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Forty-five patients with acute leukemia were compared on cellular immunity measures versus prognosis. The patients were treated according a multicombination therapy protocol. The purified protein derivative (PPD) test and dinitrochlorobenzene (DNCB) test on admission indicated low positive percentages. In acute non-lymphocytic leukemia (ANLL) patients, the 50% survival durations were 11 months in the PPD positive group and 6 months in the PPD negative group. In acute lymphocytic leukemia (ALL) patients, the 50% survival durations were 21 months in the PPD positive group and 13 months in the PPD negative group. Peripheral lymphocyte blastogenesis by phytohemagglutinin (PHA) stimulation was examined at various clinical stages. The stimulation indices were generally low, and no correlation was found between the PHA test and clinical stages. These cellular immunity measures appeared to reflect one aspect of the clinical condition in acute leukemia patients, and further studies are needed for predicting prognosis.

Electron microscope observations were conducted on the relationship between mitochondria and inclusion body in mice spheroid alveolar epithelial cells after injection of trypan blue, an acidic dye and Alcian blue 8GS, a basic dye, by vital staining procedures. When both dyes were injected, the mitochondria of the spheroid alveolar epithelial cell became degenerated; however, in injection of only trypan blue, the cristae showed an increase in electron density. In injection on only Alcian blue 8GS, the cristae showed negative contrast. In most cases the trypan blue particles did not enter into mitochondria, whereas particles of Alcian blue 8GS sometimes entered into the mitochondria. When trypan blue particles entered mitochondria, deposits were not evident in the inclusion body, whereas when Alcian blue particles entered mitochondria deposits were seen in the inclusion body. In both of these cases only a few inclusion bodies were formed so that only traces or no inclusion bodies with vacuolar appearance were observed. From these findings it is suggested that mitochondria maybe convert to inclusion bodies.

The relationship between alveolar macrophages and spheroid alveolar epithelial cells was studied with the electron microscope after injection of squid-ink solution into the trachea of the mouse. At 20 hours after injection of squid-ink solution slight degeneration was evident in alveolar macrophages with sepia-melanin particles being phagocytized with partial digestion by lysosmes. Furthermore, hardly any changes were seen in mitochondria and inclusion bodies of the spheroid alveolar epithelial cells. In contrast, at one week after injection of squid-ink solution, almost all alveolar macrophages were degenerated with destruction of the ectoplasm in which the ingested sepia-melanin particles were digested by lysosomes into fine particles, and the mitochondria of spheroid alveolar epithelial cells were degenerated and the inclusion bodies were hardly formed. At three weeks after injection of squid-ink solution, alveolar macrophages as well as speroid alveolar epithelial cells showed almost complete recovery of functional structure. As the phagocyte in the alveolar space, neutrophile leucocytes were also observed in addition to the so-called alveolar macrophage.

Tissue localization of a subcomponent of the first component of complement (CLq) was examined in one postmortem case of HBs antigen (HBs Ag) positive hepatocellular carcinoma and in six cases of chronic hepatitis from liver biopsy specimens. The direct immunofluorescent method was used after fixation with 2% para-formaldehyde in concentrated ammonium sulfate. CLq localization was found in collagen fibers and the cytoplasm of fibroblasts in the connective tissues of specimens examined. The localization was particularly marked in the region of the fundal glands of the gastric wall. Apart from collagen fibers, other sites of localization included the surface membrane of lymphocytes, especially those cells of the mesenteric lymph nodes. In HBs Ag positive specimens, immune deposit-like substances appeared localized intra-hepatically and in the renal glomeruli. Since C3 and C4 were identified concomitantly, it indicates that these substances were indeed immune diposits. Despite the finding that C3 and C4 were identified together in the hepatic cell cytoplasm, C1q itself was not demonstrated in all hepatic cell cytoplasms.

