Nagoya Journal of Medical Science

This report deals with the relationship between liver L-Alanine: 2-oxoglutarate aminotransferaes (EC 2. 6. 1. 2) activity and RES function.
In fasted rats, decrease in Δ⁴-3-ketosteroid hydrogenase (EC 1. 3. 1. 3 and 1. 3. 1. 4) and phagocytic activity and increase in alanine aminotransferase activity were noted. On the other hand, increase in Δ⁴-3-ketosteroid hydrogenase and phagocytic activity and decrease in alanine aminotransferase activity were observed in RES stimulated ones. Possible mechanism for these changes in alanine aminotransferase activity was discussed in relation to the role of reti- culoendothelial system in steroid hormone metabolism.

Based on histological investigation of the testes in 270 patients with functional infertility, in 13 patients with stenotic lesion and in 11 men with proven fertility, the following conclusions were obtained.
1) Testicular biopsy should be of first choice in the examination of male infertility. Epididymo-vaso-vesiculography to prove the patency of the seminal tracts is not always necessary. It should be done only on patients with testicular histology which can be expected the efficacy of treatment.
2) The size of biopsied specimen of the testis should be larger than half a pea, and open biopsy should be made in order to obtain a satisfactory size of specimen.
3) Bouin's solution is best for fixation of testicular tissue, and Carnoy's solution might be used, if Bouin's solution is not available. Zenker's solution and formol are not satisfactory for this purpose.
4) Biopsy is not necessary for both testes, and unilateral biopsy is sufficient to reveal the histological character of both testes, if no difference exists between the both testes by scrotal palpation.
5) In patients with sperm concentration much higher than 20 x 106 per ml, information obtained by testicular biopsy is of little value.
6) In patients with testis smaller than thumb-head size, testicular biopsy shows severe damage, which discourages treatment.
7) Testicular injury due to biopsy procedure may lower the sperm concentration in patients with very low sperm counts. The effect may be irreversible.

Histological investigation of the testes of ~14 patients with azoospermia, 56 patients with oligozoospermia and 11 men with known fertility was made. In 24 of these cases, 17-ketosteroids and pituitary gonadotropin in urine were determined. Based upon the results of these studies, pathological problems of functional infertility in human male were discussed.
1) No qualitative difference in the histological findings of testes was observed between azoospermia and oligozoospermia.
2) All atrophic testes, smaller than thumb-head by scrotal palpation, showed highly damaged spermatogenesis.
3) In 27% of the patients with oligozoospermia, with less than 20x10⁶ per ml in sperm concentration, no spermatozoa were found in the seminiferous tubules of the biopsied specimen. The evidence shows that possibility of histological misdiagnosis is inevitable, . though the artefact due to technical procedure has to be taken into account.
4) In 19% of non-obstructive azoospermia, active spermatogenesis was recognized in the majority of the seminiferous tubules. The epididymo-vasal secretion of them did not contain any spermatozoa. The evidence calls for the necessity of examination of the rete testis and the distal end of the seminiferous tubules.
5) In normals, the mean thickness of the tubular walls ranged from 4.5 to 5.5 µ. In hypofunctional testes, the distribution could be grossly divided into two groups, one group with the tubular walls as thin as the normals and the other the group with tubular walls thicker than 6.5 µ.
6) In normals; the mean diameter of the seminiferous tubules ranged closely around 150 µ. In oligozoospermia, the distribution was grossly divided into two groups, one with the tubular lumina as wide as the normals and the other with narrow tubular lumina, less than 110 µ in diameter.
7) Generally, the development of germinal cells was well maintained in tubules with thin walls and wide lumina, and Leydig cells were distributed normally around these tubules. On the contrary, germinal maturation was lowered in tubules with fibrously thickened walls and narrow lumina, and Leydig cells were hyperplastic to various degrees.
8) The histological picture of testis in infertile human male closely resembles to experimentally allergic lesion of testis in guinea-pig or rat.

Mast cells appear to play significant roles in physiological and pathological conditions. Some functions of mast cells may relate to liver cirrhosis, which is characterized by degeneration, necrosis and regeneration of parenchyma as well as fibrosis, but little information regarding this association has been obtained. This study was carried (Jut in order to clarify the pathogenesis of liver cirrhosis as related to the distribution and the role of mast cells. Fifty-five cases of liver cirrhosis were divided into five groups, according to Kim's morphological classification and the mean values with the standard errors in each group were deter- mined with regard to the number of mast cells, the relative volume of the trabecula and the liver weight. A statistical analysis of these determinations was also performed. The results of the present investigation revealed that a greater number of mast cells were found in the I-B and the n-B groups in which a more marked regeneration was histopathologically recognized, and that in these groups a statistically significant correlation existed between the number of mast cells and the liver weight which mostly increased with regeneration. These findings strongly suggested that mast cells in human liver cirrhosis had a close relation- ship to regeneration of hepatic parenchyma followed by occasional incidence of malignant hepatomas.

