Concentrations Of Fetal Serum Research Articles

Interleukin-1 stimulation of equine articular cells

Prostaglandin E 2 ( pge 2) and stromelysin are produced by equine chondrocytes and synovial cells in vitro in response to recombinant human (rh) interleukin-1 (IL-1) α and β, and equine mononuclear cell supernatants ( mcs) containing IL-1. However, culture conditions are important. pge 2 concentrations increase in proportion to the concentration of fetal calf serum ( fcs) in the culture medium, whereas stromelysin concentrations are inversely proportional to the concentration of fcs. Equine mcs, containing a lower concentration of IL-1 than the concentration of rhIL-1 used in these experiments, stimulated production of much higher levels of pge 2 than rhIL-1. In addition, equine mcs induced the production of broadly similar levels of pge 2 by both chondrocytes and synovial cells, whereas rhIL-1 was more active on equine synovial cells than equine chondrocytes. Although equine Mcs induced both stromelysin and PGEZ production by equine articular cells, on the whole rhIL-1 failed to induce stromelysin production. This supports previous observations of species restrictions in the activity of human IL-1 on equine cells. Therefore, experiments using mammalian cells and heterologous IL-1 should be interpreted with caution.

Onset of spontaneous labor and changes in E2 and progesterone levels--possible interrelation.

Unlike the findings in animal studies, in which a decline in progesterone levels is clearly associated with the onset of labor, investigation of progesterone levels among human parturients has resulted in controversy. This study was designed to address the issue and evaluate labor-onset related changes of estradiol-progesterone (E2-P) concentration in fetal scalp serum, umbilical vein serum and in the peripheral maternal serum. Seven women in spontaneous labor, were compared to 7 women in whom labor was induced. Our results reveal a significant decrease in the maternal serum P concentration when spontaneous labor is taking place (120.6 +/- 24.5 mg/ml verus 177.3 +/- 61.4 mg/ml, p < 0.05). Significant change in the ratio of the fetal scalp to the maternal serum E2/P ratio in women at spontaneous labor versus induced labor is also shown. We could not demonstrate any changes in the E2 levels in relation to labor. We conclude that the onset of labor in human pregnancy is most probably preceded by local changes in the levels of P and ratio of P to E2. These changes may play an important regulatory role in onset of labor.

Effects of adenine nucleotides on the proliferation of aortic endothelial cells.

The effects of adenine nucleotides and adenosine on DNA synthesis and cell growth have been studied in bovine aortic endothelial cells (BAECs). ATP produced a small but significant (+44%) increase of the fraction of BAECs whose nuclei are labeled by [3H]thymidine. This mitogenic effect was mimicked by ADP, the phosphorothioate analogues ATP gamma S and ADP beta S, and the nonhydrolyzable analogue adenosine 5'-(beta, gamma-imido)triphosphate (APPNP), whereas adenosine 5'-(alpha, beta-methylene)triphosphate (APCPP), a selective agonist of P2x-purinoceptors, had no effect at 10 microM and a small one at 100 microM; this profile is consistent with the involvement of P2y-receptors. Adenosine induced a mitogenic response of a magnitude similar to that of ATP. This effect was not reproduced by R-phenylisopropyl adenosine, by 5'-N-ethylcarboxamide adenosine, or by 2',5'-dideoxyadenosine, selective ligands of the A1- and A2-receptors and the P site, respectively, nor was it inhibited by 8-phenyltheophylline, an antagonist of both A1- and A2-receptors. The mechanism of this adenosine action thus remains unclear. ATP and ATP gamma S did not enhance the proliferation of BAECs cultured in the presence of fetal calf serum concentrations ranging from 0.5% to 10%. They inhibited the growth-promoting effect of basic fibroblast growth factor; among the various nucleotides tested, APCPP was the least effective to reproduce the action of ATP, suggesting the possible involvement of P2y-receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Folate and vitamin B12 concentrations in maternal and fetal blood, and amniotic fluid in second trimester pregnancies complicated by neural tube defects.

