A freeze-fracture study of hormone-induced branching in the fungus Achlya. The induction of the male sexual organ primordia (antheridial hyphae) by the steriod hormone antheridiol in the water mold Achlya ambisexualis Raper requires the production and secretion of the enzyme cellulase. It is postulated that a localized secretion of cellulase produces a limited area of wall hydrolysis that is blown out into a lateral bleb by turgor pressure. Freeze-etch preparations show membrane profiles similar to those seen in other systems where exocytosis is occurring. Such a mechanism would provide the required localized secretion of cellulase. Water stress, imposed by polyethylene glycol, prevents the formation of antheridial hyphae, the secretion of cellulase and the expected membrane profiles. After a period of recovery from water stress antheridial hyphae are formed, cellulase secretion occurs and the expected membrane profiles are restored.


The screening test of various chemotherapeutic drugs in primary malignant melanoma cells and human malignant melanoma cell line (TM-1). The sensitivities to various chemotherapeutic drugs of the primary malignant melanoma cell of the esophagus and the malignant melanoma cells established from human skin were studied by determining the incoporation of 3H-thymidine into tumor cells using the tissue culture method. The incorporation of 3H-thymidine was depressed by a low concentration of Actinomycin D in malignant melanoma cells of the esophagus and by a low concentration of Adriamycin or Actinomycin D in the established malignant melanoma cells. When Actinomycin D was clinically used according to the experimental results, the subcutaneously metastasized tumor was remarkably reduced.


On a new continuous cell line (TM-1) derived from malignant melanoma. A continuous cell line of malignant melanoma has been established. The tumor specimen was obtained from a left axillary lymph node of a malignant melanoma originating in the abdominal skin of a 67-year-old woman. The cell line has been designated TM-1. Explant specimens showed positive dopa staining, but after 2 years and 4 months, or 60 generations of cell passage, the dopa staining is now weakly positive. Treatment with DBcAMP that causes marked morphological changes induces melanin production and lowers the proliferative ability of the cells. These changes were reversible. Transplantation of the cells to nude mice resulted in the formation of large tumors consisting of epithelioid cells and spindle cells. Morphologically, the majority of the cell population in vitro was polygonal epithelioid cells, but the cell passage produced a large number of spindle cells. The shape of the TM-1 cells was examined by scanning electron microscopy. Depending upon the culture conditions, either of these cell types could be predominated, indicating that both cells are of the same origin.


Immunosuppression induced by "toxohormone" from mouse tumor cells in culture. Tumor culture toxohormone (TCT) obtained from cultures of MBQA mouse tumor cells, a line derived from a methylcholanthrene-induced fibrosarcoma (CBA/J origin), suppressed the mitogenic responsiveness of mouse spleen cells (PHA, LPS) as well as the antibody formation to SRBC in vitro. The immunosuppressive activity of toxohormone was readily inactivated by heating at 100 degrees C or treatment with trypsin, but not by DNase and RNase treatment.


Anti-nucleoprotein antibody response in influenza A infection. The nucleoprotein (NP) antigen isolated from influenza A virus by solubilization with Triton X-100 (TX-100) and further electrophoresis with SDS-cellulose acetate membrane gave a single band on SDS-polyacrylamide gel electrophoresis. Rabbit anti-serum hyperimmunized with the NP reacted only against the NP antigen. Moreover, a well-defined single precipitin line was shown between the NP and human sera. These results suggested that the NP was possible to detect anti-NP antibody in human serum. Immuno double diffusion (IDD) and single radial immunodiffusion (SRD) tests using the NP were established to detect the anti-NP antibody in human sera. During an epidemic caused by antigenic drift strain, anti-NP antibody was detected by the IDD test in the cases which did not show any significant rise in HAI titer. During a mixed epidemic caused by the different strain of HA antigen, the infection ratio in mass population was revealed more convenient and sensitive by SRD than HAI. The anti-NP antibody was detected by IDD for long periods of one year or more after infection. These results suggest that the detection of anti-NP antibody is applicable to serologic studies, particularly serologic diagnosis and serologic surveys of influenza infection in mass population.


Effect of adenosine deaminase replacement therapy on a child of adenosine deaminase deficiency with severe combined immunodeficiency disease. Enzyme replacement therapy was performed for a 1-year and 5-month old boy with adenosine deaminase deficiency disease, the first case in Japan. Irradiated fresh red blood cells were administered without any clinical improvement, but there was an increase in the peripheral lymphocytes from 300/mm3 to 1849/mm3, of which 88% had T cell marker. B lymphocytes did not bear any classes of surface immunoglobulins. The proliferative responses of these lymphocytes to phytohemagglutinin, concanavalin A, pokeweed mitogen and allogeneic cells were examined. More than two-fold increase in response to these mitogens was observed in lymphocytes after treatment as compared with responsiveness before treatment, but these responses still remained to a much lesser degree than that of lymphocytes from controls.


Antimetastatic effect of triton WR 1339, a nonionic detergent, on rat ascites tumors. The antimetastatic effect of Triton WR 1339 (TWR), a nonionic detergent, was evaluated in the rats bearing ascites tumor (Yoshida sarcoma or AH 66F). TWR did not inhibit the growth of primary tumor (ascites production). However, a marked inhibition of metastasis was observed in the lung and liver of rats treated with TWR. The results obtained suggest that TWR exerts antimetastatic effect before the entry of tumor cells into vascular channels. With other tests, it was found that TWR inhibits the release of tumor cells from tumor graft.


The correlation between hypertension in past history and the incidence of cerebral aneurysms. The incidence of hypertension in past history was investigated in 811 cases of cerebral aneurysm. These cases were compiled from 1,000 cases of saccular aneurysm in which direct surgical operations for aneurysm were performed at our clinic during the period from June 1961 to September 1975. Of the 811 cases, 365 (45%) had hypertension in their past history; 185 (42.7%) out of 433 males and 180 (47.6%) out of 378 females. In the 5th decade of age, the incidence was significantly higher in the females than in the males, but no difference by sex was noted in other age groups. In the males from the 3rd to the 7th decade, the number of hypertensives increased significantly with advancing age, whereas in the females a significant difference was observed only between the 4th and 7th decades, the latter including more hypertensives. The incidence of hypertensives in the aneurysm cases was compared with that in the Japanese population reported by Sasaki. This comparison revealed that in both sexes between the 4th and 6th decades, the incidence was significantly higher in the former, whereas no significant difference was noted between the two in the 7th decade. As to the location of aneurysms, only the multiple aneurysms group had a significantly greater number of hypertensives than single aneurysm group. These results agree with previous reports that cerebral aneurysm may occur more frequently in the hypertensives than in the normotensives.


Filtrability and flow characteristics of leukemic and non-leukemic tumor cell suspension through polycarbonate filters in relation to hematogenous spread of cancer. Tumor cell suspension was filtered through Nuclepore filters of various pore diameters (5.4, 7.9 and 9.3 micron) with positive pressure from 5 to 60 cmH2O at 37 degrees C. The mean diameters of tumor cells of 6 strains ranged from 10.6 to 13.6 micron. Cell suspension of each tumor strain was filtered with characteristically different time. No significant difference was observed among tumor strains in the percentage of cells filtered. The cell viability was almost unchanged by filtration. The filtration time was considered to indicate the passing ability of tumor cells through capillary pores. Leukemic cells such as DBLA 1, DBLA 6 and L 1210 were relatively small in diameter and possessed a high passing ability compared with other non-leukemic tumor cells such as Yoshida sacroma, AH 109A and AH 100B. The relationship between pressure and flow rate of the cell-free solution was linear, while the pressure-flow rate curves of the tumor cell suspension were convexed to the pressure-axis at low pressure and became linear over the pressure of the yield point. Rheologically, the yield point and the reciprocal of the slope indicate structural viscosity and apparent viscosity of the cell suspension, respectively, they are considered to reflect the rheological properties of tumor cells. Comparing these parameters of the curves in filters of different pore diameters, the viscosity of leukemic cells appeared to be the lowest and the structural viscosity of AH 100B cells was the highest among the tumor strains examined. The distribution and frequency of metastases following intravenous transplantation of these tumor cells suggested that the passing ability of tumor cells plays an important role in organ preference of hematogenous metastasis and leukemic state in leukemia.


Experimental and clinical studies on toxicity of xenogeneic tumor-specific immune ribonucleic acid. To learn the toxicity of xenogeneic tumor-specific immune ribonucleic acid (I-RNA), experimental and clinical studies were carried out. Experimentally, doses of 30 mg/kg, 15 mg/kg, 7.5 mg/kg or 3.75 mg/kg of xenogeneic I-RNA extracted from lymphoid tissues of rabbits sensitized with 105,000 Xg sediments of human gastric carcinoma tissue were injected intraperitoneally into Wistar rats once a day for 30 days. During the period of the study, changes of physiological and biochemical values were examined. In addition, pathological study was made for each organ after the I-RNA administration. There was no death during the study. In the groups of high dose administration, there were poor increase in the body weight, elevation of GOT, GPT and LDH, findings of vacuolar degeneration of hepatic cells, and increase of mesangial matrix and polynuclear glomerulus. Throughout the experiment, the groups given 7.5 mg/kg and 3.75 mg/kg of I-RNA showed no significant difference from control groups. Clinically, 31 cases treated with passive immunotherapy with allogeneic lymphocytes, preincubated with xenogeneic I-RNA, were examined retrospectively for the difference in hematological and biochemical data before and after the therapy. One case showed a transient mild febrile reaction after the therapy. There was no significant difference in hematological and biochemical data. Pathological findings in 3 autopsies after the treatment were reported.


Significance of Erwinia in the vagina as causative agents of urinary tract infections. The isolation of Erwiniae from the vagina became positive just before the occurrence of erwiniuria. It is proposed that the appearance of Erwiniae in the vagina is an important indicator in the prediction of urinary tract infection by Erwiniae.


Trophic effect of tetragastrin on the stomach, duodenum and pancreas in rats. Subcutaneous injections of a large dose of tetragastrin (2 mg/day) into a rat for 4 weeks caused hypertrophy of parietal cells of the stomach, intestinal glandular cells of the duodenum, and pancreatic acinar cells. Ths histometrical analysis revealed that these trophic effects of tetragastrin were produced in varying degrees in different sites.