Today Vitallium is used for surgical implants. It is a casting alloy which, with advances in casting technology, is also used commercially for making instruments of fairly complex shape. Because of its expense, however, it is not widely used in Japan. Instead, a series of 18-8 Mo alloys are used in Japan even though of insufficient strength. Used over a long period of time in the body, especially for the purpose of preserving structual functions as part of the human skeleton, it often corrodes, resulting in either abnormalities in tissue cells or, because of its insufficient strength, danger of bending and breaking with aging. In spite of a marked advance in fracture treatment, we have hardly any suitable materials for making instruments appropriate to the internal fixation of fractures in Japan. We, therefore, conducted various experiments to develop an alloy with sufficient corrosive resistance and strength that could be formed into a complex shape to take the place of Vitallium alloy, finally succeeding in developing an alloy we call "COP". The characteristic properties of COP may be summarized as follows: 1. The main components are 20% Cr, 20% Ni, 20% Co and 4% Mo aside from 0.2% P. 2. As it contains "P", it shows a marked age-hardening. In its molten state its machinability is excellent, and later it can readily be hardened by heat-treatment. 3. It has not only a marked yield point and tensile strength but also has toughness in elongation and reduction of area, showing a strength which surpasses Vitallium. 4. Its corrosive resistance is great. 5. Its cost is far cheaper than Vitallium.

Further purification and characterization are reported on rat cytosol cornin (RLCC), an antimitotic substance. Fraction I (purified RLCC) was purified more than 10-fold from crude RLCC with Sephadex G-50 column chromatography and showed a remarkable inhibitory effect on division of inseminated sea urchin eggs and mouse fibroblast cells. Fraction I was observed as one spot, and the molecular weight was estimated to be about 25,000 by thin layer gel filtration. Fraction I contained protein (92%) and RNA (8%), but the antimitotic activity was scarcely affected by treatment by pancreatic RNase. The protein of Fraction I was separated into two bands by SDS-polyacrylamide gel electrophoresis, and the molecular weight was estimated as 10,000 and 15,000, respectively. The 50% inhibition dose of Fraction I on the first division of inseminated sea urchin eggs and on proliferation of mouse L cells was about 2.5 X 10(-5) g/ml and 5 X 10(-4) g/ml, respectively. The yield of fraction I was about 35 mg from 100 g rat liver.

The present study was conducted to investigate the usefulness of the direct leucocyte migration agarose method for studying cell-mediated immunity in vitro. Comparative studies of the purified protein derivative (PPD) skin test and the leucocyte migration inhibition test (LMIT) in which PPD was used as test antigen indicated a significant qualitative and a weak quantitative correlation between these two tests. Furthermore a positive correlation was found between the LMIT and the macrophage migration inhibition test (MIT) using ultrasonicated authochthonous tumor antigen. Comparative studies of the LMIT, MIT, PPD skin and DNCB tests on the same patients showed that cases responding positively to the the PPD skin and DNCB tests tended to respond positively to the LMIT and MIT. Patients with digestive organ cancers were examined by the LMIT. With the advance of cancer, decreased positive test test rates were found. After gastric cancer operations the LMIT findings were divided into two groups: one type changed from positive to negative, and the other type changed from negative to positive. The former response was suggestive of a successful operation, and the latter response was suggestive of a non-curative operation. These results indicated that the direct leucocyte migration inhibition agarose test was useful investigating cell-mediated immunity.

Twenty cases of suspected drug induced liver injury (16 cases of which satisfied the criteria for at least a query positive diagnosis as based on the Fourth Congress of "Drugs and the Liver" in Japan) were studied by the whole blood culture technique of lumphocyte blast transformation. The positive rate with this technique was 10%, and no more than 15% even with the addition of one query positive. One reason for the low positive rate was that there was not only an allergic mechanism at work in the study group but that liver injury due to direct cytotoxicity of the drug was involved also. For a drug such as chlorpromazine with strong cytotoxicity for lymphocytes, it was difficult to demonstrate a relationship between allergic mechanisms and the drug with this method.

