A method was described in which the origin of FDP could be differentiated by gel chromatography. In the preliminary experiment the stabilized fibrin was digested overnight at 37°C. The resulting fibrin DP was chromatographed on Sephadex G200 column (26×450mm) and the elution volume (Ve) of fibrin DP as well as E was measured by the use of single radial immunodiffusion or antibody-coated latex aggregation test. The Ve of fibrin DP-D was calculated as 98ml and that of E was 141ml. Fibrinogen (Fbg) was digested with plasmin and resulting Fbg DP was chromatographed on the same gel column. The Ve of Fbg DP-D was 128ml and that of E was 140ml. The difference of Ve between both FDPD was explained as the dimer composition of fibrin DP-D of stabilized fibrin wheras the monomer form of fibrinogen DP.
The urine containing 10γ/ml of FDP of a case of renal failure was concentrated 20 folds and chromatographed on Biogel A-5m column, which showed that the most of the FDP was early one, as the Ve was slightly larger than that of fibrinogen. The concentrated urine was digested with plasmin and applied on Sephadex G200 column. The chromatography revealed that the resulted FDP-D was monomer, which indicated the FDP in the urine was not originated from stabilized fibrin, but from fibrinogen or non-stabilized fibrin.
The factor was examined which interfering the result of this differentiation method. Monodansylcadaverine (MDC) incorporation with F. XIII at various digestive stages of Fbg or fibrin DP was examined basing on the method by Lorand and Urayama. It was found that the early Fbg DP had a slight capability of MDC incorporation while none of fibrin DP. This indicated that the possible crosslinkage of early Fbg DP with F. XIII and if this occurred, the resulting FDP-D could be interpreted as originated from already stabilizedfibrin by the differentiation method.
A mixture of purified fibrinogen, plasminogen and urokinase was kept at 37°C and aliquots were taken at different incubation time, then thrombin-calcium was added to each aliquot. SDS-disc electrophoressis after the reduction with DTT of each aliquot revealed that, although Fbg DP-X could be crosslinked with F. XIII and formed subunit-gamma dimer, but most of it, at the presence of fibrinogen, could be incorporated into fibrin by the defibrination procedure. With these result, the interference on the differentiation method by the crosslinkage of Fbg DP was regarded as minimal.

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In disseminated intravascular coagulation syndrome, there appear thrombin, factor Xa, plasmin, and trypsin in the case with severe pancreatitis. These enzymes are called serine protease. Defibrase (snake venom, Bothrops Marajoensis) is a defibrinogenating agent.
In this paper interaction between factor XIII and serine proteases or Defibrase was studied using citrated normal human plasma and Factor XIII concentrates (Behring Werke A. G.).
Factor XIII activity was assayed by inhibition test by reagents for determinating coaulation Factor XIII (Behring Werke A. G.).
The results were as follows: In citrated plasma, factor XIII activation and fibrin polymer cross linking by addition of thrombin were observed by this study.
The decrease of factor XIII in disseminated intravascular coagulation syndrome would be due to comsumption and absorption to fibrin.

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The plasmin digestion of factor XIII in the plasma with congenital afibrinogenemia has been reported by Suzuki et al. (1967).
The present investigation is attempted to obtain some new aspects in the biochamical changes of factor XIII during the degradation with plasmin. The authors advanced the polyacrylamide disc electrophoresis and the cross-immunoelectrophoresis with anti-factor XIII (A & S) serum for the detection of molecular change of factor XIII and our modified method for the assay of factor XIII activity. In summary, it is concluded that the process of plasmin digestion of factor XIII can be divied into three steps described below.
The first step; the subunit A of factor XIII such as placental factor XIII is ready to get plasmin digestion following the decrease of amount and activity of subunit A.
The second step; in the observation of the subunit A₂S complex like plasma factor XIII, the subunit S portion in the complex is more readily affected by. the plasmin digestion than the subunit A of the complex. Therefore, it is assumed that the subunit S may be play a protective role against the plasmin attack on the subunit A of the subunit A₂S complex.
The third step; subunit A₂S complex coexisting with fibrinogen or plasma protein is fairly stable on exposure to plasmin. According to the results mentioned above, the mechanism of the plasmin digestion of factor XIII becomes highly complex by the inhibitory effects of the combination with subumit S or plasma protain including fibrinogen.
It may be tentatively explained that the decrease of the factor XIII in afibrinogemia and DIC may be due to the consumption of factor XIII during the process of blood coagulation, and the plasmin digestion of the subunit A in the tissues and in the plasma by the local or secondary fibrinolysis with diminished plasma fibrinogen concentration.