Morphological alteration of aortic wall and mitotic cells after complete endothelial loss induced by repeated balloon denudation of swine aorta. The aortic endothelium in weanling swine was rubbed ten times with an inflated balloon catheter in a repeated balloon denudation. This procedure produced more drastic, extensive and uniform changes in the aortic wall than the commonly used balloon denudation. Sequential alternations of regenerating endothelium, intimal thickening and medial reaction, and the incidence of mitotic cells were studied by scanning and transmission electron microscopy. Complete endothelial loss was confirmed from the descending thoracic aorta to the right femoral artery within 24 hr. By the third day, regenerated endothelium began to cover over the endothelial defect area from the uninjured areas such as the aortic arch and the orifices of branching arteries. Thrombus formation and fibromuscular intimal thickening were observed in the endothelial defect areas by the fifth and seventh days. Three types of mitotic cells, such as endothelial, intimal and medial cells were noted in the aortic wall. Mitotic endothelial and mitotic medial cells were most frequent at Day 3; the latter were closely associated with dead medial cells. Mitotic intimal cells initially appeared at Day 3 and were most frequent at Day 7. Mitotic intimal and mitotic medial cells were frequently present in the aortic wall subjacent to the endothelial defect areas containing interstitial blood components. The large numbers of mitotic aortic cells indicated that endothelial cells give rise to new endothelial cells, intimal cells to new intimal cells, and medial cells to new medial cells.


Residual blood volumes in organs of pregnant mice and fetuses. The supernatant solutions of homogenates of organs of pregnant mice and fetuses on day 14 and 17 of gestation were measured for hemoglobin (Hb) content after a maximal blood extraction at sacrifice. Residual blood volumes of the organs and fetuses were calculated from the Hb contents and blood Hb concentrations. The greatest value was found for the spleen and the lowest for the brain. The residual blood volumes of the uterus, placenta and fetuses on day 17 were greater than on day 14 of gestation.


Response of extrapancreatic glucagon to arginine in dogs. In order to investigate secretion of extrapancreatic glucagon in dogs, plasma glucagon and total immunoreactive glucagon (IRG) in response to arginine were determined with antisera specific and non-specific for pancreatic glucagon, respectively. Only a tiny rise of plasma glucagon was observed in the portal vein following arginine infusion performed immediately after total pancreatectomy in a group of 5 normal dogs. In contrast, total IRG in the portal vein increased significantly after arginine infusion even after pancreatectomy. In alloxan diabetic dogs, arginine infusion after total pancreatectomy caused a rise in plasma glucagon in the portal vein, although not significantly. The response of total IRG to arginine in the portal vein was exaggerated in alloxan diabetic dogs. In a group of one-week post-pancreatectomized animals, plasma glucagon and total IRG increased significantly in response to arginine infusion. Furthermore, in these dogs, response of plasma glucagon and plasma total IRG to arginine infusion was abolished after gastrectomy. From the present results it is concluded that secretion of extrapancreatic glucagon increased in response to arginine infusion in the diabetic state, both alloxan diabetic dogs and one-week post-pancreatectomized dogs. In addition, a rise in extrapancreatic glucagon following arginine infusion in the chronically pancreatectomized dogs almost certainly derived from the stomach, as the rise was abolished by gastrectomy.


Cellular changes in severe dysplasia of the uterine cervix progressing to malignancy. Of a total of 1321 cases of severe dysplasia of the uterine cervix, 237 lesions (18%) were found by punch biopsy and cytological examination to have progressed to carcinoma in situ or microinvasive carcinoma later in the course of follow-up. The changes in exfoliated cells during the progression to malignancy were examined in 95 of the 237 cases. Results obtained were as follows: (1) The ratio of parabasal (immature) cells to the whole dysplastic cells gradually increased in each specimen. (2) Immature dysplastic cells showing increased nuclear membrane tension and irregular staining of the nuclear membrane gradually increased. (3) The number of immature dysplastic cells with finely or coarsely granular chromatin patterns gradually increased.


Improved oral excretory cholecystocholangiography. A new improved technique for the visualization of intrahepatic bile ducts with oral contrast agent has been developed. The right lateral decubitus position was employed to avoid drainage of contrast medium from the common bile duct into the duodenum. 0.2-0.3 microgram/kg of Caerulein was injected intramuscularly (350 cases) or 0.02 microgram/kg of Kinevac was injected intravenously (15 cases) to induce rapid contraction of the gallbladder. The extruded contrast medium, which would normally flow into the duodenum, flowed backward into the intrahepatic bile ducts due to gravity. With this technique, improved visualization of intrahepatic bile ducts was achieved with use of oral contrast medium; it also obviated the problem of colonic gas overlapping the gallbladder and gallstones in cholecystography.


Serum Zn content in tumor-bearing rats treated with anticancer drugs. The serum Zn content in rats bearing Yoshida-Sarcoma, AH 66F, AH 13 or AH 109A was found to be low as compared with that in normal controls. In the rats which had received intraperitoneal inoculum of Yoshida-Sarcoma, the effects of anticancer drugs were closely related to the changes in serum Zn content. Namely, serum Zn content increased and approached the normal range almost parallel with the prolongation of survival time by anticancer drugs. As the causes responsible for the reduction in quantity of serum Zn in the tumor-bearing rats, the reduction of serum albumin content and the accumulation of Zn in the tumor tissue and organs such as the liver and kidneys were suggested.


Abnormal urinary excretion of sialoglycoconjugates in patients with mucolipidosis. Low molecular sialoglycoconjugates were isolated by the charcoal adsorption method from the urines of patients with mucolipidosis (one patient with mucolipidosis type I variant and three patients with mucolipidosis type II). The sialoglycoconjugates were fractionated into two major fractions (SG-1 and SG-2) by Sephadex G-25 gel filtration, and a strikingly increased excretion of the SG-1 fraction was observed in patients with these diseases. The SG-1 fraction from each type of mucolipidosis was fractionated into four subfractions by Sephadex G-50 gel filtration, and the fraction with the lowest molecular weight (SG-1-III) was found to contribute to the increase in the levels of SG-1. The SG-1-III fraction was further fractionated into 7 to 8 subfractions by ion-exchange chromatography. The comparison of the amounts and the chemical compositions of the subfractions suggested that the increase in SG-1 was dependent upon the increase in excretion of mannose-containing sialooligosaccharides with the structures closely related to the carbohydrate units of glycoproteins with an N-glycosidic linkage.


Gonadotropin in follicular phase in women with luteal phase defect. Serum FSH and LH in women with luteal phase defect were measured. Their serum FSH and LH were lower than those in normal women and higher than those in women with amenorrhea. Administration of clomiphene citrate to the women with luteal phase defect were effective in 2/3 of the cases. These results are consistent with the hypothesis that a relative deficiency of FSH during the follicular phase results in diminished follicular development and subsequent inadequate corpus luteum maintenance.


Morphometrical analysis of the peripheral nerve lesions in experimental diabetes rats. Morphometrical analysis of the peripheral nerve lesions in experimental diabetes rats was undertaken to elucidate their relationship to glucose intolerance and nerve conduction velocity. Motor nerve conduction velocity of the rat's tail and nerve fiber density in the cross sectional view of the peroneal nerves significantly reduced in the diabetes rats without insulin treatment. With the teased nerve fiber methods, degenerated nerve fibers frequently appeared and shortening of internodal length was estimated in the diabetes rats by quantification. The slowing of nerve conduction, blood glucose levels on the oral glucose tolerance test and the incidence of degenerated nerve fibers were well correlated with one another. Daily insulin treatment caused a significant decrease in degenerated nerve fibers in the diabetes rats. The reduction of nerve fiber density was also related to glucose intolerance. However, there was neither the relationship between nerve fiber density and incidence of degenerated nerve fibers, nor the relationship between nerve fiber density and nerve conduction velocity.


On the receptors of human red cells reacting with Phaseolus coccineus L. lectin. In order to elucidate receptors of proteolytic enzyme-treated red cells which react with Phaseolus coccineus L. lectin, the receptors prepared by affinity chromatography were serologically investigated. P. coccineus lectin had high agglutinin activity for bromelin-, papain- and pronase-treated red cells but that for the cells treated with ficin and trypsin was relatively low. Analyses of chemical composition revealed that sialic acid of the receptors from normal red cells was considerably much as compared with that from the treated cells. On the contrary, the enzyme treatment did not affect particularly carbohydrate composition of the receptors. Disc electrophoresis showed that the patterns of receptors from red cells treated with bromelin or papain were different from those from the other cells. On two-dimensional immunoelectrophoresis, the receptor of trypsin-treated cells gave five precipitation lines against anti-stroma and that of papain-treated cells three lines, but any other receptors showed no line. These findings indicate that there are plural receptors for P. coccineus lectin in red cells treated with each of proteolytic enzymes and that the receptors from respective red cells have electrophoretically and serologically different property.


Studies on anti-B specificity and heterogeneity of some invertebrate hemagglutinins. Hemolymph from Charybdis japonica and Lymantria dispar, and saline extract from Eunice kobiensis agglutinated human group B erythrocytes specifically. Multiple agglutinins were identified, and independent agglutinins were separated from hemolymph of Charybdis japonica (anti-Bcj+anti-Xcj) and Lymantria dispar (anti-BLD+RLD) by absorption experiments. By the agglutination and agglutination-inhibition experiments with simple sugars, the specificities of the anti-B agglutinins in these invertebrates were found to differ from each other.


Identification from crossed electro-immunodiffusion patterns of serum. An attempt to identify the individuality from crossed electro-immunodiffusion (CEID) patterns of serum was made by using coefficient of variation (CV), the ratio of standard deviation to mean of ratios of area in couples of corresponding peaks in the CEID patterns. The values of CV in most cases of sera from identical subject were less than 10 except only one case of 10.6. On the contrary, those in most cases of sera from different subjects were more than 20 except a few cases of 14.9 to 20. No overlap of CV values was observed between the two groups. No significant differences were demonstrated in three groups of sex pairing, i.e., male-male, female-female and female-male. From these results, analysis of CEID patterns by employing CV was found to be available for fairly definite identification as well as differentiation but useless for distinction of sexes in cases of different subjects.


Factors influencing the release of renin in patients under chronic dialysis treatment. Plasma renin activities (resting PRA, post-dialysis delta PRA) were studied in 61 patients under chronic dialysis treatment. Removed sodium and removed water were estimated at each dialysis. Delta PRA/removed-sodium and delta PRA/removed-water were calculated as indices in response to the removal of sodium and water during the dialysis. 1)Resting PRA (pre-dialysis PRA) was positively correlated to delta PRA/removed-sodium, delta PRA/removed-water, serum osmolality, and diastolic blood pressure, but negatively to serum sodium concentration, age, and pulse pressure/diastolic blood pressure. Statistically significant factors controlling the resting PRA were delta PRA/removed-sodium, delta PRA//removed-water, and serum sodium concentration. Resting PRA was slightly correlated to diastolic blood pressure and age. 2)Post-dialysis PRA was significantly correlated to the resting PRA, delta PRA/removed-sodium, delta PRA/removed-water, serum sodium concentration, and age, but not to the blood pressure indices.


