The inner membrane isolated from strain TK-4958 of Streptococcus pyogenes (Str .pyogenes) was incubated with benzyl [¹⁴C] penicillin ([¹⁴C]-PCG) at 30°C for 10 min. Then, penicillin-binding proteins (PBPs) were fractionated by dodecylsulphate-polyacrylamide gel electrophoresis, after solubilisation of the membrane by the addition of sodium lauroyl sarcosinate and non-radioactive benzylpenicillin (PCG). The properties of PBP _s isolated from Str. pyogeneswere compared with those of PBPs of E. coli and the affinities of various β-lactam antibiotics were determined with their competition for the binding of [¹⁴C]-PCG to each of the PBPs. The results obtained are as follows:
(1) Six PBP _s, PBP-1 A (mol. wt.: 96, 000), PBP-1 Bs (mol.wt.: 80, 000, 78, 000), PBP-3 (mol.wt.: 69, 000), PBP-4 (mol.wt.: 52, 000), PBP-5 (mol.wt.: 45, 000) and PBP-6 (mo.wt.: 40, 000), were identified in Str.pyogenes and this species was not estimated to have the same PBP as the PBP-2 of E.coli.
(2) These PBPs from Str. pyogenes were more heat labile than the PBPs of E. coli. E specially PBP-1 Bs was almost completely inactivated at 50°C for 10 min., and the property of [¹⁴ C]-PCG binding was abolished.
(3) The [¹⁴C]-PCG bound to PBP-5 or -6 of E. coli was easily released, while almost no release of [¹⁴ C]-PCG from the PBPs of S tr. pyogenes was observed. From this fact, the PBP-5 and -6 of this species were suggested to have no penicillinase activity.
(4) The binding affinities of various β-lactams for PBP-1 A, -1 Bs and -3, which are considered to be most closely related to lysis of bacteria, were compared by the 50% inhibitive concentration (molar ratio) of [¹⁴ C]-PCG binding by competition of β-lactams for [¹⁴ C]-PCG to these PBPs or by kinetics of β-lactams. Cephaloridine and PCG had the highest affinities to these PBPs followed by such cephalosporins as cefotiam, cephalothin and cefazolin, and ampicillin. However, there was difference in either Km value or Vmax/Km value. Piperacillin, cloxacillin and cefmetazole had lower affinities for the PBPs than the above mentioned another antibiotics, and mecillinam had almost no affinity.
From these results, it was suggested that β-lactams, which have high affinities for PBPs of gram-negative bacilli, did not always show high affinities for those of gram-positive cocci.

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Morphological changes of Streptococcus pyogenes (Str.pyogenes) induced by β-lactam antibiotics were observed under a phase-contrast microscope and an electron microscope. Relationship between the results obtained in this experiments and the affinities of the β-lactams for penicillin-binding proteins (PBPs) which had already been reported in the Part 1, was studied.
(1) By the treatment with β-lactams, the cells showed bacteriolysis at over minimum inhibitory concentrations (MICs) and the formation of football-shaped or rod-shaped cells at under MICs depending on antibiotics.
(2) Incomplete septa formation was found in several places of the cell wall in football-shaped cells with an electron microscope.From that, it was clarified that abnormal divisions of cells was occuring.
(3) From the above findings, it was suggested that such morphological changes were resulted from the affinity balances of the antibiotics for PBP-1Bs and PBP-3.That is, footballshaped cells appeared remarkably by the treatment with piperacillin or cloxacillin which showed a higher affinity for PBP-3 than for PBP-1Bs, whereas they were relatively less with cephalothin or cefmetazole which showed higher affinity for PBP-1Bs than for PBP-3.In case of other antibiotics, degree of morphological changes of cells was intermediate of them.

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The effect of hypophysectomy on the cellular immunological function has been studied in the female rats of Wister strain.They were divided into two groups, that is, hypophysectomizedand sham-operated rats.The sham-operated rats were were used as the controls.The resultsobtained were as follows.
(1) The increase in body weight was much retarded in the hypophysectomized rat, beingabout a half or less than a half of that in the control at 4-5 weeks after the operation.
(2) The weight of the thymus in the hypophysectomized rat at 4-5 weeks after theoperation was significantly lighter, as compared with that in the control.
(3) The mitogenic (Con A) proliferation of the thymus and lymphnode cells was studiedby means of 3H-thymidine incorporation.No significant difference in the proliferation wasobserved between the hypophysectomized rat and the control at 45 weeks after the operation.But the proliforation was remarkably suppressed in the hypophysectomized rat at 10-47 weeksafter the operation, as compared with that in the control.
(4) The delayed type allergic skin reaction was studied by means of immunizing the ratswith the emulsion of BSA and complete Freund adjuvant.The reactions in the hypophysectomizedrat immunized at 5 weeks after the operation were the same in severity as those in the control.But when immunized at 15 weeks after the operation, the hypophysectomized rat did not showthe reactions whereas the control did exhibit them.
(5) The influence of the sera upon the mitogenic (Con A) proliferation was studied.Thesera used were taken at 4-5, 10-42 and 15-47 weeks after the operation, respectively.Noinfluence was observed in the case of the hypophysectomized rat, as well as in the case of thecontrol.

