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A high performance liquid chromatographic (HPLC) method with electrochemical detection (ECD) was developed for the simultaneous measurement of estrone, estradiol, estriol and estetrol in serum. These hormones were extracted with diethylether, chromatographed on an silica-octadecyl silane (ODS) column with an eluent of phosphate buffer solution-acetonitrile-methanol (volume ratio 152:85:40), and detected by ECD at +1.0V vs. Ag/AgCl. In comparisons between the values measured by this method and radioimmunoassay, significant correlations were noted for estrone (r = 0.759, p less than 0.01), estradiol (r = 0.816, p less than 0.001) and estriol (r = 0.830, p less than 0.001). In clinical applications of this method, differences between cases of the normal and the anencephalic pregnancy in the thirty-eighth week of gestation were distinct not only in the individual estrogen, but also in the profile analysis of estrogens. With this method, all 4 serum estrogens above approximately 500 pg/ml could be measured within 2 h, and the method seemed to be clinically applicable.

The results obtained in this investigation may be summarized as follows : 1. The CO2-output of the male muscle and other tissues is greater than that of the female. 2. The female muscle contains larger amount of water than the female muscle. 3. The muscle immersed 1/2 Ringer solution (or 1/2.5 R.) gave out smaller amount of CO2 per minute than the muscle in 2-Ringer's solution (or 2.5 R.). In spite of the difference in the water content of tissue between different sex, the salt content of the tissue liquid seems to be the same. In other words, larger the water content means larger content of tissue liquid in the tissue. Artificial introduction of water in the tissue or reduction of water content by immersing the tissue in 1/2 or 2-Ringer's solution is quite different from the natural condition occurring between different sex. However both of these conditions influence the gaseous metabolism in the same manner. On an assumption that the gas diffusion in liquid is proportional to the solubility of that gas, the above mentioned difference of CO2-output should be just reversed. Therefore it is not possible to interpret how the water content influences the gaseous metabolism. It may only be stated that the muscle which has a small amount of water to an extent which does not abolish excitability, gives out much CO2 and vice versa.

1. The properdin levels in sera from mice bearing Ehrlich ascitic carcinoma and from rabbits with Brown-Pearce carcinoma decrease inversely with the increase of the ascites or the tumors. In the incipient period of tumor transplantation, the level rather rises. When the tumor is proliferating or large, the level keeps falling or is low. On the contrary, when the tumor is regressing or disappears, the level elevates or reverts to that before transplantation. Strong A and R III mice with spontaneous mammary cancer have markedly low serum properdin levels as compared with those of healthy mice. 2. The properdin levels are less than 2 units per milliliter of the serum in 44.4 per cent of patients with gastric cancer, in 18.2 per cent of ones with non-malignant tumor and in 18.2 per cent of ones with gastric or duodenal ulcer. The abnormal low level has been found in 33.3 per cent of patients without recurrence, who had undergone extended radical gastrectomy combined with radical lymphadenectomy for gastric cancer. 3. Some correlation can be seen between the serum properdin levels and the degree of progress of gastric cancer. 4. The cancer patients with low total serum protein have lower serum properdin levels than those having nomal protein. 5. As for influence of surgical operation on the serum properdin levels, there is observed a tendency that a minor operation causes the levels to increase and a major operation causes the levels to fall. 6. It has been inferred that the properdin system could be one of the host natural resistance against cancer.

Neomyographic examinations were made by the authors on 28 patients. The extent of reconvalescence was measured on the basis of changes in the values recorded by the myograms taken before and after the treatment.

The human tumor H. Ep. #3 maintained in rats could be transferred for 1-8 generations in treated guinea pigs. H. Ep. #3 grew in the subcutaneous and intramuscular sites in each host at the same time. The treatment with the combination of X-ray 250 r. and 80 mg/kg of cortisone turned out to be the optimal conditioning studied. The number of tumor takes averaged 95.7-100 per cent in the subcutaneous site in guinea pigs treated with optimal conditioning, but in the intramuscular site, the number of tumor takes was 65.2-93.8 per cent. Host mortality varied from 4.2-37.5 per cent in the hosts treated with optimal conditioning. The subcutaneous tumor weights in hosts treated with optimal conditioning averaged 3.3 gm, and their intramuscular tumor weights averaged 5.6-6.2 gm. Tumor weights in hosts treated with only cortisone averaged 1-2 gm in both subcutaneous and intramuscular sites. Histological findings. for the original tumors were found to be the same as that for the successful transplanted tumors in the guinea pigs. The malignancy of the tumor was evaluated by the criteria of anaplasia, invasion, rapidity of growth, and ease of maintenance of transplants. There was no metastasis found in any organs.