Diagnosis of early cancer of the colon and rectum. When early cancer of the colon and rectum is defined as in early gastric cancer, i.e., invasion is limited to the mucosa and submucosa, 38 lesions from 35 patients of early cancer of the colon and rectum were detected during the last 20 years. The macroscopic and histologic findings included cancer-containing adenoma in 35 lesions from 32 patients. When the diagnostic methods were compared in 33 cases of early cancer of the colon and rectum, it was found that positive or suspicious cancer was obtained by an x-ray study in 5 of 18 cases examined (28%), by endoscopy in 19 of 31 cases (61%), by cytologic methods in 18 of 21 cases (86%), by biopsy in 19 of 25 cases (76%) and by polypectomy under direct vision through an endoscope in 13 of 14 cases (93%). When both biopsy and cytologic studies were performed in combination with endoscopy (20 cases), either of them was positive in all cases. These results indicate the possibility of correct diagnosis in many cases through the combined use of the cytologic method and biopsy without polypectomy, if the location is previously checked. On the other hand, 104 polyps from 90 patients were removed by means of the snare-electrocautery technique with the use of coagulation current during the last 4 years. The histologic findings revealed cancer-containing adenoma in 13, even in benign appearing polyps and in small polyps 1.0 cm or less in diameter. These results indicate that diagnosis of early cancer of the colon and rectum is difficult without polypectomy, if cancer is not previously suspected.


Severe combined immunodeficiency disease with adenosine deaminase deficiency. Severe combined immunodeficiency disease with adenosine deaminase deficiency was reported of a 1-year-2-month-old boy, who showed clinical picture of pulmonary candidiasis. The serum immunoglobulin level and absolute lymphocyte count were markedly diminished. But immunological study revealed both T- and B-cell surface markers on the peripheral lymphocytes, in spite of complete deficiency of both systems.


Elimination of endrin by mallard ducks.

Elimination of endrin by mallard ducks. Endrin is very toxic to birds and has been implicated in the deaths of birds in nature. However, it is not known how rapidly birds eliminate endrin, a factor important in determining how much is accumulated in tissues. In this study, the loss rate of endrin was followed for 64 days in mallard (Anas platyrhynchos) drakes that had been fed 20 ppm endrin for 13 days. The loss from carcass and blood was described by the equation Y = a e b square root of x where Y = the concentration of endrin in ppm, a = the concentration at day 0, e = the base of natural logarithms, b = the first order rate constant for the elimination process, and x = the number of days after cessation of endrin treatment. Endrin was lost rapidly at first; concentrations in carcasses on a wet-weight basis decreased by 50% in the first 3 days. Thereafter, endrin was eliminated more slowly; elimination of 50% of the remainder required 8.9 days, and it took 32.9 days to lose 90% of the original amount.


Assessment of DNA damage in germ cells of male rabbits treated with isoniazid and procarbazine. The 2 hydrazine derivatives isoniazid (INH) and procarbazine hydrochloride (P) were injected intravenously into rabbits. Radioactive thymidine was injected into both testicles. Rabbits were ejaculated repeatedly, sperms were counted and incorporation of [3H] thymidine into sperm head DNA was determined by liquid scintillation counting. In P-treated rabbits (5 and 50 mg/kg) radioactivity was significantly increased in sperms that were in late phases of spermatocyte and early phases of spermatid maturation at the time of treatment. This indicates that DNA repair synthesis, (unscheduled DNA synthesis, UDS) occurred following drug-induced DNA damage in these germ cells. Normal DNA synthesis in spermatogonia was inhibited by the high dose only. INH (50 and 125 mg/kg) did not cause UDS in spermatocytes and spermatids and did not affect normal DNA synthesis in spermatogonia. The results are in agreement with literature data indicating that P is a potent mutagen and carcinogen. INH, on the other hand, has weak mutagenic and carcinogenic activities that are most apparent in mice.


Effects of low cobalamin diet and chronic cyanide toxicity in baboons. The effects of a low cobalamin (Cbl) diet, together with chronic cyanide or thiocyanate administration in some animals have been investigated in baboons over a period of 42 months. All animals remained healthy throughout the study and gained weight at a similar rate. None became anaemic or showed major haematological changes and there were no major neurological changes. Plasma total Cbl in the animals on the low Cbl diet fell within 9 months to values below the lower limit in man and were lowest at 24 months in baboons not receiving cyanide or thiocyanate. A striking feature in all animals, however, was an apparently seasonal increase in the plasma total Cbl each autumn with a corresponding decrease the following spring. This fluctuation was detected by radioisotopic assay but not by Euglena. Methylmalonic (MMA) excretion after oral valine ranged from 0.1--8.4 mg/24 h and was greatest in animals on the low Cbl diet and not receiving cyanide or thiocyanate. The results suggested an inverse relationship between MMA excretion and plasma total Cbl. Plasma thiocyanate was consistently higher in animals receiving cyanide or thiocyanate and at the end of the study plasma cyanide was highest in animals on the low Cbl diet receiving cyanide. The results support the suggestions that cyanide affects bodily handling of Cbl and that hydroxo-cobalamin plays a part in detoxication of cyanide.


Ninety-day toxicity of photomirex in the male rat. Photomirex (8-monohydromirex) is a demonstrated environmental contaminant and was observed in previous short-term studies to produce lesions in the liver, thyroid and testes of male rats. The present study was undertaken to confirm those observations and to determine the effects after a longer period of exposure. Male rats were fed photomirex for 13 weeks at levels of 0.20, 1.0, 5.0, 25 and 125 ppm in the diet. Deaths were observed in animals receiving the highest dose. Decreased body weight gain and food intake were also observed in that group. Liver weights were increased at 5.0 ppm photomirex and higher. Photomirex caused changes in several biochemical parameters including serum sorbitol dehydrogenase and hepatic aniline hydroxylase activities. Dose-related histological abnormalities were observed in the thyroid and liver starting at the lowest dose level. These results confirm earlier findings and show that photomirex is a potent hepato- and thyrotoxin.


Paraquat poisoning: an analytical toxicologic study of three cases. Paraquat concentrations were measured in tissue, serum, urine and hemodialysate obtained from 3 patients who died 16.5 h, 22 days and 23 days after ingestion. In the patient who died 16.5 h post-ingestion, tissue paraquat levels were high. Kidney and liver had paraquat concentrations of 14 micrograms/g and 13.2 micrograms/g respectively, whereas lung tissue had a paraquat level of 3.8 micrograms/g. Low concentrations of paraquat were detectable in the tissues of the patients who died 22 and 23 days post-ingestion. Early in the poisoning, serum paraquat levels were high and large quantities of paraquat could be removed by both hemodialysis and forced diuresis. During an 8-h period, 713 mg of paraquat were removed by hemodialysis and 340 mg by forced diuresis. After the day of ingestion, little paraquat could be removed by hemodialysis or by forced diuresis; however, at all stages of the poisoning studied, hemodialysis was more effective than forced diuresis in removing paraquat from the blood.


The excretion of trace elements in rat urine after treatment with 2,3-dimercaptopropane sodium sulfonate. Administration of various doses of the chelating agent 2,3-dimercaptopropane sodium sulfonate (Dimaval, DMPS) leads to a greatly enhanced excretion of Zn and Cu. The excretion of Co, Mn, Ni and Fe remains unchanged. The relevance of these findings to the toxicity of DMPS is discussed.


Age-related variations in the testes and prostate of beagle dogs. Variations in the weight and histology of the testes and prostate of Beagle dogs are described. Data were abstracted from records of toxicity studies performed at the Huntingdon Research Centre during the years 1967--77. A sample of 198 dogs aged between 37 weeks and 7.75 years was analysed. When juvenile dogs are studied, a significant proportion show low testicular weight, incomplete spermatogenesis and immature prostate development after 13-week observation periods. The occurrence of these normal variations may complicate the assessment of gonadal toxicity. In other respects, relatively few spontaneous variations from normal are likely to be encountered in these organs after observation periods lasting up to 2 years. 10% of dogs maintained for a 7-year observation period developed testicular neoplasms and a further 8% showed interstitial cell hyperplasia. Epithelial hyperplasia of the ductus epididymis and benign prostate enlargement were also normal features of the aged male Beagle.


Assessment of the PKU card as a retrospective index of neonatal blood lead status. PKU cards were obtained for 50 one-year-old children whose cord blood lead, maternal blood lead and household water lead were known. These 3 values are known to correlate well. Lead values were measured from the blood on the cards, and correlated significantly with the other values. However, predictive validity was poor due to card contamination and it is felt that, for the individual patient, this is not a reliable retrospective index of blood lead at the time of birth.


Distribution and excretion of [14C]citrinin in rats. The distribution and excretion of radioactivity from [14C]citrinin (3 mg/kg, i.v) was determined in male rats. At 0.5 h after administration maximum values of 14.7% and 5.6% of total radioactivity were observed in the liver and kidneys, respectively, and by 6 h decreased to 7.5% in the liver and 4.7% in the kidney. Plasma concentration of 14C decreased from 9.2% at 0.5 h to 4.7% at 6.0 h. 2 plasma elimination rates were observed, with half-lives of 2.6 and 14.9 h, respectively. Approximately 80% of the administered 14C activity was excreted in feces and urine by 24 h after administration. A second group of rats was pretreated with 50 mg/kg of citrinin, i.p., 4 days prior to administration of 3 mg/kg [14C]citrinin, i.v. 30% of the pretreated animals died and the remaining animals were divided into 2 groups on day 4 after pretreatment; rats which were "nephrotoxic" and rats which had "recovered" from the initial insult of citrinin. Proteinuria and glucosuria as well as enhanced urine output were observed in "nephrotoxic" rats 4 days after pretreatment. 24 h after [14C]citrinin, only 13% of 14C activity was detected in the urine of "nephrotoxic" rats. The plasma disappearance curve had 2 elimination rates, with half-lives of 0.6 and 14.1 h. "Nephrotoxic" rats retained 7.5% of the administered radioactivity in the liver compared to 1.3% in the "recovered" rats 24 h after the tracer dose and 47% of the radioactivity was either excreted in feces or in the colon contents after 72 h compared to 17.5% in "recovered" rats. Extraction of urine samples from "nephrotoxic" and "recovered" rats with chloroform suggested increased water soluble metabolites of citrinin in the urine from "nephrotoxic" rats. These data also suggested that in normal rats the kidneys are the major route of elimination of citrinin and its metabolite(s) while in rats rendered nephrotoxic by citrinin pretreatment, elimination is more dependent on hepatic excretion.


