Fluorescent in situ hybridization (FISH) on paraffin-embedded placental tissues as an adjunct for understanding the etiology of early spontaneous abortion.

OBJECTIVES: An investigation of first-trimester spontaneous abortions (SAs) for those cases in which karyotype is not available was designed to test the efficiency of fluorescence in situ hybridization (FISH) on paraffin-embedded tissues combined with pathological examination for understanding the etiology of SAs. METHODS: Pathological examination of 202 placental tissues from SAs was performed. FISH analysis was then carried out on paraffin-embedded tissue sections from the same abortion products with probes specific for chromosomes 13, 16, 18, 21, X, Y. RESULTS: FISH could be achieved in 196 cases (97%). After pathological analysis alone, the etiology of SAs was evoked in 40 cases. The suspected diagnosis was confirmed by FISH in 26 cases (13.2%). After combined pathological and FISH analysis, the etiology of SAs was identified in 83 from the 196 cases (42.3%) with the probe set used. CONCLUSION: The present study demonstrates the value of FISH on paraffin-embedded tissues as an adjunct for understanding the etiology of SAs for those cases in which karyotype is not available. Combination of pathological and FISH analysis increases the yield of diagnosis by a factor of 3.2. The results also demonstrate that predictions of the karyotype from pathological examination should be avoided.

Diamine oxidase activity and biochemical markers in human seminal plasma.

Diamine oxidase (DAO), an enzyme which degrades polyamines, is present at a very high level in human seminal plasma and is assumed to come mainly from the prostate. The possible relationships between DAO activity and biochemical markers of accessory sex glands were evaluated in 139 men in barren marriages. Four groups were formed: normozoospermic (n = 41), asthenozoospermic (n = 29), oligoasthenozoospermic (n = 35) and azoospermic (n = 34). DAO activity was the highest in the asthenozoospermic group and was significantly different from that in the azoospermic one. For all specimens, a positive correlation was demonstrated between DAO activity and the prostatic markers citric acid and acid phosphatase. However, DAO activity was correlated with citric acid only in the oligoasthenozoospermic and the azoospermic groups. Acid phosphatase and citric acid were linked in all groups. These results implicate the DAO enzyme in changes in sperm metabolism leading to a loss of motility and suggest that DAO comes partly from the upper part of the genital tract (testis and/or epididymis), in addition to the prostatic gland secretion, accounting for the absence of correlation with prostatic markers in normozoospermic and asthenozoospermic groups.

1H nuclear magnetic resonance of human seminal plasma in in vitro fertilization attempts: use of automatic spectrum analysis.

Seminal plasma affects the quality of spermatozoa. The possible use of 1H nuclear magnetic resonance (NMR) analysis of human seminal plasma to determine a full biochemical profile that would allow the prediction of successful in vitro fertilization was examined. Two seminal plasma patterns (GI and GII) were found. The main compounds in GI were glycerophosphorylcholine, citrate and lactate, and in GII, choline and amino acids. These patterns did not fit into the usual spermogram classes. However, GI exhibited the higher pregnancy rate, suggesting that the equilibrium established between the components in seminal plasma affected not only the quality of spermatozoa but also embryonic quality.

Diagnostic value of erythrocyte-free polyamines and histaminemia in malignant hepatic tumors and in liver cirrhosis.

The present study tries to evaluate the diagnosit value in malignant hepatic tumors of polyamines, of which the relationship with cellular kinetics is known, and histamine, of which catabolism follows a similar pathway. One hundred and fifty six patients were studied: 53 with malignant liver tumors (27 primary, 26 metastatic) and 103 with non-tumoral liver diseases of which 65 were cirrhotic and 38 non-cirrhotic. Erythrocyte polyamines (spermidine and spermine) and histamine levels were assayed. The results indicate the following. 1. Polyamine levels were significantly increased (a) in cirrhotic patients, not only when compared with controls (p less than 10(-8)), but also when compared with the non-cirrhotic patients (p less than 10(-7)); (b) in primary malignant hepatic tumors (p less than 10(-3)). 2. Histamine was significantly increased (a) in the non-tumoral liver diseases (p less than 10(-4)), but with no difference between cirrhotic and non-cirrhotic patients; (b) in the secondary malignant tumor patients, histamine levels were lower than in primary tumor patients (p less than 0.04). 3. There was no correlation, in all groups studied, between polyamine and histamine levels. These results suggest the following practical implications. 1. For non-tumor liver diseases, appreciably increased polyamine levels may represent a further argument favoring a cirrhotic condition. 2. In diagnosing hepatic scintigraphic defects, increased polyamine levels would suggest a primary malignant hepatic tumor; low histamine levels are more in favor of a secondary malignant hepatic process.

Red blood cell polyamines in mice bearing the Lewis lung carcinoma (3LL) and in patients with bronchopulmonary cancers.

Experimentally, during Lewis lung carcinoma (3LL) growth, the red blood cell (RBC) polyamine levels increase with tumor volume and are inversely correlated to tumoral concentrations of spermidine and spermine. The RBC level of spermidine is continually correlated to both the volume and the tumoral concentration of this polyamine. Clinically, high levels of RBC polyamine are observed in cases of squamous-cell carcinoma or anaplastic cancer, and not in cases of adenocarcinoma. RBC polyamine levels only permit the establishment of statistical differences between groups of patients. Therefore, clinical use of these molecules as tumor markers depends on an understanding of polyamine distribution within blood.

In vitro studies on the entry of polyamines into normal red blood cells.

