It has been proven that demethylchlortetracycline (DMC) inhibits ADH-responsiveness and aldosterone-mediated Na⁺ transport in toad urinary bladders. There are reports that DMC can cause nephrogenic diabetes insipidus (DI) in man. But whether DMC has natriuretic action or antialdosterone effect is unclear. Most of the studies of DMC have been limited to in vitro or clinical. In clinical studies, the actions of nephrogenic DI has been found by accident. This author studied the diuretic action of DMC in rats and tried to find whether DMC has natriuretic action in rats. The author also studied DMC as a diuretic in patients with various disorders to try to find if DMC has diuretic and natriuretic action. Experiment 1: DMC, in the dose of 20 mg/kg of body weight (BW), was given intraperitonealy for 14 days, and a control group received normal saline, with daily measurements of urine output, osmolality, and urinary cyclic AMP (cAMP) in both groups. A 40-hour urine concentration test was performed on the 10th dey. On the 13th day, aldosterone was administered subcutaneously. Urine output was found to be greater, and urine osmolality and cAMP were lower, in the group receiving DMC. On both the 10th and 13th day Na/K ratio was higher than in the control group (sig. diff. p<0.001), proving the anti-aldosterone effect of DMC. Experiment 2: Two groups received DMC intravenously (30 and 60mg/kg BW), and a control group received normal saline. The group reveiving 60mg/kg BW of DMC showed significant increase of urine output and significant decrease of urinary cAMP and urine osmolality. But these changes were not observed in the group which received 30 mg/kg BW of DMC. It was suggested that DMC produces reversible ADH unresponsiveness by inhibition of both cAMP genepation and action. The previous two experiments indicated higher levels of magnesium (Mg) excreted in the DMC group, although the exact mechanism is unknown. Experiment 3: 900 to 1200mg of DMC was administered orally to 13 patients with diagnoses of idiopathic edema, cirrhosis of the liver, nephrotic syndrome, chronic glomerulonephritis and cerebrovasculr accidnet.10 patients showed water diuresis and three did not. Four showed natriuresis and 9 did not.4 patients who received 1200mg of DMC daily were found to have decreases in urine osmolality and urine cAMP, which were reversible. A typical case which showed remarkable diuresis with D MC was described. It is suggested. that DMC is useful not only for the treatment of hyponatremia but also for the edematous status which accompanies it.

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Determinations were made with Campbell's method of serum 5 '-nucleotidase (5'-N) activity in 116 cases of liver, gallbladder diseases, gastrc cancer and bone diseases. At the same time activities were studies of transaminase (TA), isocitric dehydrogenase (ICD), alkaline phosphatase (ALP), leucine aminopeptidase (LAP) and β-glucuronidase (β-G). Also, S. D. strain rats subjected to acute CCl₄ hepatic disturbances, common bile duct ligation and DAB hepatoma were determined as to their serum 5'-N, ALP, LAP and β-G in an attempt to evaluate diagnostic value on these diseases with the following results:
1) Twenty healthy adults showed their 5'-N activity to be 6.5 ± 4. Ou.
2) Cases of acute viral hepatitis showed only a slight increase in 5'-N, while those with erythromycin estolate induced intrahepatic cholestasis showed a marked increase which proved to be were useful in evaluating challenge tests than ALP and LAP.5 '-N was favorable when used as an index for intrahepatic cholestasis.
3) Cases of inactive type of ch ronic hepatitis showed only a very slight increase in 5'-N.
4) Cases of liver cirrhosis showed a slight increase in 5: those of noncompensato ry type showed higher increase than a compensatory type, revealing an abnormal rate of 80% with their higher 5'-N than ALP.
5) Cases with p rimary liver cancer showed a slight increase and those with metastatic liver cancer a moderate increase in 5'-N, both accounting for a 100% abnormality.
6) Cases with cholelithiasis at remission showed normal 5'-N. But those with cholelithiasis 10-14 days after onset showed higher 5'-N than ALP and LAP, revealing a significant diagnostic value.
7) All cases of gastric cancer without metastasis to the liver and other showed normal enzyme activity inclusive of 5 '-N.
8) Regarding bone dseases, one case with diffuse bone metastasiis and one case of bone metastasis whose determination was made immediately before death showed a normal range of 5 '-N and ALP. Bone metastasis cases of gastric cancer and cases of bone fracture showed an increase in ALP in 87.5%, while their 5'-N was kept normal. Thus, it was considered necessary to make a simultaneous determination of 5'-N and ALP in differentiating cases of osseous ALP and liver ALP.
9) In chronological observations rats with acute CCl₄ hepatic disturbances and those of common bile duct ligation showed the highest increase in 5'-N in three days, especially in latter rats. Rats with DAB hepatoma showed a slight increase in 5 '-N, while their ALP and LAP remained normal. Thus, it was demonstrated that 5'-N in rats of common bile duct ligation and those of DAB hepatoma reflected changes in enzyme activity on a clinical basis.