Die Autoren geben bekannt, daβ das SZIRMAI'sche Myotonometer nicht nur in der Diagnostik der Stomatologie auch von Bedeutung ist, sondern man ist imstande mit Hilfe des Apparates nicht nur auf dentalem Gebiet, sondern auch bei anderen verschiedentlichen Krankheiten die Einwirkung der Medikamente auf die Kaumuskel bzw. auch auf den Musculus Masseter aus zu werten. Somit konnen wir bei der zahnarztlichen Therapie die objektivste Behandlungsmethode erwahlen.

With the purpose to study sideroblasts as a means of diagnosing blood diseases and to pursue the metabolism of non-hemin iron in erythroblasts we investigated sideroblasts (erythroblasts containing iron granules stainable by Prussian blue) under various erythropoietic conditions in the human and rabbits, and obtained the following results: 1. In blood diseases the proportion of sideroblasts in the case of low erythropoietic condition is higher and in the case with accelerated erythropoietic condition and of iron deficiency it tends to be lower than that in normal persons. Further, obtaining sideroblastogram and sideroblast ratio (S. r.) from the classification of Types I, Ⅱ, and Ⅲ according to the iron granule content, it has been proven that abnormal conditions can be clearly distinguished from the normal, indicating that sideroblasts are closely associated with erythropoietic function. This is proven to be a far superior method for the diagnosis as well as for the prognosis of blood diseases than the determination of serum iron. 2. In experimental anemic rabbits the relationship of sideroblasts to the condition of erythropoietic function is still more clearly recognized, and it has been found that variations in the sideroblast count is dependent upon the condition of the equilibrium between the iron supply from serum iron and the iron utilization controlled by the erythropoietic function. 3. In addition, in the iron-treated rabbits under various erythropoietic conditions we have been able to confirm that there are a certain mechanism and a limitation to the iron intake by erythroblasts, and that erythroblasts take essentially three steps of metabolic processes, namely, intake, retention, and utilization of iron almost simultaneously, in the latter half of the maturation stage.

Die Autoren haben mit Hilfe des Szirmaischen Myographen auf Grund der Messung vor und nach der Behandlung den Reaktions-Index aus den Myogrammen erhalten. Mit dieser Methode laβt sich in einer sehr kurzen Zeit bei verschiedenen Krankheiten die gunstigste Therapie auswahlen und deren Wirksamkeit in Prozenten auswerten.

Hepatocyte growth factor (HGF) and c-met proto-oncogene product (c-Met) have varied biological functions in different tissues and have been implicated in mitogenic, motogenic and morphogenic responses in both organ regeneration and carcinogenesis. Some studies have suggested that the overexpression of c-Met and epidermal growth factor receptor (EGFR) are associated with growth advantage, while transforming growth factor-beta receptor II (TGF beta R II) is associated with growth disadvantage of human prostatic adenocarcinoma. However, it is unclear if the expression of c-Met correlates with the expression of EGFR and TGF beta R II, and with the proliferative status of human prostatic adenocarcinoma. Using immunohistochemical staining with anti-c-Met (C-12), anti-EGFR (NCL-EGFR) and anti-TGF beta R II (L-21) antibodies, we determined the frequency of expression of c-MET, EGFR, and TGF beta R II respectively in a series of 134 radical prostatectomy specimens. We evaluated the relationship between the expression of these receptors and clinicopathological characteristics. Overall, c-Met immunostaining was detected in 54 of 134 (40.3%) cases, EGFR in 45 (33.6%) and TGF beta R II in 64 (48.4%). The overexpression of c-Met was significantly more common in poorly differentiated (P < 0.0001) and in the diffusely infiltrated specimens (P < 0.0005). In contrast, TGF beta R II was significantly overexpressed in the well differentiated specimens (P < 0.0001) and associated negatively with c-Met (P < 0.0001). Overall, these data suggest that c-Met/HGF receptor and TGF beta R II overexpression may be involved in the differentiation of human prostatic adenocarcinoma, c-Met with de-differentiation and TGF beta R II with differentiation.

