H Sobis

Angiogenic activity of peritoneal fluid from women with endometriosis.

OBJECTIVE To investigate the presence of angiogenic factors in peritoneal fluid (PF) and follicular fluid (FF). DESIGN The PF samples of 48 women with (n = 24) or without (n = 24) endometriosis were investigated. Angiogenesis was assayed using the chorioallantoic membrane of 11-day-old fertilized chicken embryos. Glass fiber prefilters impregnated with the fluids were placed on the chorioallantoic membrane. MAIN OUTCOME MEASURE The vascular reaction was analyzed by an independent observer after 72 hours. RESULTS There was a positive reaction in 58.3% of the PF from women with endometriosis and in 12.5% of the women without endometriosis. No correlation was found between the angiogenic response and the severity of endometriosis. The reaction remained after charcoal treatment of the PF. Positive reaction was found in three of six FF samples. CONCLUSION The PF of women with endometriosis contain more angiogenic factors than PF from women without endometriosis. This angiogenic activity could be important for the further outgrowth and progression of the lesions.

Efficacy of (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)-cytosine and 9-(1,3-dihydroxy-2-propoxymethyl)-guanine in the treatment of intracerebral murine cytomegalovirus infections in immunocompetent and immunodeficient mice

The efficacy of HPMPC and DHPG against systemic and intracerebral murine cytomegalovirus (MCMV) infections was examined in immunocompetent NMRI mice and in mice with severe combined immunodeficiency (SCID). HPMPC proved to be far superior to DHPG in preventing mortality and growth retardation in MCMV-infected NMRI mice even when given as a single dose on the day of infection. In intraperitoneally infected SCID mice, HPMPC administered as a single dose of 2, 10, 20 or 50 mg/kg per week increased the survival period of the mice by 22, 49, 77 and 156 days, respectively. In contrast, DHPG at daily doses of 10, 20 or 50 mg/kg for five consecutive days every week did not delay death by more than 13, 17 and 21 days, respectively. About one week before the MCMV-infected SCID mice (treated with either DHPG or HPMPC) died, they developed signs of neurological disease and intranuclear inclusion-bearing cells were found in their brains. The virus that was recovered from the brains of these mice did not prove to be resistant to HPMPC or DHPG. Only the virus recovered from the brains of mice treated with HPMPC at a dosage of 50 mg/kg/week had a slightly decreased susceptibility to HPMPC. When HPMPC (50 mg/kg for 4 consecutive days) was administered to SCID mice at the time when neurological symptoms became apparent, death of the animals could be delayed by another 35 to 40 days.

Inhibitory effect of a broad-spectrum antiviral agent, (S)-9-(2,3-dihydroxypropyl)adenine, on spermatogenesis in mice

Abstract In subacute toxicity studies in NMRI mice, the broad-spectrum antiviral agent (S)-DHPA [(S-9-(2,3-dihydroxypropy)adenine] did not exert any specific organ toxicity, even when administered at doses up to 1 mg/g body wt, except for testicular germinal aplasia. This effect was obviously accompanied by sterility and could be established in both young (3-week-old) and adult (6-week-old) mice which had received the drug (at 1 mg/g/day) in their drinking water for a period of 21 days. Under these conditions, the testicular dysfunction generated by (S)-DHPA was apparently fully reversible: fertility was restored within 5–6 weeks after the end of drug treatment, and the resulting offspring did not show an increased death or malformation rate as compared to the offspring of untreated males. (S)-DHPA did not affect the fertility of female mice and its deleterious effects on male fertility were only observed at a dosage of 1 mg/g, and not at a dosage of 0.3 mg/g or lower.

Effect of human and mouse interferon and of polyriboinosinic acid·polyribocytidylic acid on the growth of human fibrosarcoma and melanoma tumors in nude mice☆

Abstract Human leukocyte interferon, human fibroblast interferon, mouse interferon and an interferon inducer, polyriboinosinic acid·polyribocytidylic acid (poly(I)·poly(C)), were examined for their effect on the growth of two human tumor cell lines in athymic nude mice. The human tumor cell lines were derived from a fibrosarcoma (designated HT- 1080 ) and a nonmelanotic melanoma (designated A- 375 ). Repeated doses of 5 mg/kg of poly(I)·poly(C), administered every other day starting 1 day after tumor cell inoculation, caused a significant inhibition of tumor growth. On the contrary, human (leukocyte or fibroblast) interferon and mouse interferon, at doses up to 5 × 10 6 units/kg , did not affect the growth of either of the two human tumor cell lines tested.

Teratoma induction in mice and rats in relation to the age of the visceral yolk sac.