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Patients with acute inferior myocardial infarction (MI) are sometimes associated with right ventricular (RV) infarction and they often develope a characteristic clinical feature with disproportionate elevation of RV filling pressure compared with left ventricular filling pressure. In this study, RV function in patients with RV involvement was investigated with a special reference to regional wall motion abnormality of the right ventricle. Fifty-nine patients with old MI and 18 cases withou t any apparant cardiac abnormalities (normal group) were studied by the first-pass radionuclide angiocardiography. The global function of the right and left ventricles (RV and LV) were estimated as ejection fractions, RVEF and LVEF, respectively. Regional abnormality of RV myocardial wall was determined on a regional ejection fraction image, which was computed as a stroke-volume image divided by an end-diastolic image of the right ventricle. Among 34 cases with inferior MI,10 patients (29.4 %) had been diagnosed clinically in their acute phase as having RV infarction. Then, MI patients studied were divided into 3 groups: 25 cases with anterior MI,24 with inferior MI without RV involvement and 10 cases with RV infarction.
All the patients with RV infarction had a depressed regional ejection fraction area at the lower part of RV. Patients with MI showed significantly lower LVEF than the normal cases. Anterior MI group had the lowest LVEF value among the groups. Inferior MI and RV infarction groups showed lower RVEF than anterior MI and normal groups. RV infarction group had the lowest RVEF value among the groups. RVEF/LVEF ratio was significantly higher in anterior MI group than in the other groups. The regional ejection fraction value of the inferior part of RV (RVinf) was significantly depressed in inferior MI and RV infarction groups. The anterior part of RV (RVant) showed significantly depressed ejection fraction value in anterior MI and RV infarction groups as compared to the normal group. Thus, RVinf/RVant ratio was significantly lower in RV infarction group. RVinf/RVEF ratio was also significantly lower in RV infarction group. There was no significant difference in RVant/RVEF ratio among th e groups. In summary, in patients with RV infarction, global right ventricular function was substantially impaired due to depressed wall motion of its lower part. Accordingly, lower ejection fraction at the inferior part of the right ventricle as compared to the anterior part and the whole right ventricle, namely, lower values of RVinf, RVinf/RVant and RVinf/RVEF, would be useful hallmarks for diagnosing RV involvement associated with inferior LV infarction. Moreover, even in the absence of the signs and symptoms suggesting RV infarction, the regional wall motion abnormality in the lower part of the right ventricle, if demonstrated in the clinical setting of inferior infarction, should indicate the right ventricular involvement.

