The leucocyte phagocytosis of ³H-BCG was determined in vitro using peripheral blood taken from patients of pulmonary tuberculosis both with and without complication of diabetes mellitus, and the results were analyzed in relation to humoral factors. The following results were obtained: 1) Hyperglycemia (glucose>250mg/dl) did not affect the phagocytic activity, but stimulated exocytosis of bacteria which took place immediately after the phagocytosis. 2) High serum concentration of free fatty acids (>0.25mM) reduced the phagocytic activity. 3) Anti-mycobacterial antibody, essential for opsonization of BCG bacilli, was found sufficiently in all diabetic patients.
In addition to the above clinical study, diabetic humoral abnormalities were reconstituted using normal blood, and a similar experiment was carried out. This gave the following findings: 1) High glucose concentration (500mg/dl) stimulated exocytosis of bacilli, which was accompanied with leucocyte damage. 2) Saturated fatty acids (1mM) inhibited phagocytosis. 3) Ketone bodies and acidosis did not influence the phagocytic function. 4) Insulin had no direct effect on the phagocytosis.
The above observations seem to indicate that the humoral immunity is intact in diabetics but the serum abnormalities inherent to diabetes such as high blood glucose and high free fatty acids may interfere with the normal function of phagocytes which appears to be important in the effective defense against tuberculosis.

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In 1976, we reported a paper under the title of “Comparison of the effects between combined chemotherapy with and without rifampicin (RFP) in original and retreatment of pulmonary tuber culosis with special reference to the grade of improvement of chest X-ray findings” (Kekkaku, 51: 353, 1976), and concluded that the grade of improvement on chest radiogram in RFP group and non-RFP group was similar, although the improvement was seen faster in RFP group than in non-RFP group, and accordingly, a marked shortening of the duration of chemotherapy in chronic pulmonary tuber culosis could not be expected even though a regimen including RFP might be stronger than another combination without RFP.
The above conclusion was obtained from two years observation after starting treatment, and we continued follow-up for another three years (total five years), exchanging some inadequate cases for another adequate cases, and investigated whether there is any difference between the course of the RFP group and non-RFP groups.
In the present study, we added further a new group to the original treatment cases: that is, a group who started treatment with regimens not containing RFP and then seven months or later from the start of therapy RFP was used supplementarily (RFP supplemented group). Tubercle bacilli in sputum converted negative in nearly all cases, and there was no significant difference in the results among groups treated with different regimen.
On chest radiogram, a little surpassing improvement was observed in RFP group of original treat ment as compared with other groups, though background factors might be rather favourable in RFP group. This exceeding improvement of RFP group became clearer when putting together all the data obtained from “Standard for the evaluation of X-ray findings including basic lesion & cavities”, “Standard for the summarized evaluation of response to chemotherapy of pulmonary tuberculosis” and “Standard of the attainment of therapeutic targets” (Gakken).
On the contrary, the results both in RFP supplemented group and RFP retreated group was not so good as that in RFP group of original treatment.
As mentioned above, the grade of improvement was more complete in the RFP group of original treatment than in the non-RFP group of retreatment, but we could not reach a conclusion from this study whether the shortening of the duration of chemotherapy in chronic pulmonary tuberculosis is possible or not by adding RFP.