Effects of PR toxin on liver cells in culture. We studied the effects on liver cells in culture of PR toxin, a substance produced from Penicillium roqueforti. PR toxin displayed cytotoxicity which increased as a function of its concentration but the form of such toxicity differed, depending on the toxin's concentration. Thus, cells only underwent quick retraction and intensive vacuolization when treated with low drug concentrations, and they came away from the substrate easily under these conditions. By contrast, the major events observed in the case of high concentrations were loss of structure of the nuclei and strong adhesiveness of dead cells to the support. PR toxin already inhibited cell multiplication at low concentrations and became toxic when the concentration was raised; growth inhibition decreased but the toxic effect increased when cells passed from the exponential growth phase to a phase of slower growth. PR toxin inhibited tritiated precursor incorporation into DNA, RNA and proteins in a similar time and concentration-dependent manner. Inhibition of DNA synthesis persisted even after removal of the drug from the medium.


Problems associated with the use of merthiolate as a preservative in anti-lymphocytic globulin. The cytotoxic properties of 2 anti-lymphocytic globulin (ALG) preparations were investigated in vitro by measuring the release of 51Cr from labelled human peripheral blood mononuclear cells, tonsil lymphocytes and Chang cells, incubated with different concentrations of ALG. One of the ALG preparations showed non-selective cytotoxicity in the absence of complement. Evidence was obtained to suggest that this effect was due to merthiolate (sodium ethylmercurithiosalicylate) which had been added to the ALG as a preservative during manufacture. The mercury concentration in the ALG was found to be greater than that stated by the manufacturers. It is conceivable that the clinical use of such as ALG preparation might lead to mercury accumulation in the tissues, with resulting toxic effects. The whole question of the use of merthiolate in the preparation of sera for administration to human subjects needs to be reconsidered.


Oral toxicity of an essential oil from myrtle and adaptive liver stimulation. The acute oral toxicity of an essential oil from the leaves of Myrtus communis (M) in rats was 3.7 ml/kg, and 2.2 ml/kg in mice. After repeated daily doses the toxicity decreased considerably and was calculated to be approx. 6.6 ml/kg after 10 days or 3 weeks pretreatment with M. Daily application of M during 10 days increased the relative liver weight of rats by 10, 18 and 28% at the dose levels of 0.5, 1.0 and 2.0 ml/kg. The hepatic endoplasmic reticulum (microsomal fraction) in the rats increased by 13.5, 24 and 33% after 10 days oral application of 0.5, 1.0 and 2.0 ml/kg of M, respectively. Hepatic cytochrome P-450 started to increase after 0.2 ml/kg, and reached 65% above controls after 10 days application of 2.0 ml/kg. Cytochrome b5 responded in a similar manner. The microsomal N-dealkylation of N-methylaniline increased 2.8-fold after daily administration of 2.0 ml/kg during 10 days. Continuous therapeutic application of M to humans in the magnitude of 1--2 ml daily is considered to be too low to influence the hepatic parameters.


Chronic lead treatment differentially affects dopamine synthesis in various rat brain areas. The effect of chronic dietary lead exposure on brain nigrostriatal, mesolimbic and mesocortial systems was studied. The results show no modification of the dopamine receptors measured either as dopamine sensitive adenylate cyclase or as [3H]spiroperidol binding. On the other hand, dopamine synthesis seems to be reduced in striatum, unaffected in substantia nigra and increased in nucleus accumbens and in the frontal cortex. The increase of DA synthesis observed in some brain areas might be involved in determining the hyperactive behaviour that follows lead intoxication.


Methods for improved light microscope microtomy. Experiments undertaken to improve the ease and quality of preparing tissue for light microscopy have resulted in methods for routine production of high quality serial paraffin and glycol methacrylate sections in the 1 to 4 micrometer range using a standard rotary microtome. These methods involve sectioning with mechanically-broken Ralph-type knives, inspected for quality before mounting with double-stick tape on a holder-fluid trough in which the ribbon floats over an immersed slide and sections are mounted by lowering the fluid level in the trough. Specimen return stroke retraction, necessary for quality sectioning of plastic or with a fluid trough, is inexpensively provided by an attachable specimen holder that fits into the rotary microtome's chuck and, actuated by the microtome mechanisms, retracts the specimen during each return stroke. Descriptions of four instruments: a mechanical breaker for Ralph knives, a knife edge inspection device, an attachable specimen retractor for rotary microtomes, and a section collecting-mounting device as well as details of their use are included. Differences in cutting speed and cutting cycle cadence critical to quality sectioning with different embedments are detailed. Factors important in choosing the best embedment and most appropriate section thickness as well as choosing between dry or wet sectioning methods are also considered.


Transmission and scanning electron microscopy of cell monolayers grown on polymethylpentene coverslips. Plastic coverslips made of polymethylpentene serve as excellent substrates for growth of bovine endothelial cells, and are easily processed for both transmission (TEM) and scanning (SEM) electron microscopy. Portions of the same coverslip (monolayer) are used for both SEM and TEM examination and are fixed, postfixed, and dehydrated as a single entity. The portion of the coverslip for SEM is then excised, critical point dried, and mounted for sputter coating prior to viewing. The remaining piece of coverslip used for TEM is Epon-Araldite embedded, polymerized, separated from the coverslip by liquid nitrogen immersion, and sectioned either "en face" or in cross section for viewing. Coated glass coverslips are not required and organic solvents such as propylene oxide, acetone, and amyl acetate can be used for dehydration and infiltration. Furthermore, specimens do not require re-embedding or blocks to be glued onto blank capsules before sectioning. The number of cells needed to achieve a monolayer is significantly reduced compared to the usual culture flasks, but are abundant enough to assess ultrastructural changes accurately. Support films may be required to prevent folding of the ultrathin section which can obstruct viewing of cells located on the edge of the section.


Modification of a scanning transmission electron microscope specimen holder for large section viewing. A modification of a scanning transmission electron microscope specimen holder which permits full viewing of large plastic embedded tissue sections is discussed. The method for producing one-centimeter diameter "giant" grids is explained and the procedure for sample preparation is outlined. The modification aids the microscopist in his evaluation of tissue structural relationships by providing large areas of tissue for examination and reduces significantly the time required to prepare and examine standard 1-2 mm2 electron microscopy tissue sections. Light and electron microscopic evaluations can be made on the same tissue sections.


Chromosomal banding patterns produced by methyl green-pyronin staining after trypsin treatment. A method is described for producing banding patterns with methyl green-pyronin (MGP) stain in chromosomes of fibrosarcoma cells. 1) The stain was made by mixing equal volumes of 2% aqueous pyronin G, 2% aqueous methyl green, distilled water, and 0.1 M acetate buffer (pH 5.7). 2) Treatment with colcemide and hypotonic KCl (0.075 M) was performed as usual. 3) Metaphase chromosomes were prepared using the flame-drying technique and treated with 0.25% trypsin at 37 C for 45 to 90 seconds. Before staining, the slides were rinsed in PBS, in distilled water, and then were dipped in 0.05 M acetate buffer. 4) Chromosomes were stained for more than 20 minutes, rinsed in distilled water, and hot-air dried. Satisfactory results were obtained in uncontracted metaphase chromosomes. MGP stain has the advantage of permitting much longer trypsin treatment and staining time than the trypsin-Giemsa method while providing satisfactory banding patterns.


A thermally controlled microinjector.

A thermally controlled microinjector. An economical device for 0.1-2 microliter microinjections in animals is described. The expansions of electrically heated mercury is used to provide precise and repeatable injections. The rate of injection can be controlled either manually or by a motor drive.


Remarks about fractionation schemes.

Remarks about fractionation schemes. Today a change from the orthodox "200 rad daily, 5 times per week" scheme to other improved treatment schemes is in the foreground of discussions. However, the situation is very complex and so far no distinct solution has been found. Radiobiology has recommended larger doses at less frequent intervals, but also smaller doses given more often have been used with quite good results. Several factors are important: Tumor conditions, dimensions, growth and sensitivity--reoxygenation of hypoxic tumor cells--recovery and tolerance of normal cells and, last but not least, the general state of the patient. To explain and unravel the problems an example of therapy with high-energy electrons is described. Considering four fundamental successively depending decisions an empirical system is described which may enable us to find improved treatment schemes. Finally a method is explained which makes it possible to transform a treatment scheme to a different equivalent scheme by calculating a total effective standard dose (ESD). This calculation follows exactly the isoeffect principle and makes possible a comparison of different treatment schemes using also different types of radiation.


[Problems of evaluation concerning the success achieved by treating the mastocarcinoma (author's transl)]. The authors analyze the differences of the figures of success communicated in literature for the curative treatment of the mastocarcinoma. They find that neither the therapy method nor the extension of the operated body region are decisive for these figures. The individual success depends on the stage, and the group result is only influenced by the composition of the patients; These observations explain also why the figures of success of the curative therapy of the mastocarcinoma which were based on the stages of the disease have been at a standstill since 25 years. A decisive improvement, however, can be anticipated by virtue of an intensified prevention of mastocarcinoma and a changement of the therapy conception including a prophylaxis against metastases.


[Experiences with orthovoltage therapy in cancer of the breast (author's transl)]. After 22 years of sole orthovoltage therapy the results and experiences arrived at in the course of treatment are subjected to a final review. The results are compared to those of other authors, obtained with megavoltage therapy, and a critical analysis of the problems existing in the radiation therapy of cancer of the breast is presented.


A simple formula for depth dose calculation for Co-60 teletherapy beam dosimetry. Knowledge of dose at all the points of interest, in the plane of tumour, is essential for treatment planning. A very simple formula for scatter dose calculation along the central axis of a Co-60 beam has been derived. This formula uses primary dose at depth d, scatter air ratio at the depth of maximum ionisation and the effective depth of the volume, irradiating the medium. The method for calculation of percentage depth dose at any point in the principal plane has been explained in detail. The simple form of the formulation will help in improving the treatment plans for treatments of lesions using Co-60 teletherapy machines.


[About the present state of technology of therapy simulators: comparison of the characteristic data of 8 devices (author's transl)]. The author compares properties and characteristics data of therapy simulators of different producers (AECL, Oldelft, Philips, Picker, Santax, Siemens, Toshiba), outline their value as against the computer tomograph, and mention the actual trends of development.


Absorbed dose rate in tissue around uniformly loaded linear radium and 137Cs-sources encapsulated in platinum. A method is described for calculating the absorbed dose rate in tissue around encapsulated linear radium- or 137Cs-sources. A variable factor is derived relating this quantity to the absorbed dose rate from the unshielded source in air. This factor depends on a set of geometrical parameters including some data concerning the shape of the source ends.


Microdosimetric measurements of ionization by some radioisotopes. Microdosimetry has been a fast growing field in dosimetry during the last years. We have measured and calculated microdosimetric quantities which are important in correlation between the energy deposited in microscopic volumes and the biological effect. We have measured the energy distribution spectra in volumes with diameters of 1 and 2 mum and calculated the dose mean of specific energy and the mean event number per unit absorbed dose for the radioisotopes: 60Co, 137Cs, 99Tcm, 125J and the beta emitter 90Sr.