Polyamines are mainly transported in the blood by erythrocytes: Putrescine, spermidine and spermine can be taken up in vitro by red blood cells (RBC); their entry is greater in the presence of serum than in the presence of plasma, and spermine entry is lower than that observed for the two other polyamines. In the presence of serum, the affinity of RBC for spermidine is 30 fold greater than that for putrescine. The majority of RBC polyamines are present in the hemolysate and are not complexed to high molecular weight material. At + 4 degrees C the polyamine uptake is considerably reduced and for putrescine and spermine practically non existent, but it seems that it is internalization rather than binding which constitutes the dependent step. Though intracellular spermidine and spermine levels reflect differences in uptake rather than in outward flux across the cell membrane, the values of putrescine appear to be the resultant of influx and efflux. The presence of specific receptor sites for polyamines visualized by SEM on the surface of RBC using latex-putrescine spheres, confirms the results obtained with labelled polyamines. Therefore, only the understanding of the polyamine repartition inside the blood compartments would permit the clinical use of those molecules as non statistical tumor markers.

Polyamines in human brain tumors. A correlative study between tumor, cerebrospinal fluid and red blood cell free polyamine levels.

Polyamine levels have been studied in brain tumor patients. We focused our study on the relationship between tumor, cerebrospinal fluid (CSF) and red blood cell (RBC) polyamine levels. Our results are the following: Polyamine levels in CSF are consistently increased, whatever the histological type may be. The highest tumoral concentrations are found in medulloblastoma. In glioblastomas, the RBC spermidine levels are higher than in the other types of tumors and there is a highly significant correlation between the spermidine/spermine ratio in tumor and RBC. Therefore, RBC polyamine determination might be of clinical interest in the monitoring of patients with glioblastomas.

[Inhibin, a second testicular hormone (author's transl)]. / L'inhibine, seconde hormone testiculaire.

Germinal epithelium secretes a peptide hormone, inhibin, that regulates FSH secretion. This substance originates inside the seminiferous tubules, from the Sertoli cells, and is transported towards the head of the epididymis where it is reabsorbed, and thus passes into the bloodstream. Inhibin blocks FSH pituitary secretion; its effect on the hypothalamus is debatable. Inhibin also exists in the female, in ovarian follicles and may have a role in follicular growth regulation.

Role of the epididymis in reabsorption of inhibin in the rat.

The effects of efferent duct ligation, orchidectomy and orchido-epididymectomy were compared at 36 h. This showed that the rat epididymis is capable of maintaining a negative feedback control of FSH secretion. Epididymal extracts contained a substance of tubular origin capable of reducing FSH secretion. This substance is a protein and is found in the caput region but not in the caudal region of the epididymis. A new role can be assigned to the head of the epididymis: that of reabsorbing, together with testis fluid, the inhibin produced by the seminiferous tubules, which thus passes into the bloodstream and provides a feedback regulation of plasma FSH.

[Synergistic effects of ethynylestradiol and norethisterone on the formation and lengthening of uterine lace in immature rabbits]. / Effets synergiques de l'éthynylestradiol de la noréthistérone sur la réalisation et la prolongation de la dentelle utérine chez la lapine impubère.

We have shown, through a test proceeding from the McPhail test, in the immature rabbit female, the existence of succession and simultaneity synergisms between ethynylestradiol and pure or impure norethisterone, given by oral route. The estrogen, administrated before the progestative, is able to potentialize the action of the latter on the building of the endometrial lace in 5 days, more than on the lengthening of it in 15 days. Ethynylestradiol, absorbed with norethisterone, strengthens the effects of the latter on the second phenomenon more than on the first one. Absolute and relative doses of the estrogen and of the progestative differ only lightly according as norethisterone is pure or not pure, because of the more potent estrogenicity of the impure norethisterone.

[Testing synergism between ethinyl estradiol and norethisterone on the histamine-induced deciduoma and vaginal mucification of castrated rats]. / Essais de synergie entre l'éthynylestradiol et la noréthistérone sur le déciduome histaminique et sur la mucification vaginale de la ratte castrée.

Through the histaminic deciduoma test and a vaginal mucification test, we have endeavoured to show that, in spayed female rats there exists a possibility of synergism between ethynylestradiol and pure norethisterone per os. No synergism could be shown in the former test, owing to, or so it seems, a lack of deciduogenic power proper to norethisterone. The latter test showed, indeed, that ethynylestradiol can, in low quantities, bring about a synergism on the mucifying properties of norethisterone, whereas, in large quantities, it creates an antagonism on the aforesaid properties.

[Effects of ethinyl estradiol compared with norethisterone (pure or contaminated) on vaginal keratinization of castrated rats]. / Actions comparées de l'éthinylestradiol et de la noréthistérone pure ou non sur la kératinisation vaginale de la ratte castrée

Norethisterone samples are often contaminated by estrogen traces which can be measured by mass spectrometry and which invest the progestative with estrogenic properties in the Allen and Doisy test. Therefore, one had better, in pharmacodynamic experimenting, use samples that have been purified by chromatography or that are pure out of synthesis. However, one will not forget these are not without some estrogenicity, a fact which seems linked to the very nature of the norethisterone molecule.

PIP: Compared were norethisterone from Roussel to norethisterone purified by silicic acid chromatography, containing less than .025 percent mestranol, to norethisterone from Syntex synthesized by a procedure that would not lead to contamination with mestranol in an adaptation of the Allen-Doisy rat vaginal cornification test. Castrated Wistar-Commentry rats were given the steroids orally in sesame oil for 3 days and vaginal smears were checked on the 4th day. Smears were considered positive if they contained nucleated or anucleated epithelial cells; negative if they had any polynuclear cells. Dose-effect studies showed that Roussel norethisterone was 1/200 as estrogenic as ethinyl estradiol. The purified norethisterone and the mestranol-free norethisterone were 1/1000 as estrogenic as ethinyl estradiol. Thus the estrogenic effects of norethisterone are probably due to mestranol contamination, rather than to bioconversion within the organism, as has often been claimed.