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It has been established that the lateral cervical nucleus (LCN) in the cat is located in the first two cervical segments (C1 and C2).But in the rat even the existence of the LCN is stillcontroversial. In the present investigation, the entire cervical segments were sectioned seriallyand stained with cresyl violet in order to study the morphological characteristics of individualnerve cells.The projections to and from these nerve cells were then studied by employing the Nauta method and the HRP technique.Groups of nerve cells were found in the region ventral tothe dorsal horn in the lateral funiculus in each cervical segment.They were especially prominentat the levels of C1 and C2.These nerve cells received fibers ascending in the spinal cordhomolaterally and projected their axons into the brain stem contralaterally.These findings indicatedthat the groups of the nerve cells in C1 and C2 in the rat corresponded to the nerve cells inthe LCN in the cat.Furthermore, in the rat the nerve cells coinciding in the morphologicalcharacteristics with those in the LCN were found also in other cervical segments.

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Sera from 834 persons aged 0-68 years including cord bloods were collected in 1977-1979 at different areas in Thailand by Society for Medical Research in South East Asia, Nippon Medical S chool. These sera were examined by micro complement-fixatio (CF) test for antibodies to cytomegalovirus (CMV) and by micro neutralization (NT) test for antibodies to herpes simplex virus (HSV) type1 & 2. However, NT antibody titers were analyzed by using the II/I index in order to determine the type of HSV antibodies, because there is cross-reaction between type1 & 2 viruses.
(1) Age related CMV antibody positive persentages were79. 2% of all samples, 91.1%incord bloods, 46.7%at the age of 7-11months old, 74.1%of7-8years old, 75.3-85.9% up to 30 years old, 95.5% of older than 31years.
(2) Comparing the age related CMV antibody positive rates of different areas, it was recognized that all sera from the5-63years old of a hill tribe villege had the positive antibodies, and the antibody positive rate apparently decreased in accordance with the advance of urbaniza.-tion of the areas especially among adults. Sera with high titers of the antibody were found in all age groups in Bangkok, which suggested dense infection within small groups of low socio-economic status.
(3) In 33 paired serum samples from mother and newborn (cord blood), CMV antibody status corresponded well, each other. However, the antibody positive rate in mothers was higher than that of average adults. This suggested that latently infected CMV in the female was reactivated during pregnancy.
(4) Analysis with II/I index showed that HSV antibody was contained in 62.7% of all samples, 87.5% in cord bloods, 5.9% at the age of0-6 months old, 52.6%of 3-4 years old, 52.6-63.2%up to 20 years old, and 70.8%of 21-25years old.
Increase of HSV seronegative young adults has recently attrated attention in the developed countries, but this tendancy was not yet recognized in Thailand.
(5) By using the II/I index, NT antibody to type1was found in about10%of children below 2 years old and about 50% of 3-10 years old. On the contrary, type 2 antibody was found in 10% of 3-20 years, and 18% of over 21 years old.
(6) Although antibody status by age was somewhat different in different geographical areas, primary infection of HSV was supposed to occur in higher ages in Bangkok compared to another rural areas.
(7) Thirty-three pairs of sera from mother and newborn were analysed with II/I index. Type1antibody was found in both samples of 21 pairs, and4pairs were antibody negative. In 8pairs, mother and newborn antibodies were not identified as same type, but the difference of NT antibody titers between mother and newborn were less than 2 times, except 2 pairs.