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Isolated sinus node of rat heart was fixed on a cork board immersed in Locke's solution saturated with oxygen at 30°C, and the frequency of its spontaneous activity was counted by means of Brown oscillograph. N-ethylmaleimide (NEM) and parachloromercuribenzoic acid (PCMB), which are SH-reagents, suppressed the frequency of sinus node (SNF) in their resp ective adequate doses. Trimetazidine (TMZ), also, decreased the SNF, but when the drug was present in the medium, the drug inhibited the effect of NEM and PCMB. L-cysteine, which has an SH-radical in its molecule, also, inhibited the effect of NEM and PCMB. It was an interesting result that TMZ which suppresses the SNF inhibited the action of NEM or PCMB which suppresses the SNF.
In order to exclude the doubt whether the chemical reaction between TMZ and NEM or PCMB might cause the decrease of biologically effective dose of these SR-reagents or not, chemical reactions between NEM, PCMB, TMZ and L-cysteine were examined by means of UV-absorption spectrum method and thin layer chromatography. Between NEM and L-cysteine, and between PCMB and L-cysteine, chemical interactions were seen, and the former was pH-dependent while the latter was not pH-dependent. Although TMZ did not cause chemical reactions with either NEM, PCMB or L-cysteine, TMZ promoted the chemical interactions between L-cysteine and NEM or PCMB. Then, it was assumed that the effect of TMZ to inhibit the suppressive effect of NEM and PCMB on SNF is not chemical but biological one.
The effect of NEM on SNF reduced when K⁺ in Locke's solution was decreased to 1/ 2 K⁺or when Ca²⁺ was increased to 2 Ca²⁺, and the effect of PCMB on SNF augmented when Na+was decreased to 1/3 Na⁺ or when K⁺ was decreased to 1/2 K. The effect of NEM on SNF disappeared after the drug was removed from⁺ the medium, while the effect of PCMB remained even after its removal from the medium.
From the results, the author a ssumed that NEM may affect the K⁺-channel by occupying SH-radical following the promotion of K⁺-flux and inhibition of Ca²⁺-flux through membrane, that PCMB may affect the Na⁺-channel by occupying SH-radical following the inhibition of Nat-flux, and that the effect of NEM which is water soluble was reversible while that of PCMB which is oil-soluble was irreversible. The effect of TMZ was assumed to be partially due to inhibition of K⁺-flux and partially due to inhibition of Ca²⁺-flux.

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In the first of serial studies carried out on lymphocyte function in gynecological patients, the correlation between lymphocyte reactivity against mitogens (PHA, Con A, and PWM) and prognosis or clinical staging were examined to yield the following results:
1) The T-lymphocyte percentage was slightly reduced in concomitance with the developmental stages in uterine and ovarian cancer, the most remarkable decrease being obs erved during stage IV.
2) In uterine cancer patients, the lymphocyte reactivity to PHA was reduced in concomitance with the disease stages, while the reactions to Con A and PWM remained no rmal during stages 0 through II. In ovarian cancer patients, the reactivity against all three mitogens was concomitant with the developmental stage, but the reactivity to PHA was found to recover after tumor removal.
3) In both uterine and ovarian cancer, a remarkable lowering in these mitogen reactivities was seen about 2 months before death.
4) The sera in progressed cancer cases showed the inhibitory effects of reaction against mitogens.

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In the second of these three serial studies the effects of therapeutic Co⁶⁰ irradiation on the lymphocytes were examined with special refrence to their reactivity against mitogens and antilymphocyte antibody to yield the following results:
1) Total lymphocyte counts in the peripheral blood and the absolute T cell number decreased after irradiation, although no changes were found in T cell percentage.
2) In order to evaluate immune response in uterine cancer patients undergoing therapeutic irradiation, in vitro lymphocyte transformation tests using PHA, Con A, PWM and PPD, as well as mensuration of IgG, IGA, IgM, C₃, and C₄, were carried out on the blood obtaine d from each patient before and during therapy: During irradiation, the lymphocyte trans for-ma tion values against PHA and Con A were depressed, but no significant changes were found in the values of IgG, IgA, and IgM. In some patients, however, the IgG, IgM, C₃ an d C₄ values were slightly increased after irradiation.
3) Using sera with high anti-lymphocyte antibody titers obtained from SLE patients, a cytotoxicity test against normal lymphocytes and lymphocytes from Co⁶⁰ -treated patients was carried out: In approptiate seral dilution, less cytotoxicity was found in lymphocytes obtai ned from Co⁶⁰ treated patients than in those obtained from normal patients lymphocytes, i.e. lymphocytes from Co⁶⁰ treated patents were found to be more resistant to the cytotoxic sera than normal lymphocytes.