For the purpose to reveal the role of R.E.S. for hemopoiesis and antibody formation, the R.E.S. of rabbits were severely blocked by the repeated intravenous injection of a vast amount of India ink, reaching 200 to 250 cc in total and the development of anemia and antibody formation by challenging egg albumin were observed while referring to the histologic changes in bone marrow, spleen and lymph nodes. The results were as follows: 1. The repeated intravenous injection of a vast amount of carbon particles induced a severe anemia. The anemia was always normo- or hyperchromic, showing not any disturbance in iron metabolism or hemoglobin formation. The data suggested that anemia is due to the arrest of reproduction of erythroblast or differentiation of the stem cells to erythroblasts, but not due to inhibition of the iron metabolism. 2. R.E.S. had no relation to the proliferation or the differentiation of granulocytes. 3. The functions of R.E.S. related to erythropoiesis and lymphopoiesis are affected by blocking independently of its phagocytic potency. In spite of a severe anemia, the phagocytic potency of R.E.S. could never be lowered and liver and spleen grew much larger in size and weight, showing that the phagocytic ability of R.E.S. is extremely resistant against such a blocking. 4. The serum antibody titer proved to be at the normal level, and no change of the antibody production in spite of heavy blocking of R.E.S. with India ink.

In vitro cell transformation of human embryo cells could be induced by the DNA purified from SV 40. The result shows clearly that cell transforms a part of viral DNA into the genome. In addition, for the purpose of clarifying th~ biological differences between the normal and transformants the alteration of cell membraneous structures of transformants (hamster and mouse fibroblasts) were observed from mechanism of phagocytosis. The iron colloid particles are taken up by normal diploid fibroblasts but not by the human and hamster transformants. This fact suggests a differ~nce in the molecular arrangement of the cell membranes between the normal and transformants. In the presence of histones, however, the transformants phagocytize the colloid particles very actively. The results show that cell membranes of transformants are altered in the molecular structure r~sponsible for the surface charge. In addition, there is no remarkable quantative differences of sialic acids on the cell surfaces of non-malignant and malignant transformants so that phagocytic activity might be correlated to the alteration of molecular composition of cell membrane itself rather than of cell surfaces, i. e, sialic acids.

Superoxide anion (O2-) production by neutrophils from 14 untreated patients with acute nonlymphocytic leukemia (ANLL) was significantly less than that of healthy controls (4.93 +/- 1.99 vx 6.20 +/- 1.53 nmol/min/10(6) neutrophils, p less than 0.05). In 10 patients with myelodysplastic syndrome (MDS), however, it was not significantly different from the control level although 6 of the 10 patients had low levels, when individual patients were compared with the lower limit of the control range. An inverse correlation between the O2- production of neutrophils and the percentage of leukemic cells in the marrow existed in ANLL (r = -0.55, p less than 0.01), but not in MDS. Three of 4 MDS patients who died of pneumonia prior to leukemic conversion showed a low level of O2- production. The impaired O2- production by neutrophils from some MDS patients, probably due to the faulty differentiation from leukemic clones, may be one of the causes of enhanced susceptibility to infection.

The stability of recombinant human superoxide dismutase (r-hSOD) in buffer solutions was studied in solutions at various pH and temperatures. Additionally, we studied the effects of incubation with proteases, serum and two types of hypothermic perfusates. R-hSOD was stable in the pH range of 6-11 and at temperatures up to 80 degrees C for 30 min. R-hSOD activity was not affected by incubation with trypsin, aminopeptidase M or serum for 2 h. R-hSOD activity determined at various temperatures (4-37 degrees C) did not vary remarkably. R-hSOD in hypothermic perfusates was stable at 4-37 degrees C for 24 h.