Several drugs were infused intravenously into rabbits to produce diuresis or antidiuresis, and the concentration of sodium and potassium was determined in the cortex, medulla and papilla of the kidney.
The papillary sodium concentration and the cortical potassium concentration were decreased in the furosemide-infused group, while the sodium content was decreased only in the papilla in the ethacrynic acid-infused group. These agents have no effect on the sodium content in the cortex in spite of producing marked diuresis.
The medullary and papillary sodium concentration and the medullary potassium concentration were decreased in the 20% mannitol-infused group. In the 50% glucose-infused group, the sodium content was reduced in the cortex, medulla, papilla, cortical mitochondrial fraction and microsomal fraction, whereas the potassium content was depressed in the cortex, medulla and cortical microsomal fractions. Changes in the sodium and potassium concentration were observed over wider range as compared with the furosemide and ethacrynic acid-infused groups. The above change could be due to the difference in the concentration of infusates.
The medullary potassium concentration was raised, but no change was observed in the sodium concentration in the group given low doses of vasopressin (0.1 U/kg ). The infusion of a high dose of vasopressin (10 U/kg ) caused an increase in the sodium concentration in the cortex, cortical mitochondrial and microsomal fractions, while even lower dose of vasopressin produced an increase in the medullary potassium. The above result would suggest that vasopressin might act on the reabsorption of potassium stronger than that of sodium. Judging from the fact that the renal blood flow would be decreased by vasopressin, the energy required to reabsorb sodium and potassium may be partly supplied from other sources than ATP, as reported in Part II of this study.

In Part I the concentrations of sodium and potassium were examined under the same conditions of diuresis and antidiuresis. In the present paper Na-K ATPase activity was assayed under these conditions in order to evaluate the transport of sodium and potassium by the renal tubules.
Both furosemide and ethacrynic acid decreased Na-K ATPase activity in the whole homogenates of the cortex, medulla and papilla, in the cortical mitochondrial and microsomal fractions, by intravenous administration. This finding indicates that these agents produce marked diuresis by inhibiting Na·K ATPase activity.
With respect to osmotic diuresis both 5% and 20% mannitol also decreased Na·K ATPase activity in all of the parts. On the other hand, both 5% and 50% glucose did not affect Na-K ATPase activity. Judging from these results, it would be sure that mannitol produces diuresis by inhibiting· Na-K ATPase and has a different mode of action on diuresis from that of glucose.
With respect to antidiuresis, a low dose of vasopressin (0.1 U/kg) did not affect Na-K A TPase activity, but a high dose of vasopressin (10 U/ kg) decreased it in the whole homogenates of cortex, medulla and papilla, in the cortical mitochondrial and microsomal fractions. This decrease would be partly dependent upon the renal blood flow. It was reported in Part I that the sodium and potassium contents were increased, but Na-K ATPase activity was decreased in this work. This result would show that Na-K ATPase is independent of the reabsorption of sodium and potassium caused by administration of vasopressin. It would also be supposed that the energy required for reabsorption of sodium would be supplied at least in part by a pathway other than of ATP.

Aminoglutethimide (Elipten, Ciba,) interferes with the biosynthesis of adrenocortical hormones by blocking the conversion of cholesterol to pregnenolone. Male albino rats were fed with the diet containing aminoglutethimide and were killed after various time intervals ranging from 24 hours to 7 days. The adrenal cortices of these animals were studied both by light and electron microscopy. Special effort was made to determine the intracellular localization of free cholesterol by an electron microscopic histochemical method using the digitonin reaction.
The animals killed 24 hours after the initiation of the feeding were found to have small myelin figures in contact with the mitochondrial membrane in fasciculata cells which might reflect compensatory rise of ACTH. The animals fed with the diet for 4 to 7 days were shown to have enlarged adrenal cortex. Light microscopic examination revealed an increase of the lipid globules both in size and number in the zona fasciculata. Ultrastructural studies showed swollen, hypertrophied, bizarre prophiles of the mitochondria with the formation of the vacuoles. The electron microscopic histochemical method demonstrated digitonincholesterol crystals localized in the vacuoles of the mitochondria, in the perimitochondrial areas and in the lipid globules.
These findings correlate well with the biochemical data which indicate the intracellular site of steroid bicsynthesis and lend morphological support to the view that the biosynthesis of pregnenolone from cholesterol takes place within the mitochondria. These results indicate that aminoglutethimide causes accumulation of cholesterol in the vacuoles of the mitochondria.

The following experiments were made to examine the increased permeability of vasa vasorum. Dogs of approximately 10 kg body weight were anesthetized by intraperitoneal injection of 30 mg/kg of pentobarbiturate. One ml of 5 µg/ml bradykinin solution in saline was iufiltrated around the tissues of the exposed carotid arteries and iliac arteries respectively. Immediately afterwards 50 ml of homologous serum protein combined with fluorescent· dye was injected intra· venously. The vessels were removed 2 to 8 minutes after the injection of the dye. The removed vessels were observed under a fluorescence microscope. The intima was peeled to make the material transparent which was essential for the microscopic observation. Control experiments were made by using normal saline in the same operative procedure. Vasa vasorum of the arteries were distinctly seen emitting green fluorescence. Leakage was most prominent from venules of 20 to 30 µ in diameter, especially at the branching junctions, but no leakage was found along arterioles. The fact that bradykinin increases permeability of vasa vasorum suggests an important role on the venular side of vasa vasorum in the pathogenesis of vasular diseases.