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A new technique for factor XIII determination, spot fluorescent method (K. Aizawa et al., SEIKAGAKU, vol. 46, 568, 1974, abstract) effectively used screening many test samples in normal, in arteriosclerotic patients, and in pregnant women. The results were summarized as followings.
1) Plasma factor XIII concentration in normal persons is gradually increased depending on age from teenage to older, and rapidly increased from late middle age. There is obvious positive relationship between plasma factor XIII and age.
2) The avarage and the standard deviation in young adult was 18.4±3.0
3) There was little relationship between factor XIII and puls wave velocity which was considered to indicate the extent of aortic sclerosis. But factor XIII in the persons who gave myocardial disorders i. e. abnormal ST, T in the ECG were meaningfully high, while the broad distribution from low to high was seen in the patient group of angina on the other hand.
4) Every malignant tumor and liver cirrhosis showed abnormally low level of plasma factor XIII.
5) Fibrinogen, platelet count, and total protein were all once decreased by pregnancy, and became recovered from 1st trimester, 2nd trimefter, or 3rd trimester stage individually.
On the other hand factor XIII was increased by pregnancy and maintained in the high level during all trimester, and rapidly decreased after delivery.

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Plasma factor XIII of babies and children were surveyed to determine normal range in each growth stage and the variation in several diseases were observed.
To investigate the susceptibility of factor XIII by the effect of intravascular accident, thrombosis for instance, the experiment of laurtc acid-induced intravascular disorders with rabbits was carried out. The results were as follows.
1) Concentration of factor XIII in cord blood was extremely low (7.7±2.6).
2) In normal case, the level of factor XIII was the lowest (9.9±2.6) in new born baby, going up to the highest (22.0±5.6) in infancy, then slowly down to 18.0±7.7 in pre-school age, and 13.2+5.3 in school age.
3) Plasma factor XIII was relatively increased in respiratory infections especially in streptococcosis, and was greatly varied from abnormally low to very high value in MCLS and ITP.
4) Lauric acid was intrvenouly injected into rabbits according to Zbinden's experiment. By this effect plasma factor XIII was gradually decreased, while platelet count and fibrinogen level were pronouncedly altered.

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In our present report, the factor XIII of newborn was 55.8, SD±13.5% by clot solubility method, 56.4, SD±19.5% by immunological method and 10.4, SD±2.17μM MCD/10min by fluorescent method. These low values of newborn were compared whith normal adult values of 100% or more, 108.6%, SD±17.8% and 17.55, SD±4.32μM MCD/10min respectively. The factors which caused low in the values of newborn were investigated. Firstly, in the cases of newborn, the restoration of low value of factor XIII by the adittion of cysteine in vitro is only partial and not complete as the cases of liver diseases. Secondary, the results of the experimental studies which trans-AMCHA was administered to the pregnant women before the term and their newborns after the birth, showed as follows. In the cases of the newborns of two days old, both the activity of plasmin and the value of FDP were remarkably decreased, while the factor XIII in administered group was unchanged or rather lower than the control. Our results may indicate that other factors than increased fibrinolysis would contribute to the low values of factor XIII during newborn period.

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This report presents a congenital factor XIII deficiency with platelet dysfunction.
R. S., a 18-yr-old Japanese girl, experienced the first episode of bleeding at age of 4 days, with a sever and prolonged umbilical vein hemorrhage. Since childhood she had repeatedly suffered prolonged bleeding after cut, or dental extraction. She also exhibited slow healing of wounds. She was admitted to hematologic clinic in Oct. 1973 because of hemorrhagic diatheses. The family history revealed no consanguineous marriage. No members of her family exhibited any hemorrhagic manifestations.
No abnormality was found in various laboratory test except for thrombelastogram and clot lysis test. Patient's plasma clot in 5M urea or 1% monochloracetic acid solution dissolved within 60 minutes. Assay of factor XIII level by antiserum inhibition method revealed low level less than 2% in the patient. Factor XIII level of father, mother and brother was 80, 100 and 100%, respectively.
Gel diffusion test disclosed the absence of factor XIII subunit A in patient's platelets.
Platelet aggregation study revealed moderately impaired aggregation of patient's platelet in employing ADP (final conc. 2.5-5×10^-6M), collagen (6μg/ml), adrenaline (5.5×10^-5M), thrombin (0.25U/ml) or bovine fibrinogen (0.1-0.2mg/ml).
Platelet retention test by Hellem II glass beads column showed normal retention.
Amount of patient's platelet factor 3 and 4 after destroying platelets was within normal limits. Platelet factor 3 availability test using Kaolin-Stypven time method disclosed slightly impaired factor 3 availability.
The amount of released platelet factor 4 during aggregation of patient's platelet induced by ADP (final conc. 1×10^-5-10^-4M) or collagen (2-15μg/ml) was always less than 1%.