Abstract The morphological and biological characteristics of yolk sac-derived mouse teratomas are compared to those induced in rats. In both species, similar well differentiated adult tissues were recorded. The differential potential of the yolk sac was identical between the different mouse and rat strains tested but was very much dependent upon the age of the yolk sac at the time of operation. Killing of the primary germ cells by Busulphan treatment did not influence the development of these teratomas.

Histochemical differences in expression of X-linked glucose-6-phosphate dehydrogenase between ectoderm- and endoderm-derived embryonic and extra-embryonic tissues.

We examined the activity of X-linked glucose-6-phosphate dehydrogenase (G6PD) in concepti of the enzyme-deficient mutant and wild-type C3H mice. By using different crosses between the G6PD-deficient homozygous, heterozygous, or wild-type females and hemizygous or wild-type males, we confirmed the inactivation of one of the two X chromosomes in female concepti by a histochemical method. With this technique, a dual (G6PD + or -) cell population could be observed in the tissue sections. We demonstrate that the paternal X chromosome is inactivated in the endoderm of parietal and visceral yolk sac and in the trophoblast, whereas in the embryo and in the yolk sac mesoderm this inactivation is random. Our results confirm biochemical observations showing that only the maternal X chromosome is expressed in all derivatives of trophectoderm and primitive endoderm, whereas derivatives of the primitive ectoderm show random X chromosome expression.

A Rat Cytotrophoblast Antigen Defined by a Monoclonal Antibody

ABSTRACT: A monoclonal antibody that recognizes specifically a cytotrophoblast antigen was obtained. The monoclonal antibody 22H6 was tested on rat choriocarcinoma (in vivo, in vitro), normal placenta, ectoplacental cone, blastocysts, and several normal organs. The antigen was detected on frozen sections and on tissue culture by indirect immunofluorescence. The monoclonal antibody 22H6 reacts with the cytotrophoblasts of rat choriocarcinoma. The giant cells do not display a positive reaction. It is not expressed on other tumors than choriocarcinoma. In adult rats the only cells revealing a positive reaction are the hepatocytes and the epithelial cells lining the small intestine. In the pregnant rat, the antigen is expressed on the cytotrophoblasts of the junctional zone in the placenta, but not on the giant cells. The mab reacts only with the small trophoblast cells of the ectoplacental cone, but not with trophectoderm of blastocyst. The mab has an IgG2b isotype and is not cytotoxic for choriocarcinoma cells in a complement‐dependent cytotoxicity test. The described monoclonal antibody is to our knowledge the only known marker of rat benign and malignant cytotrophoblast.

Normal and Malignant Trophoblasts Do Not Recruit Granulated Metrial Gland Cells

A possible relationship between the development of granulated metrial gland (GMG) cells and trophoblast was studied. Trophoblast implanted in ectopic sites (e.g. kidney capsule) did not induce decidua and did not recruit GMG cells. Only when injected in utero did trophoblast lead to the development of decidua and to the recruitment of GMG cells. With malignant trophoblast (choriocarcinoma cells) similar results were obtained as with normal trophoblast both after ectopic or after in utero injection. The presence of decidua, but not the development of a conceptus or the outgrowth of trophoblast, seems to be required for the differentiation of GMG cells.

Influence of Different Growth Factors on a Rat Choriocarcinoma Cell Line

The influence of epidermal growth factor, insulin-like growth factors I and II, insulin, transforming growth factor beta 1 and transferrin on the growth of a postgestational rat choriocarcinoma was examined by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. The cell line was cultured in RPMI 1640 medium supplemented with fetal calf serum, beta-mercaptoethanol, glucose, sodium pyruvate and antibiotics. The experiments were done in media supplemented with 10% (optimal) or 3% (suboptimal) fetal calf serum. Among the different growth factors tested, only epidermal growth factor and to a certain extent insulin had a growth-promoting effect by themselves. The other growth factors had either an additive effect in the presence of epidermal growth factor or no effect at all. The cytotrophoblast cells expressed both epidermal growth factor and transferrin receptors whereas the more differentiated giant cells expressed only transferrin receptors.

Nature of the Multipotential Cells in the Displaced Rat Yolk Sac

The nature and origin of the proliferating cells were studied in displaced visceral yolk sac in vivo and in organ culture in vitro. By ultrastructural examination and reactivity with antibodies against intermediate filaments it was shown that the poorly differentiated cells which appear in this membrane have features in common with visceral endodermal cells. Using plant lectins reacting with the rat visceral endoderm (DBA, PNA, SJA and BSA-1) it was demonstrated that the proliferating endodermal cells retrodifferentiate to earlier stages of endodermal development.