Search

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.

Effect of inorganic phosphate on ferrous ion- and ascorbate-induced lipid. peroxidations of isolated rat liver mitochondria was investigated. As a result it has been shown that phosphate accelerates the ferrous ion.induced lipid peroxidation; namely, phos. phate shortens the induction lag period of the lipid peroxidation reaction but the malondialdehyde after onset of its production is yielded at the same rate in various concentrations of phosphate. On the other hand, phosphate inhibits ascorbate.induced lipid peroxidation. There are stoichiometric interactions between the concentration of phos. phate and the induction period. Oxygen uptake by mitochondria was observed in the presence of both ferrous ion and phosphate at initial step of the reaction without being accompanied by malondialdehyde production, and afterwards there occurred malondialdehyde production with rapid rate of the oxygen uptake. Possible mechanisms and interactions among ferrous ion, ascorbate and phosphate were discussed.

For predicting median date or incidence rate in the prevalence of japanese encephalitis, the authors considered the 34 factors; namely, the climate, latitude, longitude, and date showing immunological positivity of hemoagglutination inhibiting reaction on 50% the number of swine, etc. To make the mean square residual the smallest which yields, in the case of calculating with the multiple regression equation, the most important and meritorious factors were selected from the factors mentioned above by the voluntary selection rule devised by us. Multiple regression equations were formulated for them. To predict the median date in the prevalence of japanese encephalitis in whole japan, we found that the three factors, i. e. the amount of rainfall in April, the average temperature in March and HI positive rate of swine till the end of july, were essential. And for the foreseeing of incidence rate in Okayama Prefecture, factors concerning mosquitoes were added; this resulted in the useful two factors, namely, common logarithm of the total number of Culex tritaeniorhynchus till july 20th and the rainfall in June.

By dividing at random 14 normal persons into 7 pairs of two individuals each, lymphocytes were isolated from their peripheral blood and taking one of the pairs as stimulating cells or antigens and the others as responding cells, mixed lymphocyte culture (MLC) was carried out. As for the method of MLC we used our MLC method of unidirectional mixed culture with a small amount of lymphocytes in additition of 1% (v /v) PHA.M and cultured for three days, and a widely used conventional method in which 3H.thymidine uptake was the parameter of the blastformation rate and cultured for seven days. In comparing the results of these two groups of MLC the data in six experiments out of the seven coincided. Namely, with 5x 104 cells each of stimulating cell group and responding cell group, it is possible to achieve satisfactory MLC, the culture can be done only for three days without requiring any special technique and the results can be readily evaluated. Therefore, MLC by our simple method would yield satisfactory results in clinics.

The percentages and absolute numbers of gamma delta T cells per CD3 positive cells (T cells) in four different compartments, namely peripheral blood, synovial fluid, synovial membrane and lungs from patients with rheumatoid arthritis (RA) and in peripheral blood from healthy controls were studied by two color flow-cytometric analysis. The percentages (mean +/- SEM = 6.3 +/- 0.8%, n = 22) and absolute numbers (70 +/- 11/microliters, n = 22) of gamma delta T cells in peripheral blood from RA patients were not different from those of 22 age-matched healthy controls (7.5 +/- 0.9%, 81 +/- 17/microliters, respectively). The gamma delta T cells in peripheral blood from 50 RA patients were, however, significantly decreased in negative correlation with the value of CRP as a marker for inflammation, although they had no correlation with the titer of rheumatoid factor as an autoantibody. The percentages of gamma delta T cells in synovial fluid from 10 patients (3.3 +/- 0.5%, n = 10) or in synovial membrane from 5 patients (4.2 +/- 1.9%, n = 5) and in bronchoalveolar lavage fluid from 6 patients (3.6 +/- 0.8%, n = 6) were not different from those in peripheral blood from the same patients. Thus, gamma delta T cells are not the dominant infiltrating T cell subset in the inflammatory sites of RA patients.