To investigate maternal and fetal folate and vitamin B12 concentrations in pregnancies affected by neural tube defects (NTD). Measurement of folate and vitamin B12 concentrations in amniotic fluid, fetal blood and maternal blood samples in midgestation. 32 women undergoing termination of pregnancy at 14-21 weeks gestation for social reasons (n = 24) or for fetuses with neural tube defects (n = 8). Fetoscopy before intra-amniotic injection of prostaglandins. In normal pregnancies there was a positive correlation between maternal and fetal serum folate, and the fetal serum and red blood cell folate concentrations were higher than the maternal. There were no differences in amniotic fluid, maternal blood or fetal blood folate concentrations between pregnancies with NTD and normal pregnancies. Although amniotic fluid vitamin B12 was lower in pregnancies with NTD, maternal serum vitamin B12 concentration was not reduced. In this small group of pregnancies with NTD at mid-gestation there is no evidence to suggest folate or vitamin B12 deficiency.

Correlation between nucleolar organizer region score and bromodeoxyuridine labeling index in C6 glioma cell line.

Correlation between the mean number of nucleolar organizer region-associated argyrophilic proteins (AgNORs) per cell and BrdU labeling index (LI), the percentage of S-phase cells, was investigated in vitro using rat C6 glioma cells under several different conditions such as increased concentration of fetal calf serum (FCS) in a stepwise manner in medium. In addition, the effect of dibutyryladenosine cyclic monophosphate (d-cAMP) on the mean number of AgNORs and the BrdU LI was examined. C6 glioma cells cultured with no FCS showed 5.1% BrdU LI and 3.46 AgNOR score. BrdU LI and AgNOR score of the cells cultured with 1% FCS were 26.5% and 6.25, respectively. Those with 5% FCS were 44.7% and 8.41, with 10% FCS; 49.8% and 10.85, and with 20% FCS; 48.4% and 10.64. Treatment of d-cAMP with 10% FCS induced morphological differentiation and reduced BrdU LI (9.4%) and AgNORs score (4.90), compared with C6 cells cultured in medium containing 10% FCS without d-cAMP. Thus, the BrdU LI and AgNOR score of the C6 cells cultured under several different concentration of FCS were shown to be well linearly related (r = 0.97) and both values were decreased with morphological alteration of C6 glioma cells in the presence of 1 mM d-cAMP.

Growth hormone and insulin-like growth factor I regulate collagen gene expression and extracellular collagen in cultures of avian skin fibroblasts

Avian skin fibroblasts were isolated, cultured and incubated with [ 3H]proline for 24 h. The cells exported radiolabeled collagenase-digestible (CDP) and non-collagenase-digestible (NCDP) proteins into the medium. Human, bovine and avian growth hormone (GH) as well as insulin-like growth factor I (IGF-I) attenuated the appearance of [ 3H]CDP in the medium without affecting [ 3H]NCDP. The appearance of [ 3H]CDP was not affected by prolactin. The effects of GH and IGF-I were enhanced by increasing concentrations of fetal calf serum (FCS). A synergism was observed between GH and IGF-I in their effect on CDP. Each peptide, at an ineffective concentration, increased the sensitivity of the cells to the other peptide. Collagenase activity in the medium was enhanced by IGF-I, but not modified by GH, FCS, or by their interaction with IGF-I. GH and IGF-I inhibition of type I procollagen gene expression was demonstrated with the aid of probes containing sequences corresponding to the mRNAs for avian αI and αII chains. The results suggest that GH and IGF-I cooperate in regulating collagen synthesis, but collagen degradation is affected by IGF-I and not by GH.

Increased Collagen Gene Expression in Vascular Smooth Muscle Cells Cultured in Serum or Isoleucine Deprived Medium