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In 1944 Robbins reported the existence of factor XIII, and thereafter many investigators have reported the role of factor XIII in coagulation mechanism.
This enzyme has, also, some action on the atherosclerotic changes in arterial walls. In order to clarify the relation between the enzyme activity and the atherosclerotic severity, we measured the activity of factor XIII and transglutaminase in human aortas.
Methods and Materials
Relatively fresh 10 human aortas were selected from forensic autopsy. The atherosclerotic severity (0 to 4 stage) were calculated on macroscopic findings. The intima-media preparation was removed from aortic arch, thoracic and abdominal aortas. Measurement of enzyme activity;
(1) Partigen method: In each plate, 0.2ml of Anti factor XIII subunit serum A (Anti-A) and Anti factor XIII subunit serum S (Anti-S) were separately added to 14ml of 1.5% agarose solution at 52°C to 55°C. 20% homogenate of samples were kept at 4°C in refrigerator for 24h-dialysis. Then after centrifugation of the homogenate at 10000rpm for 15min., 40μl of the supernatant was injected in each hole (diameter 3mm, 12 holes in one plate) and then kept in room temperature for 48h. Factor XIII was shown by the calculation of each diameters.
(2) Transglutaminase activity
We used the modification of Dvilansky et al. Cadaverine-1, 5-¹⁴C-dihydrochloride (0.25μCi) and 1ml of 10% homogenate were incubated at 30°C for 4h. 0.1ml of aliquots were taken into 5ml of 5% TCA at 0, 30, 60, 120, 180 and 240min. Each sample was rinsed with 5% TCA and filtered on Millipore filter.
Results
In Partigen method, we found factor XIII in all stages of atherosclerosis. In early stages factor XIII seemed to be higher, but there were no significant differences.
In transglutaminase activity, we found the relatively higher counts in stage 0 to 3. In stage 4, however, there was lower counts, which means the lower metabolic changes in the highly damaged.
Conclusion
We investigated factor XIII and transglutaminase activity in human aortas. In early stages of atherosclerosis, we found the some degree of enzyme activity. Laki et al. reported the similar results in animals, in which they could not ascertain whether they were dealing with the tissue or the plasma enzyme. We also imagine that the large parts of enzyme in arterial wall may come from blood plasma, but some parts of it come from the tissue. Factor XIII may have both in blood plasma and in arterial walls.

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Factor XIII activities of 36 patients with hepatic disease, i. e. cirrhosis (18 cases), chronic hepatitis (10), acute hepatitis (6 including those in the recovery stage) and obstructive jaundice (2), were estimated by the quantitative test using anti-Factor XIII serum. Also relationship of level between Factor XIII and some such factors as the following was studied in hepatic diseases described above: platelet, fibrinogen, FDP and various substances for liver function tests.
Most cases of cirrhosis and chronic and acute hepatitis revealed low levels of the Factor XIII activity, but 2 of obstructive jaundice showed high activities of the factor.
A positive correlation was roughly present between the Factor XIII activity and platelet count, suggesting some relation of the platelet content to the factor in study or possible effect of the defibrination syndrome, the existence of which in hepatic disease has affirmatively been presumed in these years.
Contrary to expectation, however, correlation was little between the Factor XIII activity and fibrinoeng or FDP levels.
Among liver function tests, positive and negative correlations were seen of the activity in question respectiivly to cholinesterase and ICG, though both slightly. As the two tests are primarily concerned with indication of hepatic cellular disorder, the role of the liver in Factor XIII production may be anticipated from these results, to a certain degree.
Factor XIII activities fluctuated more or less according to cases along with their clinical courses.

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The significance of the Factor XIII for wound-healing is not yet generally accepted. Our tudies of surgical patients indicate possible connection between Factor XIII and wound-healing, because marked postoperative reduction of Factor XIII is supposed to disturb it. Patients with delayed wound-healing or rupture of sutured wound show marked reduction of Factor XIII, but patients with marked reduction of Factor XIII do not always show delayed healing of them.
Our clinical studies revealed that reduction of Factor XIII of patients without surgery is possibly originated, on the one hand, partly by disturbances of liver functions and, on the other hand, partly by appearance of endogenous wound such as gastrointestinal ulcers, resulting proteolytic processes.
Our experimental studies showed that experimental reduction of Factor XIII of rat was achieved with administrations of liver-intoxicating drugs such as d-galactosamine (200mg/kg) or CCl₄ (2ml/kg) and also achieved with exposure of resticted rat to cold environment.
Vitamine K₂, the drug against liver function disturbances and against gastrointestinal ulcerations have showed benefical effects to the reduction of Factor XIII resulting the delayed wound-healing and appearances of gastrointestinal lesions.
A relation between Factor XIII concentration and delayed wound-healing or appearance of endogenous wound was demonstrated in our clinical and experimental studies. The lower the Factor XIII concentration, the more were the delays in wound-healing or number of ulcers as endogenous wound.

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Immobilized asparaginase was prepared by embedding asparaginase (which is effective for remission children with leukemia) into fibrin polymer formed by fibrinogen-fibrin conversion in the presence of thrombin. The immobilized asparaginase film did not dissolve in 6M urea, suggesting that blood coagulation factor XIII participates in the cross-linking between fibrins and between fibrin and asparaginase.

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