Kinetics of regrowth of V79 spheroids after gamma- and pi--irradiation. Spheroids of V79 cells were irradiated with 180 MeV/c pi--mesons in the peak region and with gamma-rays. Doses of 900 and 1500 rad were applied, respectively, yielding 14% surviving cells. Regrowth of the spheroids was followed by measuring volume changes, cell multiplication as well as viability and proliferation of cycling and resting spheroid cells. While volume growth after irradiation was equal for both radiations, the time profiles of cell density, viability and mean DNA content per cell were found to be different. Consistently, a more prompt and pronounced response to pion irradiation could be inferred and was discussed in terms of LET dependent proliferation and elimination of inactivated cells and their subsequent substitution by viable progeny cells. Recruitment of resting cells was observed with both radiations.


The effect of dianabol on certain cell energy processes in postirradiation disease. III. The effect of chronic administration of dianabol and irradiation on oxidative phosphorylation in rat liver mitochondria. Chronic administration of Dianabol did not prevent the radiation-induced changes of oxidative phosphorylation in rat liver mitochondria. Irradiation of rats with X-rays, in a dose of 600 R for the whole body, causes damage of oxidative phosphorylation in rat liver mitochondria. Progressive reducation of oxygen consumption, decrease of high-energy bond formation and a drop of the P/O coefficient were observed in irradiated animals. The presumable role of the postirradiation damage of oxidative phosphorylation and the effects of radioprotective compounds on this process are discussed.


[Results from 215 patients with laryngeal carcinoma treated with sole radiotherapy or with a combination of radiotherapy and operation (author's transl)]. Results of radiotherapy alone or in combination with surgery in 215 patients with laryngeal cancer are reported (treatment time between 1963 and 1976). In patients with glottic cancer, the cure rate is about 80% and surgical treatment for persistent/recurrent cancer was necessary only in a few cases. Analysis of tumor dose, tumor control rate and complication rate shows that in cases with glottic cancer a tumor dose between 1900 and 2000 ret is necessary and tolerable. In patients with supraglottic laryngeal cancer, the cure rate is about 55%. Early stages (T1N0M0) were mostly treated by radiotherapy alone, moderately advanced stages (T1N1,T2N0+1) mostly received radiotherapy as a primary treatment and were operated in case of irradiation failure. Advanced stages were primarily treated by a combined therapy, and by radiotherapy alone if they were inoperable. In early cases, who are to be treated by radiotherapy alone, the tumor dose should be about 1900 ret. In moderate cases, treatment by primary irradiation alone is possible, if the patient can be followed up regularly; surgery is indicated, if the patient cannot unequivocally be classified as symptomless. In some of these cases, there is no tumor demonstrable histologically in the excised larynx. In using this treatment policy, there is a better functional treatment result than in using primary combined treatment in moderately advanced cases with supraglottic laryngeal cancer.


[Cancer prophylaxis: gain or risk? (author's transl)]. One of the most important measures for the improvement in curative cancer therapy, stagnant during 25 years, will be a cancer prophylaxis as extensive, intensified and directed as possible. Seventy percent of all the cancerous diseases are produced by seven cancer species. Of these seven species, four are appropriate for a promising cancer prophylaxis: the mammary, uterine and intestinal catzinoma in women, the intestinal carcinoma in men. Risks as well as improvements concerning prophylaxic examination for cancer of the breast or prostatic carcinoma are discussed in detail.


Calculation of the dose distribution around 137Cs tubes used in clinical brachytherapy. The dose distribution in water around a CDC.J 137Cs tube is calculated with a computer. In the transverse direction the calculated dose rates showed to be almost unsusceptible for the value of the platinum absorption coefficient. In the longitudinal direction, however, the value of the linear absorption coefficient plays a more prominent role. An effective absorption coefficient for the filter material was determined by measurement, using a photographic film. This was done by relating the filmdensity along the source axis with the filmdensity along the transverse axis, where the dose rate is well known. In this way an effective linear absorption coefficient for platinum was obtained which gives a good agreement between the experimental and theoretical results. For use in a computer program for clinical use, the dose distribution around a cesium tube has been calculated in a specific spherical coordinate system using 101 radii.


Multidetector endodosimetry probe with silicon-pn-junction-diodes for in-vivo-dosimetry. In-vivo-dosimetry is necessary to prevent overdosage during radiation treatment techniques of great risk. New special dosemeter probes with up to five Si-pn-junction-diodes in a silicone tube were developed and tested for endodosimetry. They show high efficiency and good linearity of response, without dose-rate dependence. The steep increase of the sensitivity for low energies causes a phantom depth dependence of response. A low temperature dependence but a large anisotropy of response is found for the tested diodes. These multidetector probes proved to be sufficient for direct reading endodosimetric control of dose and dose-distribution during radiotherapy.


[Theoretical derivation of a logarithmic equivalence function for the fractionation in depth therapy (author's transl)]. The effect of fractionated radiation therapy is essentially dependent on recovery of the tissues during the fractionation intervals. Supposing the recuperative capacity of irradiated tissues is lessening in the course of a depth therapy series (introduction of the term "change of the seeming recovery"), an equivalence function for fractionation is obtained and the conventional logarithmic function therefrom derived. Qualities of the here developed equivalence function are discussed.


Participation of granulosa cell populations in radiation response of the follicular apparatus in the ovary of mice. The aim of this paper was to investigate the fates of granulosa cells in three different types of the follicles during 10 hours after single irradiation with 100 resp. 200 R. Using flash labelling with 3H-thymidine, observations of the ovarian anatomical structures during 4--5 estruses after 150 R were made. It has been found, that the basic radiobiological event, influencing further fates of the follicles, is delay of granulosa cell proliferation. Consequently, the further growth and maturation of the follicles is retarded and a reduction of ovulation, a lengthening and an irregularity of estrus take place. The killing effect within granulosa cells is not significant and is of secondary importance.


Radiation effects on the parotid gland of mammals. Part 1: Modifications on the morphology and uptake and elimination after 14C-leucine administration. U-14C-Leucine uptake and eliminaton in connection with morphological investigations after sublethal doses have been studied. The tracer uptake and elimination curves do not show significant differences between controls and irradiated animals, except for early appearance of initial peaks. An increased uptake of labeled amino acid was observed only 32 and 120 hours after irradiation, when sacrifices were done at the same hours of the day. The dose employed does not seem to affect in any significant way either the acinar and duct cells or other structures. Only a mild thickening of the connective tissue has been observed in the parotid of animals sacrificed at longer intervals.


The reduction of 5 alpha-cholestan-3-one and 5 beta-cholestan-3-one by some boranes and hydroborates. This paper describes the high yield synthesis of 5 alpha-cholestan-3 alpha-ol and 5 beta-cholestan-3 beta-ol by the reduction of 5 alpha- and 5 beta-cholestan-3-one using using potassium and lithium-tri-sec-butyl-hydroborates as reducing reagents. Attempts to obtain the same alcohols using other bulky boranes resulted in the equatorial products.


Metabolism and binding in vitro of 5 alpha-androstane-3 beta, 17 beta-diol and of 5 alpha-androstane-3 alpha, 17 beta-diol in cell fractions of rat ventral prostate and liver. Rat ventral prostate and liver were investigated for the binding in vitro to particulate fractions and for the metabolism of 5 alpha-androstane-3 beta, 17 beta-diol. Comparative investigations were carried out on the metabolism of 5 alpha-androstane-3 alpha, 17 beta-diol. Preparations of the liver were investigated in order to establish the organ specificity of the method. In the prostate, the bulk of the metabolites of 5 alpha-androstane-3 beta, 17 beta-diol was present as steroids of high polarity. Of the less polar metabolites, 17 beta-hydroxy-5 alpha-androstan-3-one, 3 beta-hydroxy-5 alpha-androstan, 17-one and 5 alpha-androstane-3 alpha, 17 beta-diol were detectable. The binding of a 5 alpha-androstane-3 beta, 17 beta-diol to mitochondria and microsomes was unspecific. In the liver, among the less polar metabolites, 3 beta-hydroxy-5 alpha-androstan-17-one was the main metabolite, and the binding was unspecific. The main metabolite in the prostate homogenate of 5 alpha-androstane-3 alpha, 17 beta-diol was 17 beta-hydroxy-5 alpha-androstan-3-one. The portion of highly polar steroids was very low. The portion of unmetabolized hormone was distributed almost equally among the different cell preparations except the nuclei, in which 17 beta-hydroxy-5 alpha-androstan-3-one was higher and 5 alpha-androstane-3 alpha, 17 beta-diol was lower than in the remaining cell fractions.


BOMT (6 alpha-bromo-17 beta-hydroxy-17 alpha-methyl-4-oxa-5 alpha-androstan-3-one) is not an androgen antagonist within the central nervous system. This study investigates the efficiency of BOMT as an androgen antagonist within the central nervous system. The efficiency of BOMT in suppressing neural receptor binding of testosterone, and the ability of this antiandrogen to block the feedback loop of testosterone onto the central nervous system, as evidenced by plasma testosterone levels, is reported. BOMT was found to be unable to open the feedback loop of testosterone onto the central nervous system, which was correlated with the low competing efficiency of this antiandrogen for receptor sites in vitro within the hypothalamic-preoptic area of the brain - a region known to be involved in gonadotrophin secretion. The observed divergence in the degree of antiandrogenicity of BOMT between peripheral and central target tissues of testosterone is discussed.


Effects of various doses of testosterone propionate on intratesticular and plasma testosterone levels and maintenance of spermatogenesis in adult hypophysectomized rats. Adult hypophysectomized rats were maintained on different regimens of testosterone propionate (TP) treatment for 27 days (0.2, 0.4, 0.6 and 1 mg/day) and autopsied 16 hours after the last injection. Blood samples were taken, sex organs were weighed and one testis from each animal was fixed in Bouins fluid for histologic analysis. The other testis and blood were used for testosterone (T) determinations. Both testicular and plasma T were below detectable levels in hypophysectomized control rats. The plasma T level showed a dose response relationship with increasing dose of TP but such was not the case for intratesticular T concentrations. Qualitative and quantitative evaluation of testis sections showed that spermatogenesis was incomplete in rats receiving 0.2 mg TP/day characterized by the absence of step 15 to 19 spermatids, degeneration of some pachytene spermatocytes and a significantly lower yield of B type spermatogonia. Analysis of testis sections from animals treated with 0.4 to 1 mg TP/day showed complete maintenance and maturation of pachytene spermatocytes, meiosis and spermiogenesis. However, even with the highest dose of TP (1 mg/day) the total yield of B type spermatogonia was only about 58% of the intact controls. It is concluded that at least 0.4 mg/day of exogenous TP is essential for qualitative maintenance of spermatogenesis in hypophysectomized rats with an intratesticular T concentration of 17 to 18 ng/gm testis.