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In the third of these three serial studies, the natural killer (NK) activity in cancer patients, which may play an importnat role in the immune surveillcnce against cancer was examined to yield the following result:
1) Target cell, K 562 or Molt-4, were added to effector cells (mononuclear cells separated from the peripheral blood), and the NK activity of the latter was evaluated by th e dyeexclusion technique. K 562 showed a higher susceptibility than Molt-4, but a distinct corr elation was found between the susceptibity of both cell kinds. An increased ratio in the number of effector cells to that of target cells bought about an elevation in NK activity. Incubation tim e (4,6, and 20 hr) presented no considerable influence on the data.
2) The NK activity fell during the late cancer stages. Otherwise, the uterine cancer patients continued to show an almost normal NK activity, while the ovarian cancer patients ten ded to show a generally lowered activity both both before and after operation. Co⁶⁰ irradiation exerted no influence on NK activity in uterine cancer patients. No cerrelation could be found between NK activity and T-cell percentage.

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Alkaline phosphatase activity (AP) and acid phosphatase activity (SP) were histochemically studied in the neutrophils of healthy individua ls and in those of patients with blood disorders, i.e.acute granulocytic leukemia (AGL), chronic granulocytic leukemia (CGL), m yeloma, aplastic anemia (APA), idiopathic thrombocytopenic purpura (ITP), and essential hypochromie (HA). Neutrophils obtained from the peripheral blood or from the hone marrow by puncture were stained by the azo-dye method.
The results obtained were as follows:
1) Immature neutrophils from the bone mar row of normai individuals showed a lower AP and a higher SP than mature neutrophils. Comparing these activities in mature neutrophils from the bone marrow with those from the peripheral blood, the former showed a lower AP and a higher SP thain the latter.
2) In AGL, the AP varied widely, being high in the non-remissive phase and approachiog normal during remission.
3) CGL patients showed a low AP and a high SP in both the peripheral blood and bone marrow neutrophils.
4) In the bone marrow of myelome, the AP of the mature neutrophils was high, while the SP was similar to the control level.
5) In APA, the AP was high both in the peripheral blood and bone marrow neutrophils, whils the SP was normal in the immature neutrophils and high in the mature ones.
6) ITP and HA patients, the neutrophilic AP and SP were similar to those of the normal controls.

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With a view to analyzing the inhibiting effect of anticancer drugs and irradiation on hematopoiesis in rabbits neutrophil (pseudoeosinophil) counts and the neutrophilic activities of alkaline phosphatase (AP) and acid phosphatase (SP) were serially followed up after drug administration or irradiation. The enzym activity was estimated histochemically, using azo-dye staining. Each rabbit was given cyclophosphamid (CP) (25mg/kg × 10,at intervals of 5-7 days; 50mg/kg × 5, every day; or 100mg/kg×1,i.m.), Thio-TEPA (4mg/kg×1,i.m.), Vinblastin (VBT) (1mg/kgx1,i.v.),6MP (25mg/kg×1,p.o.), or Mitomycin C (MMC) (1.5mg/kgx1,i.v.). The results obtained were as follows:
1) The neutrophil counts became slightly elevated at 24 hrs, reached their nadir at 48 to72hrs, and recovered to normal in 5 to 6 days thereafter, except with 6 MP which produced no significant change but for a temporary elevation after dosages.
2) Except in the group administrated 6MP, which caused no significant hematorogical changes, the AP changes were similar in all of the animal groups: after temporary depre s s ion, it became elevated for 5 to 6 days, and recovered to normal about 9 days thereafter.
3) SP showed no changes in the 25mg/kg×10 CP and the 6MP groups, it became elevated in 2 or 3 days after the administration of MMC, VBT, or Thio-TEPA to recover to normal in 5 to 10 days thereafter.
4)⁶⁰Co irradiation (1,000 rad/whole body×1) led to a temporary ascent in phil count followed by a descent from the 6th day on, and then a slow recovery to normal. AP was elevated from the third to the sixth days, and, after a depression on the tenth day, it returned to normal 24 days after irradiation, while SP showed a continued elevation from the 2nd to the 13th day.
Regarding AP as a param eter for the maturation of neutrophils, and SP as a parameter for their degeneration, the present results may serve to evaluate certain drugs and irradiation on the basis of their effects on hematopoietic function.

