Edward C. Hughes

Organ culture of human endometrium: Effect of ovarian steroids☆☆☆

Abstract Glycogen turnover was determined in 86 normal and 52 sterility patients through the chemical analysis of endometrial glycogen content, glycogen synthetase and glycogen phosphorylase activities. Specimens from 38 of these normal patients as well as 31 sterility patients were also subjected to the organ culture process. These studies suggest to us that (1) an organ culture system is an ideal avenue of approach for the study of this glandular epithelial tissue and (2) that estradiol is not solely responsible for the increase in glycogen deposition in human endometrium, but that progesterone following estrogen priming is essential for a complete response of glycogenesis. This study further demonstrates in organ culture that estradiol-induced proliferative changes in the tissue, while progesterone not only caused cellular alterations but also induced an increase in glycogen deposition. Preliminary studies with the enzyme systems in organ culture suggest a role of progesterone in phosphorylase activity, particularly during the secretory phase where a controlled breakdown of glycogen is desired.

Carbohydrate pathways of the endometrium: Effects on ovular growth

Abstract From these data, it can be said that the metabolism of carbohydrates and proteins is deranged in patients with the history of sterility and repeated abortions or congenital malformations. In these cases, all other conditions which could account for sterility or abortion were found to be in order. The abnormal histochemical changes, the alterations in the Krebs citric acid cycle, the decreased secretion of glucose-6-phosphatase and alkaline phosphatase together with the inability of the tissue to synthesize amino acids to RNA and DNA and hence on to the various enzymes needed in the carbohydrate metabolism probably indicate that the endometrial tissue in these patients was not carrying on the process of glycolysis or glycogenesis in a satisfactory manner. This may be due to (1) lack of estrogen and progesterone due to pituitary or ovarian failure, (2) anatomic, structural, and physiologic abnormality of the uterus and/or endometrium, (3) poor blood supply due to capillary disease, (4) a combination of all three factors. Any one of these factors may affect the amount of glucose and oxygen delivered to this tissue. In some instances these abnormal changes can be corrected by the proper use of the ovarian steroids.

Chromosome constitution of human endometrium

Abstract Samples of endometrium from 21 normal women, 21 patients with a history of sterility, 21 women with problems of repeated abortion, and 3 patients ∗ ∗Since statistical analysis was completed, endometrium from 5 more patients with functional uterine bleeding have been studied and similar chromosome counts were obtained. with functional uterine bleeding were grown in tissue culture. Samples of the cervices of 9 of these patients were also grown in vitro by similar methods and used as controls. From these cultures, 2,482 metaphases were obtained and chromosomes were counted. Karyotypes were studied in the technically satisfactory ones. Aneuploidy was present in all types of endometrium with the largest percentage of cells with abnormal chromosome numbers being of the hypodiploid type. There was also a statistical difference in the proportion of cells with 46 chromosomes between normal endometrium and cervix, between the normal versus the sterility and abortion groups with the greatest difference existing between the bleeders and normal, sterility, and abortion groups. The karyotypes of cells with 46 chromosomes were normal, while there was a random loss of chromosomes in the hypodiploid group. The aneuploidy present in all types of endometrium is apparently the physiologic characteristic of the tissue and the regular sloughing of the endometrium may be necessary to avoid pathologic states which create excessive bleeding and may lead to malignant change in the endometrium.

Some recent observations concerning the toxemias of pregnancy.