Side chain-hydroxylated sterols of the red alga Asparagopsis armata: significant products or artifacts due to autoxidation? Sterols characterized by an allylic hydroxyl group in the side chain, such as stigmasta-5,28-diene-3 beta, 24 epsilon-diol (1), cholesta-5,23-diene-3 beta,25-diol (2) and cholesta-5,25-diene-3 beta,24 epsilon-diol (3), have been identified several times in various marine algae. Their origin was considered as doubtful: they could have been bona fide constituents of the alga, or be artifacts caused by autoxidation during the isolation process. We have shown that the dihydroxy steroids 2 and 3 can indeed be produced by the autoxidation of cholesta-5,24-dien-3 beta-ol (desmosterol) (5), but that they are nevertheless present in the taxonomic significance.


The synthesis of diazo, halo, and sulfoxy bile acid derivatives: potential affinity labels. Bile acid derivatives, with and without C-3 sulfate groups, and having either the diazo- or halomethylketone moieties, have been synthesized in good yield and purity. The synthetic sequence, COOH leads to COC1 leads to COCHN2 leads to COCH2X, was used with deoxycholic and cholic acids, which requires carefully controlled quench, work-up, and purification procedures, especially for the 3-sulfate esters (made from deoxycholic acid derivatives only). The pure title compounds are anticipated to be useful chemical probes (affinity labels), especially the completely water soluble sulfates, toward our studies of ileal active transport of bile salts. A new use for Sephadex LH-20 as a sulfate ester protecting group is reported. Also developed were the use of acetamide hydrochloride complex as a mild hydrochlorination reagent and a neutral desalting method for sulfate esters of deoxycholic acid derivatives.


Synthesis of 17 beta-[(1S)-1-hydroxy-2-propynyl]- and 17 beta-[(1R)-1-hydroxy-2-propynyl]androst-4-en-3-one. Potential suicide substrates of 20 alpha- and 20 beta-hydroxysteroid dehydrogenases. The title compounds have been synthesized for evaluation as potential suicide substrates of 20 alpha- and 20 beta-hydroxysteroid dehydrogenases. Synthesis was achieved by the following route. Acetylenedimagnesium bromide was reacted with 3 beta-hydroxyandrost-4-ene-17 beta-carboxaldehyde to give 17 beta-[(1R,S)-1-hydroxy-2-propynyl] androst-4-en-3 beta-ol. Separation of the R and S diols was achieved by HPLC (high pressure liquid chromatography). Selective oxidation of the 3 beta-hydroxyl group with Jones reagent at 0 degrees gave the title compounds. Further oxidation with Jones reagent converted each acetylenic alcohol to the conjugated acetylenic ketone, 17 beta-(1-oxo-2-propynyl)androst-4-en-3-one.


Estrogen metabolism in neural tissues of rabbits: 17 beta - hydroxysteroid oxidoreductase activity. Incubation of adult rabbit neural tissue homogenates with (3H)-estrone and (3H)-estradiol revealed that the conversion of estrone to estradiol is higher in both male and female animals than estradiol to estrone. Both 17 oxidation and reduction are higher in male animals than in females. However, it is observed that the quotient of estrone leads to estradiol/estradiol leads to estrone for pituitary tissue and hypothalamus are higher in females than in males. There is no such dimorphism in cerebral cortex. The overall metabolism in pituitary is higher than in hypothalamus and cortex in both sexes. These results suggest that 17 beta hydroxysteroid oxidoreductase activity may play an important role in the regulation of estrogen function in neuroendocrine tissues.


Relationship between estrogen receptors, 17 beta-hydroxysteroid dehydrogenase and estrogen content in human breast cancer. Estrone and estradiol levels in tumor tissue cytosols were determined in 11 premenopausal and 20 postmenopausal women at the same time that 17 beta-hydroxysteroid dehydrogenase and estrogen receptors (ER) were carried out on their breast cancers. Estrogen receptor positive tumors showed significantly higher levels of estrone and estradiol. However, all ER negative tumors contained measurable amounts of both estradiol and estrone. Higher levels of estrone were observed in ER negative tumors which correlates well with high 17 beta-hydroxysteroid dehydrogenase activity. These results suggest that false negative receptor assays in the premenopausal women is not likely to be due to occupancy of receptors by endogenous estrogens. Furthermore, the higher estrone content in the ER negative group is probably due to high 17 beta-hydroxysteroid dehydrogenase activity inherent to these tumor cells.


An evaluation of systems for paper chromatography of C21 steroids. A group of moderately polar C21 steroids (3-5 oxygen functions) has been chromatographed in 9 solvent systems. Using the concept that standard deviation of the mean RF is an index of chromatographic resolution, and that coefficients of correlation between sets of RF data can be used to quantify the similarities of chromatographic systems, we have evaluated the resolving properties of the systems when used individually, and also when used in combinations of two, three and four. The discriminating powers of some of the most effective individual systems, and some of the sequences of systems which are most efficient, are shown graphically as chromatography trees. The relationship between the total effective standard deviation of a group of systems which are used in sequence and the probability that a pair of compounds will be separated by more than 0.10 RF is discussed.


Pituitary metabolism of 5alpha-androstane-3beta-17beta-diol: intense and rapid conversion into 5alpha-androstane-3beta,6alpha,17beta-triol and 5alpha-androstane-3beta,7alpha, 17beta-triol. In the male rat pituitary, 5alpha-androstane-3beta, 17beta-diol (3beta-diol) is extensively metabolized into polar steroids. They were identified as 5alpha-androstane-3beta, 6alpha-17beta-triol (6alpha-triol) and 5alpha-androstane-3beta, 7alpha, 17beta-triol (7alpha-triol). 6-alpha-Triol represents 53% and 7alpha-Triol 28% of the total 3beta-diol metabolites. The remaining percentage is related to 6beta and 7beta isomers. The biological role of triols is still unknown.


Preparation of [3beta-3H] labeled bile acids and bile alcohols. [3beta-3H]-bile acids and bile alcohols may be useful for metabolic studies in man and animals because the 3-position is invulnerable to bacterial attack. A number of tritium labeled bile acids and bile alcohols were prepared by selective oxidation of the hydroxyl group at carbon-3 followed by reduction with NaBT4. In each case, the bile acids and bile alcohols epimeric at carbon-3 were resolved by analytical and preparative thin-layer chromatography and characterized by gasliquid chromatography. The average yield was 60--65% and specific activities of the final products were in the range of 7.4 x 10(7) dpm/mg.


Cholesterol biosynthesis and progesterone production by chorionic cells of the early sheep conceptus in vitro. Dissociated trophoblast cells from the chorion of the 21-day sheep conceptus were shown to produce progesterone in vitro. Evidence that this represented synthesis de novo was provided by the prolonged production of progesterone in the absence of added precursor steroids. Radioactive cholesterol was isolated after incubation of dispersed chorionic cells with acetate-1-14C.


Percentage binding of testosterone and dihydrotestosterone and unbound testosterone and dihydrotestosterone in rabbit maternal and fetal plasma during sexual organogenesis. The percentages of bound testosterone (17 beta-hydroxy-4-androsten-3-one; T) and dihydrotestosterone (17 beta-hydroxy-5 alpha-androstan-3-one; DHT) and their unbound concentrations were determined in pregnant rabbits and their fetuses from the 18th day of gestation to birth. T and DHT were also measured in fetal testes. In the testis, the total T/total DHT ratio, very high at 22 days (73.7 +/- 15.2), decreased until birth (6.7 +/- 0.8). In male fetuses the concentrations of total and unbound circulating T and DHT were always low and did not show any peak during sexual organogenesis. The percent binding of T (from 73.0 +/- 0.5 to 77.6 +/- 0.6) and DHT (from 76.5 to 83.7 +/- 1.1) in fetuses were similar in both sexes and significantly lower than those measured in mothers (T: from 87.2 +/- 0.6 to 91.6 +/- 0.9; DHT: from 87.3 +/- 0.9 to 93.8 +/- 0.9).


Comparative effects of canrenoate-K and prorenoate-K upon aldosterone biosynthesis in perifused frog interrenal glands. To investigate the possible direct effect of two aldosterone antagonists (Canrenoate-K and Prorenoate-K) upon mineralocorticoid biosynthesis a perifusion system technique has been developed. Frog interrenal tissue was selected for its ability to secrete huge amounts of aldosterone (twice as much as corticosterone in resting conditions). Throughout the experiment, secretion of aldosterone was measured every ten minutes by means of a sensitive and highly specific radioimmunoassay method. Increasing concentrations of both Canrenoate-K and Prorenoate-K (ranging from 10(-4)M to 10(-3)M) caused a dose-related inhibition of aldosterone output. At a dose of 3.16 x 10(-4)M, Prorenoate-K appeared to be somewhat more potent (57.8% inhibition) than Canrenoate-K (47.8% inhibition). Infusion of both Canrenoate-K and Prorenoate-K at a dose of 5 x 10(-4)M during 1 or 2 hours induced a similar sharp decrease in mineralocorticoid secretion. Thus, it appears that Canrenoate-K and Prorenoate-K beside their well known effects at renal tubular receptor sites do also inhibit aldosterone biosynthesis. These results indicate that in vivo administration of aldosterone antagonists may first involve a transient decrease in aldosterone secretion. Furthermore, they suggest that mineralocorticoid biosynthesis might be regulated by a short loop feedback mechanism.


The ecdysteroids from the tobacco hornworm during pupal-adult development five days after peak titer of molting hormone activity. Six naturally occurring C27 ecdysteroids were isolated and identified from the tobacco hornworm during pupal-adult development five days after peak titer of molting hormone activity. In order of decreasing quantities the hormones were: 20,26-dihydroxyecdysone, 3-epi-20-hydroxyecdysone, 20hydroxyecdysone, 3-epi-20,26-dihydroxyecdysone, 3-epi-ecdysone, and ecdysone. 20-Hydroxyecdysone, in an earlier study, was the major molting hormone present at peak titer during pupal-adult development. The major ecdysteroid present during embryonic development in this insect, 26-hydroxyecdysone, was not detected. The copresence of all six of these ecdysteroids from a single developmental stage of an insect provides information on the metabolic interrelationships that exist among these steroids and on their possible function(s) in insects. The 3alpha-ecdysteroids were far less active than the 3 beta-epimers in the house fly assay. The significance of epimerization is discussed.