1. Three cases of acute rejection were detected by macrophage migration inhibition tests (MIT) conducted directly on seven patients who had received renal allografts. The macrophage migration inhibitory factor (MIF) activity was positive in all cases 1-2 days before the appearance of acute rejection. 2. After the administration of a high dose of Solu-Medrol (1g/day for 3 days) to suppress the acute rejection, MIF activity recovered to its normal level 3 days later. These findings seem to indicate that MIT yields immunologically useful criteria for the early detection of an acute rejection.

Mice were trained in an avoidance learning task. The incorporation of 3H-leucine into the hippocampal regions of trained mice was higher than that of control mice. When mice were injected with cycloheximide, a strong inhibitor of protein synthesis, impairment was evident in acquisition of learning. Cycloheximide produced morphological changes in mitochondria and microtubules of some brain axons. It is suggested that the cycloheximide-induced learning impairment may be due to the blocking of the synthesis of the specific protein necessary of neutral conductivity.

The relationship between immune insulitis and glucose tolerance was investigated in three groups of mice following active immunization with different components of bovine pancreatic hormone. An abnormal blood glucose level was observed in the three groups ranging from 33.3% to 87.5% of sensitized mice. A relationship was not present between the glucose tolerance response and the presence of insulitis or anti-insulin antibody in the blood of sensitized mice. However, all sensitized mice with a marked decrease in glucose tolerance were found to have insulitis. In animals without established insulitis and with no demonstrable anti-insulin antibody, abnormal glucose tolerance was noted. This latter condition occurred more frequently with recrystallized insulin than with a-component and did not occur with monocomponent insulin. These findings seemed to indicate that two distinct processes involving some circulating antibodies with anti-insulin antibody and insulitis might be involved in the development of the observed glucose tolerance abnormality.

Cell-mediated immunity was studied in 23 cases of advanced gastrointestinal cancer. The patients received levamisole at 150 mg/day for three consecutive days each week for four weeks. In cases at the terminal stage of gastrointestinal cancer, the blastformation rate of peripheral blood lymphocytes against phytohemagglutinin (PHA) after the administration of levamisole showed a slight increase, but cases with blastformation rates over 40% increased markedly three or four weeks after the initial administration of levamisole. The peripheral blood lymphocyte count showed little change in these cases.

Plasma postheparin lipolytic activity (PHLA) was measured on 50 patients with systemic lupus erythematosus (SLE). Plasma PHLA was significantly decreased in SLE patients. This decrease was most striking in the acute phase of the disease. There was a close relationship between decreased PHLA and immunologic factors indicative of the acute phase of SLE. These immunologic factors included shaggy antinuclear antibody pattern, low serum complement titer, high DNA antibody titer, mixed cryoglobulin and lumpy glomerular pattern by immunofluorescent staining.

The microfilaria of Brugia pahangi obtained from an experimentally infected dog were observed with the electron microscope. The sheath was composed of small granules and was covered with electron-dense particles on the outer surface and with small granules on the inner surface. The cuticle was composed of an outermost layer, a trilaminate membrane and an inner layer. The hypodermis was composed of four components as in the adult stage (two small ones on the lateral sides, two large ones on the dorsal and ventral sides). The muscle cells comprised a single layer under the hypodermis on the dorsal and ventral sides. On each side, two muscle cells usually appeared in a transverse section. The thick myofilament was surrounded with 8 to 12 thin myofilaments. Dense bodies were present around the cephalic space. In the cells of the nuclei column, the cytoplasm was very narrow, and the electron-dense nucleus close to each other. The cuticular central canal was connected to the buccal cavity and to the inner body. A sponge-like structure was seen at the junctional part of the canal and the inner body. The inner body showed a homogeneous granular appearance. Eight cephalic papillae were observed at the head tip. Two amphids, each having more than eight cilium-like structures, were connected with the nerve elements and open in the head part. Two phasmids, each having one ciliumlike structure, opened in the caudal part. Two types of neurosecretory granules were observed in the nerve ring and the dorsal and ventral longitudinal nerves were clear except in the anterior and the posterior part of the worm. The excretory and the anal vesicles had contacts with thin and thick cytoplasmic processes respectively, and these vesicles opened to the exterior. The nuclei of the G cell and R cells showed similar electron-density. Lamellate structures were present in the muscle and the hypodermis.