The relationship between type I and type III collagen gene expression and the growth state of cultured bovine vascular smooth muscle cells has been investigated. Growth was modulated by incubation of subconfluent cultures of smooth muscle cells in media containing concentrations of fetal calf serum ranging from 10% to 0.5%. As the serum concentration was lowered, the doubling time increased from 17 hours to 70 hours, and collagen mRNA levels increased as judged by Northern blot analysis. The levels of induction were 5- to 15-fold for al (1), 2- to 3-fold for a2(I), and 4- to 7-fold for a1(III). Metabolic cell labelling with precursor amino acids indicated that type l and type III procollagen synthesis was elevated approximately 2-fold as growth slowed. A similar 2-fold increase occurred upon translation of isolated RNAs in a rabbit reticulocyte cell-free system. Nuclear run-on analysis indicated that increased collagen transcription can account for part of the increase in mRNA levels following incubation in lowered fetal calf serum. To further examine the influence of proliferation on collagen gene expression, smooth muscle cells were cultured in isoleucine-free medium, which resulted in quiescence within 36 hours. Induction of collagen mRNA levels was observed within 24 hours of incubation in isoleucine-free medium. Thus, collagen gene expression increases as the growth of vascular smooth muscle cells is slowed by either deprivation of growth factors or essential amino acids. Regulation is mediated at several sites, including gene transcription and mRNA translation, and possibly post-transcriptional steps.

Autocrine secretion of an EGF-like substance by a cell line (HSC-1) derived from a human skin squamous cell carcinoma

The presence of EGF-autocrine secretion was investigated in a cell line derived from a human squamous cell carcinoma (HSC-1). The cell line was found to secrete the epidermal growth factor (EGF)-like substance. Meanwhile, normal human and uninvolved and involved psoriatic epidermal cells did not show any evidence of EGF secretion in culture. Not was any evidence of EGF secretion observed in vitro in several malignant cell lines other than HSC-1 derived from human squamous cell carcinomas, adenocarcinoma and melanoma. The addition of EGF did not stimulate, but rather inhibited, the growth of HSC-1 cells in GIT medium as well as Dulbecco's modified essential medium with low concentrations of fetal calf serum (0.5–1%) in vitro. Overexpression of EGF receptors is known to occur in HSC-1. The results suggest that HSC-1 cells exhibit autocrine secretion of the EGF-like substance but not autostimulation in anchorage-dependent cell culture.

MULTIPLE GROWTH ABNORMALITIES IN VASCULAR SMOOTH MUSCLE FROM SPONTANEOUSLY HYPERTENSIVE RATS

1. In tissue culture the growth characteristics of aortic smooth muscle cells isolated from spontaneously hypertensive rats (SHR) were compared with those of normotensive Wistar-Kyoto (WKY) rats. 2. Aortic smooth muscle cells from SHR exhibit enhanced proliferation when grown in the presence of low (1%) and moderate (5%) concentrations of fetal calf serum. 3. Cell quiescence in cultures of smooth muscle from SHR becomes apparent at cell densities approximately 20% higher than in cultures from WKY rats. 4. These different growth characteristics of smooth muscle between the two strains of rats may contribute to the early pre-hypertensive development of vascular hypertrophy in the SHR.

Receptor-mediated endocytosis of albumin in cultured opossum kidney cells: a model for proximal tubular protein reabsorption.

The accumulation of fluorescein (FITC)-labelled bovine albumin was measured against the extracellular-fluid-phase marker FITC-inulin within confluent monolayers of the opossum kidney cell line OK. Fluorescence and electron microscopic pictures show that FITC-albumin is taken up by endocytosis and appears in a vesicular intracellular distribution. The uptake of FITC-albumin was quantified by measuring the cell-adherent fluorescence fluorimetrically. FITC-albumin uptake shows a time- and concentration-dependent saturation kinetics in contrast to the non-saturable FITC-inulin uptake, and exceeds the latter more than tenfold at low concentrations. Half-maximum saturation occurs at 20-30 mg/l. Initial FITC-albumin uptake/mg protein is stimulated by cell maturation, being six-to sevenfold higher in the confluent than in the subconfluent state, while FITC-inulin uptake is unchanged. Both an elevation of ambient osmolality to 600-750 mOsm/kg and disruption of the cytoskeleton by cytochalasin B (0.1 mmol/l) reduce initial FITC-albumin uptake by 50%-60% in a non-additive fashion. Albumin endocytosis is reduced both in acidic (pH 5.4) and alkaline (pH 8.4) medium, but does not depend on extracellular sodium, calcium or chloride. High concentrations of fetal calf serum or unlabelled albumin reduce FITC-albumin endocytosis dose-dependently. The present study is the first to investigate both the protein uptake and the fluid-phase endocytosis in a cultured proximal tubular cell line, using these cells as a model system-for proximal tubular protein reabsorption.