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Diaminopimelic acid (DPA) is a component of cell wall of Mycobacterium, Rhodococcus, and Nocardia. A comparative study of the uptake of radioactive DPA has revealed that these three genera have different patterns of the uptake of this compound.
Test organisms were cultivated on Ogawa egg medium at 37°C for 5 days. The bacteria grown were harvested and washed three times with a 0.9% NaCl solution. The bacteria were weighed asep tically, and 100mg, wet weight, of bacteria were suspended in 4.0ml of a M/15 phosphate buffer solu tion (pH 7.1) containing sodium acetate (2.5μg/ml) and 2, 6-diamino (G-³H) pimelic acid dihydrochloride (25μCi/ml). The radioactive DPA (³H-DPA) was a product of the Radiochemical Center, Amersham, England (specific activity, 2.6Ci/m mol; batch 40). The reaction mixture was incubated at 37°C for 24 hours. The mixture was centrifuged, and supernatant was discarded. The bacteria were washed three times with 5ml of distilled water, and fractionated according to the procedure of Schneider (J. Biol. Chem. 161: 293, 1945).
The bacteria were first extracted twice with 2ml of a 10% trichloroacetic acid (TCA) solution (Fraction I), and twice with 2ml of ethanol and then once with 3ml of a boiling ethanol-diethyl ether (1: 1) mixture for 5 minutes (Fraction II), and extracted twice with 2ml of 5% TCA solution at 90°C for 15 minutes (Fraction III). The residue was dissolved in 2ml of a 1% NaOH solution by heating at 100°C for 5 minutes (Fraction IV).
A 0.5ml sample of each fraction was added to 5ml (Fractions I to III) or 10ml (Fraction IV) of a scintillator solvent, and the radioactivity was estimated by a liquid scintillation counter (Aloka LSC 651, Nihon Musen Co., Tokyo). The scintillator solvent used was composed of the following: Toluene, 1, 000ml; 2, 5-diphenyloxazole, 4g; 1, 4-bis [2-(5phenyloxazolyl)] -benzene, 100mg (TDP-1-SL). The radioactivity was expressed as disintegrations per minute (dpm). Total radioactivity in a whole fraction was calculated by multiplying the radioactivity estimated by the volume.
The radioactivity from ³H-DPA was incorporated first principally into the Fraction II, which contains lipids, and then moved to the Fraction IV, which contains proteins (Table 1). In this study, the radioactivity in various organisms were compared at a stage, at which the majority of the radio activity was present in the Fraction II (Table 2).
Comparisons of the radioactivity in Fraction II of various organisms are shown in Table 3. Rapid ly growing mycobacteria showed a large amount of the incorporation (60, 650 to 1, 217, 900dpm), whereas rhodococci and nocardiae showed a small amount of it (2, 648 to 44, 570). An intermediate between rapidly and slowly growing mycobacteria, M. flavescens, showed a radioactivity of in between these above two groups. It is considered that the uptake of the 3H-DPA is influenced by the growth rate, as DPA is a cell wall component. Comparison of the uptake would be meaningful, only when test organisms have the same growth rate. Rapidly growing mycobacteria, rhodococci, and nocardiae grow abundantly after incubation for 3 days, and are considered to have almost the same growth rate. Difference of the uptake of ³H-DPA between the rapidly growing mycobacteria, and the rhodococci and nocardiae would be considered as some difference of cell wall formation of these organisms.
In summary, rapidly growing mycobacteria took up more amounts of 2, 6-diamino (G-³H) pimelic acid dihydrochloride than did rhodococci and nocardiae. No marked difference was observed between the latter two.

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A case of 28-year-old female patient suffered from atypical mycobacteriosis with congenital heart failure was reported. She has been already diagnosed as congenital heart failure when she was 6 month old. At the age of fourteen, she was first diagnosed as pulmonary tuberculosis and received anti tuberculous drugs for 2 years. Subsequently, abnormal shadow on the chest X-ray was found at the age of twenty one. Smear and culture of the sputum demonstrated acid fast bacilli. She was retreated as the recurrence of tuberculosis, but clinical course was not satisfactory. From further examination of her sputum, it was demonstrated that causative bacteria was classified as M. intracellulare by Dr. M. Tsukamura (National Chubu Hospital) and it was drug resistant against all the anti-tuber culous drugs.
On the other hand, cardiac catheterization revealed cardiac disease as the corrected transposition of the large vessels with ventricular septal defect and valvular pulmonary stenosis. Operative treat ment against the heart disease was performed, although in the sputum culture acid fast bacilli remained positive. Ten months after cardiac surgery, cavitary change on the chest X-ray disappeared and the cultures of sputum became negative in spite of in teruption of drug administration. It is concluded that atypical mycobacteriosis was healed with the improvement of pulmonary circulation by heart surgery, without any anti-tuberculous treatment.

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A 72-years-old male who has been a heavy smoker suffered from pulmonary tuberculosis some thirty years ago and calcified foci were seen in the left upper lobe. A tumor opacity was found in the left upper lobe, and it grew very rapidly. The left upper lobe lobectomy was performed, and the pathlogical-anatomical examinations of the resected specimen revealed the following findings:
1) The rapid increase of the tumor opacity was caused by the activation of dormant tuberculous foci most likely due to the invasion of contiguous tumor in the external side of the tumor, where certain changes might exist due to cancerous bronchial obstruction.
2) Histological type of the tumor was epidermoid carcinoma.
3) It was suspected that the early lymphogenous metastasis of cancer to the regional glands was suppressed by the existence of old glandular tuberculous changes.

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