Synthesis and some reactions of 6-bromoandrogens: potential affinity ligand and inactivator of estrogen synthetase. The synthesis of epimeric 6-bromo-4-androstene-3,17-dione (1a and 1b), 6-bromotestosterone (2a and 2b) and its acetate (3a and 3b), and 6-bromo-16 alpha-acetoxy-4-androstene-3,17-dione (5a and 5b), and 6 beta-bromo-16 alpha-hydroxy-4-androstene-3,17-dione (4) is described. The interconversions among compounds 1, 2, and 3 are also studied. The 6 beta-isomer (1b, 2b, and 3b) was epimerized to the 6 alpha-isomer (1a, 2a and 3a) in carbon tetrachloride or chloroform-methanol (9:1) and the 6 alpha-isomer was isolated by fractional crystallization from the epimeric mixture. 6 alpha-Bromo isomer 1a was also epimerized back to 6 beta-bromo isomer 1b in chloroform-methanol (9:1). Two polymorphic forms of 6 beta-bromotestosterone acetate (3b) were isolated (mp. 114--117 degrees and 138--141 degrees). The 6 beta-bromo isomers were found to be unstable in methanol and decomposed to give 5 alpha-androstane-3,6-dione derivative (6). The results of irreversible inactivation of human placental androgen aromatase with some of these 6-bromoandrogens are discussed.


Selective catalytic reduction of 7-methyl-6-dehydrotestosterone acetate to 7 beta-methyltestosterone acetate by benzyl alcohol. Using benzyl alcohol as a hydrogen donor in the presence of Pd on charcoal, 7-methyl-6-dehydrotestosterone acetate was selectively reduced to 7 beta-methyltestosterone acetate in 90% yield. The addition of hydrogen atoms to the 6, 7 double bond proceeded from the less hindered alpha-face of the steroid molecule, giving rise to the 7 beta-methyl product. Gas chromatograph analysis indicated small amounts of the 7 alpha-methyl epimer, 7 beta-methyl-5 alpha-dihydrotestosterone acetate and the 5 beta-epimer. The 6,7 double bond was hydrogenated in preference to 4,5 double bond, although both are trisubstituted.


Damage to the biliary tract during preservation. Biliary complications following liver transplantation continue to present major difficulties. Damage to the biliary tract has been reported after preservation of the liver before transplantation. Studies in the rabbit are reported that confirm the extensive biliary autolysis during preservation. This damage is directly related to the presence of bile in the biliary tract and can be reduced by the washout of bile and biliary perfusion.


Effect of ureteral ligation and nephrectomy on granulocyte-macrophage progenitor cells and azathioprine toxicity. Using an in vitro quantitative clonal culture technique of bone marrow granulocyte-macrophage progenitor cells (colony-forming units culture (CFU-c)), we studied the hematopoietic toxicity of azathioprine after unilateral and bilateral ureteral ligation, unilateral and bilateral nephrectomy, and splenectomy in C57BL/6 mice. Analysis of femoral bone marrow 18 hr after i.p. injection of azathioprine (300 mg/m2) revealed increased CFU-c toxicity in comparison to controls as follows: (1) bilateral ureteral ligation, P less than 0.01; (2) bilateral nephrectomy, P less than 0.01; (3) unilateral ureteral ligation, P greater than 0.05 less than 0.1; (4) unilateral nephrectomy, P, not significant; and (5) splenectomy, P, not significant. Extrapolation from a dose-response curve for the toxicity of azathioprine on the bone marrow CFU-c indicated that bilateral ureteral ligation and bilateral nephrectomy had the effect of a 25 to 50% increase in the azathioprine dose. After bilateral ureteral ligation, serum granulocyte-macrophage colony-stimulating factor levels were increased and in vitro tritiated thymidine suicide studies showed an increased proliferative rate of the CFU-c. Since azathioprine is a predominantly cell cycle-specific agent, we suggest that increased sensitivity to azathioprine is related to the increased proliferative rate of the CFU-c. The findings provide a rationale for a clinical policy of azathioprine reduction when there is depressed renal function.


Serial quantitation of in vitro lymphocyte responsiveness to concanavalin A. Eight normal donors were serially tested over a period of 1 to 7 months for their in vitro lymphocyte responsiveness (IVLR) to 50 microgram of concanavalin A (Con A) per ml. 3H-Thymidine incorporation was monitored by routine scintillation counting and compared to actual numbers of proliferating cells (lymphoblasts) as measured by laser (flow) cytometry. Determination of total cell numbers or lymphoblast numbers was found to be superior (P less than 0.0001 and P less than 0.001, respectively) to 3H-thymidine incorporation as a measure of serial stability.


In vitro generation of primary cytotoxic lymphocytes against L5178Y leukemia antigenically altered by 5-(3,3'-dimethyl-1-triazeno)-imidazole-4-carboxamide in vivo. Drug-mediated tumor antigens (DMTA), which appear after in vivo treatment of murine lymphomas with 5-(3,3'-dimethyl-1-triazeno)-imidazole-4-carboxamide (DTIC), are shown to elicit strong primary cell-mediated responses in vitro. Specific cytotoxic response is obtained using CD2F1 (H-2d/H-2d) spleen cells cocultured in vitro for 5 days with inactivated histocompatible L5178Y/DTIC lymphoma cells in a one-way mixed lymphocyte tumor culture. Sensitized lymphoid cells are markedly cytotoxic for L5178Y/DTIC targets and are minimally or not active against the parental lymphoma. Marginal or no cross-reactivity is found between L5178Y/DTIC lymphoma and other untreated or DTIC-treated tumor or normal cells. Cytotoxic activity is T dependent, since it is abrogated by treatment in vitro with anti-serum and complement and is not elicited by spleen cells of athymic (nude) donors.


[Nuclear pore density in rat liver cells during regeneration and x-ray irradiation of the animals]. Cytoplasmic as well as nucleoplasmic surfaces of the pore complexes (PC) could be observed using freeze-etching method. The density of PCs per 1 micron2 of nuclear envelope (NE) surface in regenerating liver (9.9) is twice as that in resting liver (5.3). 1 hour after 1200 R X-ray irradiation the pore density in regenerating liver decreases 5.8-fold, consisting only of 1.7 PCs per 1 micron2 of the NE. The structure of the PC after irradiation undergoes degradation and normal PCs practically disappear; only their "ghosts" remain. Peripheral and possibly central granules of the PC appear to consist of some subunits with their diameter of 4--5 nm. The central granule forms a channel through which RNA containing material may be transported from the nucleus to the cytoplasm. The non-uniform state of the PC, observed on platinum-carbon replicas of cleaved nuclei, and the non-altered PC associate with the dense lamina of the NE, after detergent treatment of isolated nuclei indicate that the PC could be formed inside the nuclei and to be "inserted" into the NE membranes in the course of their processing.


[Ultrastructure of the specialized intercellular contacts in an L-cell culture treated with low concentrations of lanthanum]. The electron microscope study of L-cells treated with 1 mM lanthanum has shown an increased amount of gap-like junctions and a decreased number of tight junctions compared to the norm. No damage in the plasma membrane structure and other cell organoids was observed. This may suggest that following the above treatment, the amplified intercellular exchange is associated with the increase in the quantity of intercellular highly permeable junctions. In this case lanthanum may serve as a membrane structure modificator. This effect must be taken into account when the electron microscopists use the lanthanum label for studying specialized intercellular junctions.


[Relationship of the DNA content in the nuclei of Amoeba proteus to the food regimen]. The relative DNA content of isolated Amoeba proteus nuclei has been measured by cytofluorometry. With the amoeba strain studied, the generation time is roughly equal to 48 hours at 25 degrees C, and with the presence of food in the medium. After the synchronous divisions, amoebae were maintained in the medium either with or without food organisms (Tetrahymena pyriformis). DNA contents in the nuclei of both the amoebae groups were measured within 4 and 48 hours after division. Before 16 hours, the nuclear DNA contents did not differ in either group. Starting from 20 hours, the DNA amount in fed amoebae exceeded that in starved animals. On the whole, the differences in DNA quantity increased by a 48th hour after division, when the nuclei of the former contained 145% DNA of the latter. The results obtained suggest that the DNA synthesis in amoeba nuclei may proceed during the whole interphase, and that during the second half of interphase the content of DNA may depend on the feeding intensity in amoebae. After refeeding the starved animals, DNA contents in their nuclei increased to reach the same level as in the constantly fed amoebae seen in the end of interphase.


[Intranuclear development of the macrogametes of the coccidian, Tyzzeria parvula]. The fine structure of macrogametes of a goose coccidium Tyzzeria parvula has been studied, intranuclear localization of these being discovered. Unlike other coccidia, macrogametes of T. parvula display only one type of wall-forming bodies. Deep invaginations are formed on the surface of macrogametes, in which fragments of host cell nucleoplasm, separated with the membrane of parasitophorous vacuole, are invaginating. They may be connected with process of parasite's feeding.


[Karyotypic variability of a tumor cell population in the process of prolonged growth in the body]. An ascite variant of spontaneous mammary gland tumor of C3H/Sn mice was transplanted subcutaneously into syngenic mice. During the tumor growth (150 days) some alterations occur in the genetical structure of the tumor population: a change in the stem line, the increase and a further decrease of nearoctaploid cells.


[Method of lysing animal cells grown on slides on top of an alkaline saccharose gradient]. A device is described providing for lysis of cells grown on glass slides, directly on the top of sucrose gradient. The device allows to avoid cell removing with trypsin, EDTA or due to scraping. The possibility to use this method in studying DNA damage and repair processes in cultured animal cells has been shown using diploid human fibroblasts.


Descriptive epidemiology of Hodgkin's disease in the province of Torino, 1965-71. Age-standardized incidence rates of Hodgkin's disease in the province of Torino in 1965-1971 were 2.81/100,000/year in males and 1.64 in females, i.e., somewhat lower than in urban areas of other industrialized countries. In the whole province, age-distribution for males was similar to previous findings elsewhere, with a young-adult peak at ages 25-34. However, in the city of Torino, males showed a first peak in the age-class 35-44, while in the rest of the province an infantile peak (0-14 years) was found. By dividing the latter area in suburbs and nonmetropolitan area, the infantile peak was confirmed only in the nonmetropolitan area. Females showed a different behavior: in the city of Torino there was a unimodal curve (peak in age-class 55-64), and in the rest of the province there was a bimodal curve with a first peak in age-class 15-24. These patterns have been analyzed in relation to the suggestion of a distribution by age of Hodgkin's disease following 3 types (nonindustrialized countries, urban areas of industrialized countries and rural areas of industrialized countries). No significant differences in the distribution of cases were found among the smallest geographical units considered (ULSs), in contrast with previous findings on cancer of the larynx and bladder, which showed marked geographical differences within the province of Torino.


Time dependence of hematoporphyrin distribution in selected tissues of normal rats and in ascites hepatoma. The distribution of hematoporphyrin was determined in normal rats and in rats bearing ascites hepatoma as a function of time after i.p. injection of 10-20 mg/kg of dye. In both cases, hematoporphyrin displayed a high affinity for the tumor cells. At 20 mg/kg, the maximum difference between the amount of hematoporphyrin accumulated in the tumor and in the liver was obtained at 12 h after injection (tumor/liver ratio = 28). Our results suggest that hematoporphyrin is almost exclusively metabolized in the liver and excreted via the biliary tract, whereas only minor amounts are metabolized in the tumor cells. Moreover, the binding between the porphyrin and tumor cells is competitive with serum protein binding.