Cytochemical observation of the activities of diphosphopyridine nucleotide diaphorase (DPNH-D), triphosphopyridine nucleotide diaphorase (TPNH-D), succinic dehydrogenase (SDH) and a-glycerophosphate dehydrogenase (α-GDH) of Ehrlich ascites carcinoma cells were made and following results were obtained. The smeared cells showed moderate reactions and no marked difference in the intensity among the individual cells. The free floating cells were stained relatively faint but showed the differences in the staining intensity in individual cells. In the presence of benzalkonium, the reaction intensity proved to be intermediate between the smeared cells and free floating cells without benzalkonium and the differences in the staining intensity in individual cells were more marked. Observations revealed that the reaction intensity changes closely corelated with the stage of mitotic cycle of each cell. Namely, DPNH-D activity of the tumor cells, which generally hihger than that of leucocytes, increased remarkably in the end stage of interphase and decreases abruptly in mitotic stage reaching the lowest level in metaphase. After the metaphse the activity increased slightly and it is kept at almost the same level during the first half of interphase. This enzyme is localized mainly in the granules of the eytoplasm. The activity of TPNH-D showed the similar localizations as those of DPNH-D, though the reaction intensity is lower than that of DPNH-D. The activity of SDH of the tmnor cells is lower than that of leucocytes and its diformazan granules are localized in mitochondria. Its activity decreases in the mitotic stage the lowest level in metaphase and in the followed interphase it is kept in a almost constant low level. α-GDH activity of the tumor is lower than that of SDH but show the similar localizations as the latter.

Since the classic work of Ranvier, it is well known that the mammalian striated muscle is composed of two types of muscle fibers, i. e., the red and white muscle fibers. In the previous paper1 it has been reported that the limb muscle fibers of mammals can be divided into three types from their activities of the histochemically demonstrable oxidative enzymes. Namely, the small red muscle fibers had a higher activity of oxidative enzymes, the large white muscle fibers a lower activity and the third type of muscle fibers being called "medium fiber" or "intermediate fiber" showed an intermediate activity between those of the red and white muscle fibers.

Lysis of fibrin was first recognized by MORGAGNI in 1769, observing a liquid blood in a patient of acute death, and the phenomenon was named as fibrinolysis by DASTRE in 1893. In 1937, MACFARLANE recognized in a patient after cholecystectomy that the blood clot was lysed completely in the following morning. Since then, much attention has been paid clinically on fibrinolysis and it has been said to occur in case receiving a large amount of blood transfusion, shock, cancer, obstetric diseases, hemophilia, various drug poisonings, allergic diseases, after irradiation and after the operations of lung, pancreas and prostate. In our department, also, the similar phenomenon was recognized often in association with cardiac surgery using the artificial heart-lung machine, and a difficulty in hemostasis was encountered postoperatively. We have been studying, therefore, on fibrinolysis in open heart surgery.

The changes of rat muscle fiber structure and fiber types after the reunification of the nerve and cross-innervation between the nerve to M. soleus (SOL) and M. extensor digitorum longus (EDL) were cytologically studied and the following results were obtained: 1. After the reunification of the nerve, the tendency toward grouping to a single fiber type was observed, although in normal muscle, the red, white and intermediate fibers were distributed in mosaic pattern. 2. After the cross innervation, the changes of fiber types occurred; namely, in SOL, normally composed of red and intermediate fibers, the three types of fibers appeared after the cross- innervation with the nerve to EDL, which originally was composed of the red, white and intermediate muscle fibers, and vice versa. These changes were observed not only in histochemical sections, but also in the ultrastructural level by electron microscope.