Abstract 1. 1. There is no increase in antidiuretic activity of serum during normal and toxemic pregnancy. 2. 2. “Total corticosteroid” increases throughout pregnancy, reaching a peak before delivery. Freely water-soluble corticosteroid increases throughout pregnancy. A poorly water-soluble material appears during the first trimester and increases until the end of pregnancy, constituting 25 per cent of the corticosteroid. 3. 3. In toxemia, corticosteroid is considerably increased above the level found in normal pregnant women at the same stage of pregnancy. The poorly water-soluble component is also considerably increased above that seen in the normal, respresenting approximately 35 per cent of the total corticosteroid. In one patient with eclampsia, paper chromatography demonstrated a material which has a rate of flow characteristic of 3-oxygen-containing steroids. 4. 4. The possibility that this material might play a part in the etiology of eclampsia is discussed. 5. 5. The placental steroid increases during pregnancy, while the levels of chorionic gonadotrophin remain at a low level. 6. 6. These steroids together with the adrenal steroids may affect the general metabolism of the body and also may sensitize the kidneys so that added factors, perhaps from the placenta, may extend the lesions in these organs so that the function of the kidneys is temporarily altered. Prolonged action of all factors may permanently damage the kidneys. 7. 7. Reversed excretion of placental hormones during the latter months of pregnancy is noted in toxemia, and may reflect failing placental physiology. 8. 8. These studies have significant clinical application and direct our attention to the fact that sodium chloride, particularly, should be restricted as early in the gestation as the one hundred twentieth day.

Effect of oral contraceptives on human endometrium in culture

Abstract Addition of oral contraceptives to the medium of human endometrial explants in organ culture caused histologically, histochemically, and biochemically detectable changes in the tissue. These alterations were similar to the response obtained in vivo and indicate that the synthetic hormones have a direct effect on the endometrium in addition to any other mode of action. The effects of the contraceptives in organ culture included proliferative and secretory changes similar to those cause by the natural ovarian hormones, increased deposition of glycogen in the glandular epithelium and in the stromal cells, and histologic changes in the stroma around the glands and the blood vessels.

Diminished plasma 17-ketosteroid concentration in pregnancy.

Summary 1. Plasma neutral 17-ketoste-roid determinations were made on control group of 33 normal men and women, and on experimental group of 30 pregnant women at term. 2. Adult controls showed mean plasma value of 61 μg/100 ml (S.E. = 5). No significant difference was found between values obtained on normal men vs. normal, nonpregnant women (P >.05). 3. Pregnant women showed a mean plasma value of 18 μg/ 100 ml (S.E. = 2). This value is significantly lower than the mean of control adults (P <.001). 4. From the results of plasma analyses it is concluded that there is a physiological reduction in maternal elaboration of 17-ketosteroids during the last 24 hours of pregnancy. The data of other workers on urinary excretion of 17-ketosteroids in pregnancy (1-4) suggest that this finding may hold for the last third of pregnancy. Renal clearance studies will be necessary before final conclusions can be drawn. 5. It is postulated that the inner cortex of the fetal adrenal and/or the placenta take over production of 17-ketosteroids during the latter part of pregnancy.

Effect of experimentally induced cervical cancer upon the nucleic acid and carbohydrate metabolism in various organs of mice

Abstract Squamous cell cancer was induced in the uterine cervix of S-3H strain of mice within 8 to 12 weeks after implantation. The experiments compared the metabolism of carbohydrates and nucleic acids in the cervical cancer with normal cervical tissue. It determined the effect of cancer growth upon the carbohydrate and nucleic acid metabolism of other essential organs by measuring the following enzymes and metabolites. Lactic acid content, glycogen, glycogen synthetase, glucose-6-phosphate dehydrogenase, and TPN-dependent isocitric dehydrogenase. These substances were significantly altered in the spleen, kidney, and cervical cancer of the cancer animals. The relative rate of mitosis in all structures was evaluated by assaying the tritiated thymidine at various time intervals after intraperitoneal injection. The DNA and RNA content was estimated in all by the quantitative extraction of the nucleic acid fraction. Marked changes in the metabolic rate and nucleic acid metabolism was noted in the spleen, kidney, lymph nodes, and cervical cancer.

Diabetes and Pregnancy

The incidence of diabetes increases with age and with parity. Since the disease alters most of the endocrine and metabolic changes of normal pregnancy, complete prenatal care with rigid control of the diabetes and timely delivery are most important in managing the diabetic pregnant patient. It is considered to be good policy to give the infant glucose for at least the first four or five days.