Maturation of the Secretion of Thyroid Hormone and Thyroid-Stimulating Hormone in the Fetus

Data on human fetal thyroid function have largely been derived from histologic studies or studies of cord-blood samples obtained at hysterotomy or delivery. These data may not represent true normal values. Cordocentesis (ultrasound-guided blood sampling from the umbilical cord) is a technique that allows investigation of physiologic processes in fetuses not under stress. We measured serum thyroid-stimulating hormone, total and free thyroxine (T4), total and free triiodothyronine (T3), and thyroxine-binding globulin in blood samples from 62 fetuses. The samples were obtained by cordocentesis (n = 58) or cardiocentesis (n = 4) at 12 to 37 weeks of gestation. Maternal serum samples were obtained immediately before fetal blood sampling. Fetal serum thyroid-stimulating hormone, thyroxine-binding globulin, and total and free T4 and T3 concentrations increased significantly with the length of gestation (P less than 0.001). The only significant association among these variables, independent of the length of gestation, was between thyroid-stimulating hormone and free T4 (P less than 0.0001). Maternal serum concentrations of these variables did not change during gestation, and there was no significant relation between fetal and maternal values. Most fetal serum concentrations of thyroid-stimulating hormone were higher, whereas most serum total and free T3 concentrations were lower than the respective values for normal adults. The fetal serum total T4, free T4, and thyroxine-binding globulin values reached the level of the mean adult values at approximately 36 weeks of gestation. The increases in fetal serum concentrations of thyroid-stimulating hormone, thyroxine-binding globulin, and total and free T4 and T3 during gestation reflect increasing maturation of the pituitary, thyroid, and liver. The finding of increasing fetal serum concentrations of thyroid-stimulating hormone in the presence of increasing thyroid hormone concentrations suggests that the sensitivity of the fetal pituitary gland to negative feedback is limited or is counterbalanced by increasing stimulation by thyrotropin-releasing hormone from the hypothalamus.

Enhancement of Murine Erythopoiesis in Vitro by a Porcine Kidney Extract.

We have examined the effect of porcine kidney extract (PKE) on the growth of erythroid progenitor-derived colonies in a methylcellulose culture. The addition of PKE resulted in an enhancement of burst-forming-unit-erythroid (BFU-E)-derived colonies, and the enhancement of the colony was also observed in a low concentration of erythropoietin (Epo) and fetal calf serum (FCS). The activity of PKE on BFU-E was compared with the erythroid growth factors, such as recombinant murine interleukin-3 (IL-3), recombinant murine granulocyte/macrophage colony-stimulating factor (GM-CSF) and recombinant human granulocyte colony-stimulating factor (G-CSF) which are all well known to stimulate BFU-E growth. IL-3 showed a potent burst-promoting activity (BPA), but GM-CSF and G-CSF did not enhance BFU-E growth in our culture conditions. In the cultures supplemented with Epo, the rapid loss of BFU-E was prevented with IL-3; however, PKE alone did not prevent the disappearance of BFU-E. These results suggest that PKE possesses a BPA-like activity which is considered an enhancement of BFU-E.

Use of cryopreserved animal and human hepatocytes for cytotoxicity studies

To evaluate the potential use of cryopreserved hepatocytes for toxicological studies, rat, dog and human hepatocytes were frozen in Leibovitz medium containing 20% foetal calf serum and various concentrations of dimethylsulphoxide and stored in liquid nitrogen. 50% or more of the hepatocytes that attached and survived immediately after isolation still possessed these properties after freezing/thawing. Thawed hepatocytes from the three species showed only a slight reduction in their ability to metabolize phenacetin or to conjugate paracetamol with glucuronic acid. Sensitivity to the toxic effects of erythromycin was not affected by the MTT and neutral red assays. Similar observations were made with rat hepatocytes for five other toxic agents—chloramphenicol, chlorpromazine, acrylamide, chloroquine sulphate and p-chloromercuribenzoic acid. These results suggest that, after cryopreservation, isolated hepatocytes represent a suitable model for drug metabolism and toxicity screening studies.