Clinically occult breast lesions: criteria of radiological diagnosis as a guide to the choice of biopsy technique. A series of 90 clinically occult operated breast lesions, diagnosed in the course of mammographic mass screening for early detection of breast cancer, is reported. Five radiological patterns of suspect lesions are identified and compared with histological diagnosis of the biopsy. Diagnostic quadrantectomy is suggested for clinically unapparent lesions showing a radiologic pattern which has a high probability of finding a cancer or preneoplastic lesion (severe epitheliosis). Alternative biopsy methods to diagnostic quadrantectomy are recommended for lesions showing other radiological patterns, allowing for a more limited breast tissue excision.


Evaluation of the scintigraphic technique for detecting bone metastases in 100 breast cancer patients. In an attempt to evaluate the utility of bone scintigraphy for the diagnosis and follow-up of skeletal metastases, radioisotopic scan (total body) by 99mTc has been performed on 100 patients suffering from breast cancer in various stages. The radiographic control followed the scan on the basis of clinical or scintigraphic suspicion of bone lesions. Twenty-one patients showed metastases by scan, and in all but 2 the radiography was concordantly positive; in 22 subjects scintigraphy and radiology demonstrated non-malignant lesions. Both scan and radiology were used to explore 169 metastatic localizations, with concordant results in 45% of the sites; however, 7% of the sites with discordant results became positive for metastasis by radiology some months after the scan positivity. The false-negative results were more frequent in the radiographic study (27%) than with the scintigraphic scan (21%). The latter technique was unable to detect osteoblastic lesions in particular. The scintigraphic test also gave reliable results in the follow-up procedure, frequently anticipating the radiological modifications of metastatic lesions. In conclusion, bone scan often appears to be more accurate and preocious than the roentgengraphic test in the detection and the follow-up of skeletal metastatic lesions; it suggests the utility of the bone scan as a preliminary test, which allows a more rational use of the radiology.


Topography and clinical characteristics of malignant tumors of the urachus. Nine patients with urachal tumors have been reevaluated. There were 5 men and 4 women, whose ages ranged from 6 to 72 years. Surgery was performed in 4 cases, radiotherapy was used in 2 cases, and the remaining 3 cases were judged to be beyond therapeutic help. Three years after treatment, also all cases subjected to therapy were dead. The diagnostic delay, the inadequacy of treatment, and therefore the absolutely unfavorable prognosis of these tumors are discussed.


The value of radiotherapy in the treatment of vaginal carcinoma. Herein we report the retrospective survey of 48 consecutive unselected cases of vaginal carcinoma, mainly treated with radiotherapy in our Institute from 1959 to 1970. In this series irradiation was delivered almost always with a single and continuous application of sources of radium 226. Radiumtherapy treatment varied according to the extension in surface of the neoplasm, the clinical stage and especially the vaginal step involvement, considering the length of the organ. Despite the very good immediate response, failures of treatment locally or in paravaginal and pelvic areas were frequently observed, and success of the treatment after a brief follow-up was poor. The actuarial survival was 41.6% and 33.3% at 3 and 5 years, respectively. Stage I cases showed at the follow-up better therapeutic results (48.6% survival at 5 years) than stage II and III cases (28.3%). The poorest results were observed in neoplasms extended to the whole vagina, and all these patients died within 4 years of the beginning of treatment. This report stresses that radiotherapy of vaginal carcinomas demands individualization and a properly planned therapeutic program that combines external irradiation with renewed techniques of low dose rate and continuous irradiation with radioactive sources.


High resolution SEM of bacterial virus T7. High resolution "low-loss" scanning electron microscopy is a relatively new technique which permits an investigator to examine structures that were formerly visualized exclusively by transmission electron microscopy [1]. This paper presents some images of intact bacterial virus T7, viewed at the ultrastructural level. Due to the high resolution capibility of this technique, and the demanding physical prerequisites for visualization of the specimen, current specimen preparation techniques were modified in order to permit 1--2 nm resolution in surface mode. Using this method of microscopy, it is possible to view clearly this small bacteriophage (the smallest of the T-coliphages), adsorbed to its host bacterium, in a scanning mode at magnification (and resolution) comparable to TEM without resorting to the use of replicas, or reconstruction of a two-dimensional image.


Electron diffraction studies of the peptidoglycan of bacterial cell walls. Peptidoglycan of the gram-positive bacterium Lactobacillus plantarum was adsorbed onto hydrophilic crystalline films of graphitic oxide. Destruction by radiation damage was reduced by using a device which permitted scanning of the specimen for several hours through a focussed electron beam of low current density (2.5 X 10(-5) A/cm2). In additition to the sharp Debye-Scherrer rings caused by graphitic oxide, peptidoglycan causes a diffuse Debye-Scherrer ring in the region of 4.5 A. This can be interpreted as the packing periodicity of the peptidoglycan within the planes of the peptidoglycan sacculi.


The endocrine background of human renal cell carcinoma. IV. Glucocorticoid receptors as possible mediators of progestogen action. In order to investigate whether progestins may trigger tumour regression by a mechanism involving the glucocorticoid receptor, human renal cell carcinomas obtained from 15 patients were analysed for cytoplasmic glucocorticoid-binding components, using [3H] dexamethasone. The existence of glucocorticoid binders could be demonstrated in 10 out of 15 tumours studied. The average binding capacity was calculated and found to be 7.1 fmol/mg of cytosol protein. The apparent dissociation specificity experiments clearly cell-free system amounted to 1.9 X 10(-8) mol/l. The ligand specificity experiments clearly indicated that binding to these receptors is not restricted to glucocorticoids alone. Progesterone and aldosterone turned out to be moderate competitors for dexamethasone binding. Medroxyprogesterone acetate, the compound widely used in hormone therapy of advanced renal cancer in man, was demonstrated to be one of the strongest inhibitors of [3H] dexamethasone. It is concluded that binding of medroxyprogesterone acetate to glucocorticoid receptors might represent the primary mechanism of action of the compound in causing tumour regression.


Prognostic value of bone marrow acid phosphatase in prostatic carcinoma. The total and l-tartrate labile acid phosphatase were studied in 25 patients with carcinoma of prostate. The results were compared with the results from a control group. Serum acid phosphatase levels both in the control groups and in patients with prostatic cancer were lower than bone marrow acid phosphatase levels. This may be due to acid phosphatase released from blood cells during hemolysis. A positive correlation between serum and bone marrow acid phosphatase levels in patients with prostatic carcinoma was found. There was a significant rise in bone marrow acid phosphatase levels (above the normal range from the control group) in patients with advanced stage III and stage IV prostatic carcinoma with significantly increased serum levels. The levels of bone marrow acid phosphatase gave a supplementary diagnostic method in the diagnosis of prostatic carcinoma. A hypothesis that raised levels of bone marrow acid phosphatase are diagnostic of early metastasis from prostatic carcinoma is given.


Experimental embolization in kidneys: tissue concentration of antibiotics. In 7 dogs renal arterial embolization was performed using the Seldinger technique and an inflatable Swan Ganz balloon catheter. Methyl methacrylate and butyl-2-cyanoacrylate were injected. 6 months later, the tissue concentration of Cefazolin was measured in the embolized and contralateral kidneys and in serum 2 h after a short infusion of 1 h Cefazolin. The concentrations in serum and homogenized kidney tissue were determined by means of the agar gel diffusion method. Angiography showed no vessels in the embolized kidneys. Histologically there was total atrophy of the tissue, especially of the tubular system. The mean tissue level of Cefazolin in the embolized kidney was 70% of serum. The tissue concentration was always above 10 micrograms/g. This concentration is high enough to inhibit nearly all strains of E. coli, Klebsiella, Salmonella, Shigella and a large part of Proteus mirabilis, whereas nearly all gram-positive bacteria are inhibited at a lower concentration. The value and necessity of antibiotic therapy in embolized kidney cancer is discussed.


A new screening method for cystinuria. A simple and safe screening kit. The new kit (Urocystin) presented here utilizes the dark brown coloration which a neutral aqueous solution of cystine develops rapidly upon addition of nickel ion and sodium hydrosulfite (Na2S2O4). What is needed is pouring 4 ml of the urine sample into the kit, and the kit is able to judge without fail any urine sample with a cystine concentration of 50 microgram/ml or more as positive. No pretreatment of the sample is necessary. The kit contains no hazardous reagents at all. The kit has thus turned screening of cystinuria into an extremely simple affair.


Plasma levels of CEA as a prognostic marker in carcinoma of urinary bladder. Plasma carcinoembryonic antigen (CEA) levels were performed preoperatively by radioimmunoassay in 124 patients with histologically proved bladder carcinoma. The level of CEA was used to determine its prognostic value in patients with bladder cancer. The correlation of CEA levels with the stage of the disease, histology, and resectability was also studied. Values above 2.5 ng/ml were taken as abnormal. Active disease was associated with high CEA levels. All patients with CEA levels greater than 10 ng/ml died in less than 1 1/2 years, while all patients who survived 1 1/2-3 years had preoperative CEA levels less than 10 ng/ml. There was a prognostic significance for patients with transitional cell or squamous cell carcinoma. All patients with squamous cell carcinoma had CEA levels less than or equal to 10 ng/ml, and all patients with transitional carcinoma had preoperative CEA values greater than 10 ng/ml. A correlation between CEA levels and resectability of the primary tumor was found. This study indicates that, in bladder carcinoma patients, preoperative CEA levels greater than 10 ng/ml are of prognostic value, since all of these patients have died and all of the long-term survivors had levels of less than or equal to 10 ng/ml.


Chronic interstitial nephritis as a cause of tumours of the upper urinary tract. A hypothesis. Urothelial tumours of the upper urinary tract are rare, but their incidence increases in subjects suffering from phenacetin nephropathy, Balkan nephropathy and infected pyelic calculi. In order to act on the urothelium through the urinary way, a cancerigenic substance must (1) be eliminated with the urine; (2) be absorbed by the urothelium; (3) remain in the tissue for a certain length of time. Any situation leading to a prolongation of intra-tissue stasis favours the action of carcerigenic substances. The hypothesis is put forward that interstitial nephritis, by impeding the reabsorption from the pelvis by pyelo-lymphatic reflux and by making more difficult the lymphatic drainage of the pyelic and ureter walls, leads to a condition favouring carcinogensis, especially if the subject is exposed to contact with even not very active carcinogens. It is possible to foresee that in future the incidence of upper urinary tract tumours should increase in chronic uraemic patients under dialytic treatment and in transplant patients.