The vascular effects of an aqueous extract prepared from the leaves of Eucommia ulmoides Oliv. (ELE), a medicinal herb commonly used in antihypertensive herbal prescriptions in China, were investigated in rat mesenteric resistance arteries. The mesenteric vascular bed was perfused with Krebs solution and the perfusion pressure was measured with a pressure transducer. In preparations with an intact endothelium and precontracted with 7μM methoxamine, perfusion of ELE (10^－7－10^－2mg/ml for 15min) caused a concentration-dependent vasodilatation, which was abolished by chemical removal of the endothelium. The ELE-induced vasodilatation was inhibited by neither indomethacin (INDO, a cyclooxygenase inhibitor) nor N^G-nitro-L-arginine-methyl ester (L-NAME, a nitric oxide inhibitor). The ELE-induced vasodilatation was significantly inhibited by tetraethylammonium (TEA, a K⁺ channel blocker) and 18α-glycyrrhetinic acid (18α-GA, a gap-junction inhibitor), and abolished by high K⁺ -containing Krebsʼ solution. Atropine (a muscarinic acetylcholine receptor antagonist) significantly inhibited the vasodilatation induced by ELE at high concentrations. These results suggest that the ELE-induced vasodilatation is endothelium-dependent but nitric oxide (NO)- and prostaglandin I₂ (PGI₂)-independent, and is mainly mediated by the endothelium-derived hyperpolarizing factor (EDHF) in the mesenteric resistance arteries. Furthermore, the ELE-induced EDHF-mediated response involves the activation of K⁺-channels and gap junctions.

Our study aimed to investigate the potential radioprotective effects of N-acetylcysteine (NAC) by comparing its biochemical effects with those of WR-2721, as a representative of clinically used radioprotectors, in preventing oxidative damage caused by gamma irradiation (single dose, 6Gy) in normal rat tissue. The rats (n=40) were divided randomly and equally into 4 groups:Control (C), Radiation (R), R+NAC (received irradiation and 1,000mg/kg NAC) and R＋WR-2721 (received irradiation and 200mg/kg WR-2721) rats. Liver tissues and blood samples were harvested and utilized for reduced glutathione (GSH), malondialdehyde (MDA) and myeloperoxidase (MPO) detection. Serum and tissue GSH levels of R rats decreased compared to those of other groups (p<0.01). Tissue MDA levels of R+NAC and R+WR-2721 rats decreased compared to R rats (p<0.01;p<0.05, respectively). Tissue MPO activities of R+NAC and R+WR-2721 rats were higher than those of R rats (p<0.001). Serum MPO levels of R+WR-2721 rats were lower than those of C rats and R rats (p<0.01, p<0.001, respectively). In conclusion, the study suggests that the radioprotective effect against radiation-induced oxidative damage of NAC may be similar to that of WR-2721.

Glucocorticoid effect on thyrotropin-releasing hormone (TRH)-induced prolactin (PRL) release was studied in female patients with collagen or autoimmune diseases. Long-term, high dose glucocorticoid therapy tended to inhibit the response of plasma PRL to TRH. A negative correlation (r=-0.40) was found between the logarithm of total dose of glucocorticoids received and the magnitude of plasma PRL response to TRH (p less than .05).

The plasma fatty acid composition of cirrhotic patients and their dietary intake of fatty acids were determined. Significantly lower plasma arachidonic, docosahexaenoic, dihomo-gamma-linolenic and eicosapentaenoic acid levels were observed in cirrhotic patients than in healthy controls. A remarkably low dietary intake of polyunsaturated fatty acids supplied from fish, vegetable oil and pulses was shown in cirrhotic patients. Positive correlations were observed between plasma arachidonic acid concentrations and clearance rate of indocyanine green (KICG) (r = 0.826, p less than 0.05) and between dihomo-gamma-linolenic acid levels and cholinesterase activities (r = 0.841, p less than 0.05). From these results, we conclude that a supply of polyunsaturated fatty acids is necessary for the nutritional treatment of patients with liver cirrhosis.