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Thickened or ossified ligamentum flavurn are taking interested in orthopedic surgeon srecently, because of much of these patients are liaiable to complain the spinal cord or cauda equina compression syndrome. The author performed to investigate the postnatal development and these changed ligamentum flavum with aging by light microscopically and electron micro-scopically.
Besides histochemical localization in the ligamentum flavum was examined by means of Alkaline phosphatase, Acid phosphatase, and Adenosine triphosphatase as well as collagenase digestion and elastase digestion methods.
Furthermore yellow pigment of the ligamentum flavum was identified with thin layer chromatography.
Results: 1) In a newborn stage of the ligamentum flavum, specimens showed much collagenous fibrils, stained strongly positive by PAM, and sparse elastic fibers.
2) The elastic fibers in a newborn stage appeared as an aggregation of PAM-positive microgranules mainly in PAM-negative collagenous matrix and in partly collagenous fibrils. It was clearly defined these microgranules in initial stages as PAM-positive amorphous material after elastase digestion. It was noticed that the PAM stain was most adequate method for staining these elastic fibers in a very initial stage, compared with other staining methods.
3) The elastic fibers contained PAM-positive microfibrillar component in its interior, which was thought to be deformed collagenous fibrils in stages from newborn to adolescent. Wheas, that in a stage of adult or later did not contain those fillamentous material, but electron dense granular substance.
4) In the matured elastic fibers, the author found clearly the microfibrillar layer in periphery of them. It was confirmed that the microfibrillar layers consisted of microfibrils as well as amorphous material, should be called “microfibrillar ground substance.”
5) With double staining method after collagenase digestion, the author found the microfibrils clearly defined and the microfibrillar ground substance widened. With PAM stain after this digestion, PAM-positive materials disappeared in elastic fibers.
6) With using double staining method after elastase digestion, vesiculation was observed in elastic fibers of an infantile stage. In elastic fibers in an adult stage, the enhancement of interior fillamentous or reticular materials was noticed, but the microfibrillar layers were not digested by elastase. The microgranules, stained strongly positive by PAM stain, remained unchanged in PAM stain after the digestion.
7) Based on these data, it is presumably apparent that the elastase digests neither the elastic fibers completely nor microfibrillar layers, but does main elements partially. Opposed to asumption, it is realized that the collagenase does not give any significant alterations, but does the elastic fibers some changes to a certain extent, as far as objected by PAM stain.
8) It was found that the yellow pigments of the ligamentum flavum may be equivalent to riboflavin and flavin mononucleotide (FMN).

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Success of a surgical operation largely depends on the pre-and post-operative nuturition control. The tube feeding has several advantages over other methods such as postoperativeperipheral venous infusion or intravenous hyperalimentation. On the other hand, tube feedingis not free of side effects. The most serious of the side effects is diarrhea. On the assumptionthat diarrhea resulting from tube feeding is caused by lactose intolerance, which is foundamong a considerable proportion of the Japanese people, the author has made a detailed studyon the following points in the postoperative cases of the upper GI tract disorders:
(1) The proportion of milk intolerance.
(2) Lactose tolerance test.
(3) Measurement of disaccharidase in the mucosa of the small intestine.
(4) The clinical effect of lactase preparation.
The results are as follows:
(1) Average ratio of the disaccharidases in the mucosa of the intestine was:
Lactase1: Sucrase: Maltase=1:11.7:52.8
The ratio of lastase 1 is one fifth to one sixth of that found in the Westerners.
(2) In the present study, a'lactose intolerance case'refers to a subject who had less than20mg/dl in the 25g lactose tolerance test or less than 10u/g prot. in lactase 1 activity value.
(3) When lactase preparation was administered, the incidence of side effects resulting fromtube feeding the liquid diet containing lactose was6%, in contradistinction to the incidence of70. 6% which would have otherwise occured.
(4) More than 1g of lactase preparation was required for each 5g of lactose for patientshaving milk intolerance. Usually, however, 1g of lactase preparation was sufficient for each7to 8g of lactose.

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This report demonstrates a quantitative method of measuring host defense against syngeneictransplantable solid tumor (mammary adenocarcinoma 755 raised in C 57 BL/6 female mice). Three preparation methods, solid tumor mass, screen-meshed tumor cells and trypsin treatedtumor cells, were investigated. In these methods screen-meshed tumor cells were easilyobtained and fairly quantitative. Since tumor cells were inoculated into the left hind footpadof syngeneic mice, the kinetics of tumor growth during the early stage (from the time of tumorcells inoculation to that of footpad reaching about 3. 6 mm) were simply expressed by the twoparameters of latent period and rate of increase. Some factors affecting these parameters oftumor growth were next investigated. Sex differences were observed. The latent period wasaccerelated in female mice at 24 weeks old, but the rate of increase in both sexes was notdifferent. Aging in male mice, from 5 to 60 weeks, affected the latent period which wasgradually prolonged with aging, but the latent period in female mice was not affected by age. No correlation between the rate of increase and aging was not observed in either sex. Doseof dead tumor cells did not affected tumor growth. Latent periods were significantly prolongedin mice after thymectomy and 600 R whole body irradiation as compared to that of untreatedmice, but were somewhat shortening in mice receiving thymus cells after thymectomy andirradiation.
On the other hand the rate of increase in untreated mice and the mice receiving more than1×10⁷ thymus cells were lower than in irradiated mice and the mice receiving fewer than 1×10 ⁷ thymus cells. Bone marrow cells significantly prolonged the latent period, in proportion tothe dose.
Latent period in mice treated with antimacrophage agent (ferritin) was significantlyaccelerated.

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