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It has bog been pointed out that many blood dialysis patints suffer from immunologioal Pificiency. However, the reports on its context do not always agree. In fact, there are m any problems in the day dealinge with dialysis patients such as a high tuberculosis controtion r ate, many toxins infection cases, fast development in cases of infection and a high rate of malig n ant tumor outbreak. This fndicates the reduced functions of the host defence mechanism against infection. The main cause of this will be the reduced immunological response of the hos t. For this reason, the author examined various kinds of immunity factors, especiailly cell mediated im munity, in order to study the immunity conditions of ohronis cases.
(Object and Method of Stndy)
About 50 cases of chron i c renal failure who had undergone blood dialysis for a period of time were chosen as the subjects. Five to eight healthy peopie were also used to compare the results.
Following tests were undertaken;
1) Measurement of lymphocytes s ubpopulation by cell surface marker.
2) Lymphocyte blastogenesis of peripheral blood by mitogen. (PHA)
3) Target cell lesion test. Anti dependent cel l m ediated cytotoxicity. (ADCC)
4) Delayed type hypersensitivity skin reaction. a) Delayed type hypersensity react ion by recall antigen. (PPD)b) Delayed type hypersensitivity reaction by DNCB.
5) Non-specific skin reaction. (PHA skin reaction)
(Results and Discussion)
The number o flymphoytes of the patients were 1343 ± 564/mm³ which was consideralby lower than that of the healthy subjects 2494 ± 271/mm³. (P <0.001) As for the distribution of the lymphocytes subpopulation, T cell percentage was high and B cell percentage was low.
Lymphocyte blastogenesis by PHA tended to be lower with the patients' group.
The K cell ratio was markedly lower than that of the healthy subjects, but no relations were observed with the period of dialysis. The delayed type hypersensitivity reaction by PPD was low with dialysis patients, whioh indicates the weakening of theif oellular immunity ability. Also, once or twice a year of continuous tests proved the increase of reaction each year.
The result of DNCB sensitization clearly showed the reduced cellular immunity abilyty of diality of dialysis patients. The chance of natural recurrence was obviously higher among the patients who had undergone the dialysis over five years. This suggests the possibilit yof immunity improvement by dialysis.
The PHA hypo dermic test proves generally low among the patients. However, detailed study of the cases shows that although the decrease after two or three yesrs of dialysis, the tendancy increases again with the cases of experience with dialysis over five years. The results of the PHA underskin tests showed a certain degree of relationship with DNCB reaction and PPD reaction.
(Conclusion) The results of thg above-mentinoed tests show that the cellular lmmunity of chronic dialysis patients is considerably low at the initial stage of dialysis introduction; however, it improves to a certain degree after five years of treatment. This suggests the possibility of immunity improvement due to the eradication of uremic toxins by dialsis on the assumption that uremic toxins are the cause for cellular immunnity decrease with chronic dialysis patients.

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