For the purpose to reveal the correlation between molecular structure and biochemical functions of cytochrome oxidase the author studied purified cytochrome oxidase by using high resolution electron microscope and biochemical methods. 1. Cytochrome oxidase was purified from the cytochrome oxidase-rich submitochondrial membrane (green membrane), obtained from beef heart mitochondria, by three different methods; modification of the method of OKUNUKI et ai., method of FOWLER et ai. and modification of the method ofJACOBS et ai. All the preparations showed a high specific activity under appropriate conditions and consisted mainly of small particles measuring approximately 80 to 90 A. in diameter. 2. The particle, measuring approximately 80 to 90 A. in diameter, took a cylindrical form measuring about 70 A. in diameter at the base and 95 A. in height in an appropriate condition. Many experimental results indicate that the particle is the smallest, fundamental unit of the active cytochrome oxidase. For this reason it was designated as the unit particle of cytochrome oxidase (abbreviated as UPCO). 3. The molecular weight of the unit particle, calculated from its volume and average density (1.24) of lipoproteins (3: 7), was about 270,000. The value was roughly twice the minimum molecular weight of 128, 000 calculated from the heme a content. Accordingly, it is considered that the unit particle contains two heme a molecule and two copper atoms. 4. It was suggested electron microscopically that the particle collected in the 22.6 S position by sucrose gradient ultracentrifugal analysis was a dimer of the unit particle of cytochrome oxidase and also that the particle collected in the 5. 7 S position was a half of the unit particle of cytochrome oxidase. 5. It was also suggested that the particle observed on the green membrane was a subunit of cytochrome oxidase, containing one heme a and one copper atom, and the unit particle of cytochrome oxidase was constituted of two of the particles observed on the green membrane. Namely, the results indicate that the molecular state of cytochrome oxidase on the green membrane apparently differs from that of the purified cytochrome oxidase.

An attempt was made to find out the nature of catalase coritained in the red cell, especially in the ghost, For this the red cell ghost isolated were washed several times with CO2-saturated water or deionized water and the catalase activity per gram protein of the ghost was estimated. It was found that despite several washings, the catalase activity/gram protein of the ghost do not decrease as compared with the activity of the original red cell solution, indicating the presence of catalase in the ghost. In the case of hypocatalasemic blood the catalase activity in the ghost shows similar behaviors as with normal blood cells. It is assumed theoretically that there are two kinds of catalase having different affinity to the red cell ghost. Namely, one that is readily released from the ghost and the other that has a strong affinity. The affinity of hypocatalasemic blood to the ghost seems to be somewhat weaker.

In the mixed tissue culture of mouse lymphocytes with addition of PHA the rate of the appearance of large and intermediate cells increases markedly, but which side of the two cell groups have reacted stronger remains obscure. In order to solve this problem, mixed cultures were conducted in such a way that only one cell group of the two would react. Namely, one cell group was exposed to C0 6).irradiation (Table 2) prior to the culture and cultured with another viable cell group (FJ test group, Table 2) to see the percentage of the appearance of large and intermediate cells. Simultaneously, the skin homograft from respective donor mouse was transplanted to each other and the survival days of each skin graft were compared. As a result it has been shown that the percentage of blastformation and the survival time of the skin transplant in each group prove to be in an inverse relation. The results of these mixed cultures indicate that the extent of blast. formation reflects significantly the difference in B-2 histocompatibility antigens.

In vitro and in vivo experiments were conducted for the purpose to determine whether or not the antitumor factor found in the regional lymph node cells of the mouse sensitized with Ehrlich tumor cells would transfer its antitumor activity to normal lymph node cells. In in vivo experiments normal lymph node cells incubated at 5°C for 60 minutes in the supernatant containing the antitumor activity have shown the antitumor activity against JTC-11 cells in mixed culture. Namely, it hs been demonstrated that the antitumor activity in the supernatant can be transferred directly to normal lymph node cells in vitro. In the in vitro experiments, the same results as in in vivo experiments were obtained. The antitumor activity against JTC-11 cells has been detected in the lymph node cells obtained on 4, 7 and 9 days after subcutaneous and intraperitoneal injections of the supernatant having antitumor activity. Next, we tried DNase and RNase treatments of the sensitized supernant to observe the transfer factor-like snbstance. The results indicate that, while the passive transfer is possible with the supernatant treated with DNase, it is not with the RNase-treated supernatant. From these findings it is assumed that the factor (in the sensitized supernatant) capable of conferring the antitumor activity is an RNA-dependent substance (or a substance closely associated with RNA) and is probably different from the antitumor factor reported in Parts 1 and 2.