The precipitation reaction of bovine serum albumin coupled with p-azophenylleucine with homologous antibody was inhibited by several structurally related haptens. The isobutyl group substituent on alpha-carbon atom of the leucine residue contributed more than -5.8 Kcal/mol to the free energy of binding. This value was consistent with the free energy change expected from the transfer of n-butane from an aqueous environment to liquid n-butane. The observed contribution was explained, in terms of the hydrophobic interaction of the isobutyl group with the antigen binding site of the antibody molecule. These results were also compared with other hapten-antibody systems.

With the electron microscopic and the negative staining method, the glomerular basement membrane of human and bovine kidneys was shown to have a definite fine meshwork structure. The pores of the meshwork of bovine glomerular basement membrane appeared to be pentagonal or hexagonal in shape. Strands of the meshwork branched three-dimensionally and made up the whole basement membrane. The portion of the sgrand between two neighboring branches was presumed to be a structural unit of the basement membrane. Glomerular basement membrane in man showed a structure similar to that seen in cattle, although the pore size of the meshwork was smaller in man than in cattle.

Gastrointestinal cancer patients were followed up for up to 30 months postoperatively and their clinical status related to a parameter of nonspecific immunity, the blastformation rate of peripheral blood lymphocytes against phytohemagglutinin. By the fourth postoperative week, the blastformation rate had recovered from the effect of the operation. In patients who had undergone curative resection, the postoperative level rose to exceed the preoperative level, whereas whereas in those in whom resection had not been possible, the blastformation rate failed to show this rise by the fourth week, and continued at the decreased immediate postoperative level. Results for long-term follow-up (30 months postoperatively) showed that the blastformation rate continued at high levels (almost all over 40%) in cases of curative resection without recurrence, but remained low (under 40%) in those in which the tumor could not be removed. The 40% level of the blastformation rate test thus correlated well with the prognosis. The blastformation rate, therefore, proved a very good parameter for following the pre-and post-operative clinical course of gastrointestinal cancer patients.

In vitro and in vivo testosterone metabolism was studied using rat skin. The in vitro stoichiometric studies strongly suggested the existence in the skin of 5a-reductase and other enzymes involved in testosterone metabolism. The predominant metabolites were 5adihydrotestosterone, androstandiol and 6.4-androstenedione. The rate of testosterone metabolism in vitro was higher in preputial skin than in dorsal skin. The main metabolites were androstandiol and 6 4-androstenedione in vivo whereas dihydrotestosterone and androstandiol were predominant in vitro. .6.4-Pathway is considered to be more active in vivo in adults than in newborns. The difference in the in vivo uptake increase of testosterone-3H after castration suggests the possible existence in the organs and skin areas of androgen pools of different sizes. The in vivo uptake and metabolism of testosterone were examined in the nuclei of newborn rats. Dihydrotestosterone and androstandiol detected in the nuclei of preputial skin, dorsal skin and liver were 42.8%, 24.3% and 27.8%, of total radioactivity incorporated into the respective organs. The liver incorporated only a small amount of radioactivity compared with the skin.

The surface of Gross virus-induced murine lymphoblasts and C-type virus particles budding from these cells were investigated under the scanning electron microscope (SEM). The cells appeared spindle-shaped or roughly-rounded with extensive surface features consisting of microvilli, blebs and ruffled membranes. C-type virus particles were detected on the cell membrane as small spherical particles, distinguishable from the microvilli. Clustered virions were observed in some cases. However, the distribution of virions appeared to be random. The surface of the virion was smooth and had no globular units at high magnification. These morphological observations were confirmed in ultrathin sections.