The microfilaria of Brugia pahangi obtained from an experimentally infected dog were observed with the electron microscope. The sheath was composed of small granules and was covered with electron-dense particles on the outer surface and with small granules on the inner surface. The cuticle was composed of an outermost layer, a trilaminate membrane and an inner layer. The hypodermis was composed of four components as in the adult stage (two small ones on the lateral sides, two large ones on the dorsal and ventral sides). The muscle cells comprised a single layer under the hypodermis on the dorsal and ventral sides. On each side, two muscle cells usually appeared in a transverse section. The thick myofilament was surrounded with 8 to 12 thin myofilaments. Dense bodies were present around the cephalic space. In the cells of the nuclei column, the cytoplasm was very narrow, and the electron-dense nucleus close to each other. The cuticular central canal was connected to the buccal cavity and to the inner body. A sponge-like structure was seen at the junctional part of the canal and the inner body. The inner body showed a homogeneous granular appearance. Eight cephalic papillae were observed at the head tip. Two amphids, each having more than eight cilium-like structures, were connected with the nerve elements and open in the head part. Two phasmids, each having one ciliumlike structure, opened in the caudal part. Two types of neurosecretory granules were observed in the nerve ring and the dorsal and ventral longitudinal nerves were clear except in the anterior and the posterior part of the worm. The excretory and the anal vesicles had contacts with thin and thick cytoplasmic processes respectively, and these vesicles opened to the exterior. The nuclei of the G cell and R cells showed similar electron-density. Lamellate structures were present in the muscle and the hypodermis.

A study was conducted to examine the relationship of testicular atrophy to bone metabolism in male leprosy patients. The study consisted of 31 leprosy patients (mean age: 62.0 years) and 31 healthy control men (mean age: 60.0 years). Measurements were made of their serum levels of free testosterone (FT), estradiol (E₂), luteinizing hormone (LH) and 25-hydroxyvitamin D (25 OHD). Bone mineral density (BMD) was measured at radial sites and the lumbar vertebral bodies (L2-L4) by dual-energy X-ray absorptiometry using a Hologic QDR-2000 densitometer. FT and E₂ levels were significantly lower and LH levels higher in leprosy patients than in controls. This represents a primary hypogonadal pattern. A value of 7.20pg/ml of FT (= Mean -1 SD of control) was used as a cut off value, and the subjects were subdivided into a hypogonadal group (HG) and a non hypogonadal group (non-HG). When the subjects were compared for differences in age, age at onset of disease, duration of disease, body mass index and BMD, only the duration of disease and BMD were significantly different between the two groups. Furthermore, BMD of the forearm significantly correlated with FT levels (r = 0.689, P < 0.0001). Low BMD may be due to orchitis and testicular atrophy.

Even after successful operations, ugly postoperative skin scars are often distressing to patients and their parents. To judge the success of surgical methods and postoperative treatment, postoperative scars should be evaluated using a quantitative system. Height and width are easily measured, but scar redness is not. We have developed a simple and effective method for evaluating scar redness. According to the color definitions employed in computer graphics, each color can be expressed as RGB (red, green or blue) coordinates (r, g, b): 0 ≦ r, g, b ≦ 10. The degree of scar redness is defined by the following formula: redness score (RS) = (r₁ - r₀)² + (g₁ - g₀)² + (b₁ - b₀)². Here, (R₁, g₁, b₁) = coordinates of the scar color and (r₀, g₀, b₀) = coordinates of the surrounding skin color. RS was evaluated in 59 children (35 males, 24 females; ages 1 month to 12 years old) who had scar redness after congenital cardiac surgery. For each patient, scar color and surrounding skin color was identified on the color sample table. Scar redness was also evaluated by the conventional grading method: 1 = mild, 2 = moderate and 3 = severe. The RS of the colored scars ranged from 4 to 100 (38 ± 27). By the conventional grading method, 44 scars were grade 1, 15 grade 2 and none grade 3. RS was significantly higher among grade 2 than grade 1 patients, 52 ± 25 and 33 ± 27, respectively (P < 0.05). Given its subjectivity, the conventional grading method yields variable data; surrounding skin color, moreover, is not considered. Our new evaluation method using RS effectively and accurately defines scar and skin colors, and allows quantitative studies of these factors.