As a link in a series of studies on the effects of blood constituents on the brain function by means of brain perfusion, we used four kinds of artificial blood; namely, the blood containing a low molecular dextran, one containing glutamic acid, one containing essential amino acid group and the one containing both essential amino acid group and glutamic acid. During the perfusion experiments we observed the effects of blood constituents on the function and metabolism of the perfused brain and obtained the following results. 1. When a low molecular dextran is used as the colloid osmotic pressure agent instead of hydrodextran, the amount of the blood flow in the brain is maintained roughly at a certain fixed level throughout the experiment, showing no gradual decreasing tendency. 2. When using the artificial blood supplemented with glutamic acid, EEG of the perfused brain shows an increase in the appearance rate of β32 and β33 bands, approaching closely to the pattern of EEG of unrestrained controls at arousal state. 3. In the case of the blood added with essential amino acids similar to the case using the blood with glutamic acid, EEG approaches towards the alert pattern of the controls. 4. When the perfusion is done with the artificial blood lacking in amino acids, about one hour after the start of the perfusion the amount of glutamic acid and its related compounds in the brain can no longer be maintained at normal level and the decrease, being so marked, brings about a marked decrease also in total amino acid content. 5. When the perfusion blood contains glutamic acid, essential amino acid group or both, the concentrations of amino acids of the brain glutamic acid group and the total amino acid can be maintained approximately at normal level for the duration of over one hour.

1. The forms of irritation causing inflammation and pain are reviewed, with reference to the significance of histamine, serotonin and bradykinin and in particular to the interrelationship between inflammation and pain. 2. The various types of experimental pain are reviewed and mention is made of the human and animal analgesia test methods derived from them. 3. More detailed descriptions are given of the analgesia test methods used by us, namely: a) Silver nitrate gonarthritis-pain, rat, in which both strong and weak analgesics with an anti-inflammatory action are effective. b) Phenylquinone-induced abdominal pain, mouse, in which all the analgesics and anti inflammatory agents mentioned in this article are effective in a greater or lesser degree. c) Tail-flick and hot-plate tests, mouse, in which the strong analgesics, the weaker analgesics and the anti-inflammatory agents, with the exception of the salicylates, are effective. d) Dental-pain test, guinea pig, which can be used to demonstrate the activity of the various analgesics, including the salicylates and also colchicine, which is not active in any other test. e) Pressure-pain, mouse, in which only the strong analgesics (narcotics) are effective. 4. The action of a large number of analgesics, anti-inflammatory agents and related drugs in the various analgesia-tests and in acute experimental inflammation is presented in tabular form. 5. It is concluded that the use of several pain and inflammation tests is essential for screening both analgesics for special indications (severe, mild pain, pain due to inflammation, etc.) and universal pain-killing drugs.

The liver glucuronyl transferase (GT) activity and uridine diphosphate glucuronic acid (UDPGA) content in the patients with viral hepatitis were determined using 4-methyl umbelliferone (4-MU) as a glucuronide receptor. The results were as follows: 1. In acute viral hepatitis, the decrease in the GT activity was more remarkable in the later stage of the recovery. In chronic viral hepatitis, the GT activity was decreased in accordance with the increase in the degree of liver injury. Liver UDPGA content was significantly reduced only in postnecrotic cirrhosis. 2. The decrease or injury in the parenchymal liver cells caused a decrease in the liver GT activity. These quantitative reductions in the liver parenchyme were not the only factor for the alteration in the GT activity of the liver. The results of the present study suggested an involvement of a qualitative change in the liver GT activity in human liver injuries, especially in the early stage of acute viral hepatitis; namely, there might be even an activation of the liver GT other than the reduction resulting from the decrease in the liver parenchyme. 3. The decrease in the liver GT activity correlated significantly with the decrease in the salicylamide glucuronide formation in vivo, while the alteration in the liver UDPGA content failed to correlate with that in the glucuronide formation in vivo. It was suggested that the velocity of in vivo UDPGA production rather than the UDPGA content of the liver was as important a rate-limiting factor for the glucuronide formation in vivo as the liver GT activity.

By far the majority of studies and speculations on metabolism in man and animals have been concerned with the fate of proteins, fats, and carbohydrates. Even when interest was awakened in enzymes and vitamins, the transformation of the organic substances was still studied. Gradually, however, various methods of the analysis and measuring instruments used have been so perfected that other problems can now be included in the field of biological and medical researches, namely, the significance of inorganic substances for the living organism.