). 4. Von allen ES fanden sich die Kammer-ES am häufigsten. (29, 22, 92 FäIle). 5. Der Schrittmacherwechsel trat nur bei einigen der Herzgesunden und zwar ausschlieBlich nachts während des Schlafes auf. Diese Tatsache müβte auch seine funktionelle Genese bestätigen. 6. Vorhoffimmern und paroxysmale Tachykardien traten bei Herzgesunden nie auf. Derartige Erscheinugen können auβerordentlich bedeutungsvoll sein. 7. Sicherlich ist das Vorhandensein eines Av-Blockes 2. bzw. 3. Grades ein Hinweis auf die Notwendigkeit einer sorgfältigen und kritischen Untersuchung. 8. Wenngleich je nach den Umständen und der Tageszeit des Auftretens für die meisten Kurvenabweichungen und RhythmusstOrungen Änderungen des vegetativen Tonus von wesentlicher Bedeutung sein durften, fanden sie sich bei Personen mit erhöhrter vegetativer Labilität dennoch nicht gehäuft. Aus den Ergebnissen wird der Schluβ gezogen, daβ in der Bewertung der hier beobachteten, meist flüchtigen Veränderungen bei der Ekg-Beurteilung Zurückhaltung geboten ist, Kontroll- und Funktions-Ekg sollten häufiger als üblich zur Abklärung herangezogen werden, ehe die Diagnose einer pathologischen Abweichung gestellt wird.

1. In order to obtain direct evidence for the enzymatic identification of the head-pieces of the elementary particles in the inner mitochondrial membrane, the head-pieces were detached by sonication from the isolated inner membrane of beef heart mitochondria, purified by pursuing the particles with the electron microscope, and analyzed for enzymatic properties. 2. Electron microscope examination revealed that the isolated headpieces are the spherical particles about 90À in diameter which are quite similar in appearance to the head-pieces of the elementary particles lining the inner mitochondrial membranes. 3. The head-pieces are identified as ATPase sensitive to oligomysin when attached by stalks to the membrane, and become insensitive when detached or purified from the membrane. 4. The head-piece is labile to cold with respect to ATPase activity and morphology.