Placental transfer of cefazolin and piperacillin in pregnancies remote from term complicated by Rh isoimmunization

Although the administration of prophylactic antibodies for intrauterine transfusion is controversial, little information is available regarding placental transfer of antibiotics administered to the mother, or whether the presence of hydrops affects this placental transfer. Sixteen intravascular intrauterine transfusions were performed in 10 patients. Seven (10 procedures) patients were given 2 gm of cefazolin before the procedure and samples were obtained by fetal vascular access. Three patients (six procedures) were given 4 gm of piperacillin and samples were similarly obtained. Specimens were obtained for fetal serum, maternal serum, and amniotic fluid antibiotic concentration. The mean serum cefazolin concentration in hydropic fetuses was 18.04 ± 3.37 (ig/ml, and in nonhydropic fetuses the concentration was 21.02 ± 17.8 µg/ml (p = 0.72). The mean fetal serum concentration of piperacillin was 22 ± 12 (xg/ml. The placental transfer of both drugs was similar. We conclude that the transplacental passage of these antibiotics is prompt and that the presence of hydrops does not significantly impair the passage of cefazolin.

Perinatal Growth Hormone (GH) Physiology: Effect of GH-Releasing Factor on Maternal and Fetal Secretion of Pituitary and Placental GH*

To study regulation of the secretion of human pituitary GH (hGH) and placental GH (hPGH) in the pregnant woman and human fetus, the GH-releasing factor Sermorelin [GRF-(1-29)-NH2] was administered to pregnant women at term (n = 5), just before elective cesarean section; saline was administered in control studies (n = 5). The effects of GRF-(1-29)-NH2 administration on maternal and fetal serum concentrations of hGH and GRF-(1-29)-NH2 and maternal serum levels of hPGH were evaluated at birth. The mean time span between injection and birth was 20 min (range, 15-25 min). Cord serum hGH concentrations were similar in infants of GRF-(1-29)-NH2-injected mothers and control infants. GRF-(1-29)-NH2 elicited a consistent but small rise in maternal hGH serum concentrations (P = 0.08), whereas hPGH concentrations remained unaltered. Finally, GRF-(1-29)-NH2 concentrations were undetectable in cord serum, but readily detectable in concomitantly obtained maternal serum. In conclusion, these data suggest that hGH secretion in the pregnant woman at term is suppressed at the pituitary level, that GRF does not affect hPGH secretion, and that fetal hGH secretion is independent of circulating maternal GRF, probably because of lack of transplacental GRF passage.

PIG FETAL PANCREATIC MONOLAYERS

Endocrine-rich monolayers of pig fetal pancreas that are free of fibroblasts have been established with the ultimate aim of providing guidelines for the culture of the human equivalent. The immunogenic potential of the monolayers--hence their capacity to be grafted--has also been analyzed. Fetuses ranging from 50 to 90 days were used, and, following digestion with collagenase (4 mg/ml, 15-20 min), the pancreatic suspension was plated onto tissue culture vessels containing RPMI 1640. The fetal calf serum concentration was kept low (5%) initially to inhibit fibroblast proliferation, but subsequently increased to 7%. Monolayers from a typical litter of 8-10 fetal pigs produced 6-8 x 10(8) viable epithelial cells by day 10 of culture, of which 75% were endocrine cells. This represents an 8-fold increase in a two-week period. The ratio of beta:alpha:delta:pancreatic polypeptide cells was 19:33:18:5. These monolayers synthesized both DNA, (pro)insulin and protein, and displayed increased insulin release when exposed to 10 mM theophylline, 10 mM Ca2+ and 1.3 microM 12-0-tetradecanoyl-phorbol-13-acetate. Static stimulation with 20 mM glucose however, did not elicit a response in insulin secretion. These cells displayed no reaction to allogeneic lymphocytes in a mixed lymphocyte culture, whereas freshly obtained porcine epithelial cells did. Methods may need to be found to increase the proportion of B cells in this enriched endocrine cell population. In general however, guidelines have been established that may be useful in developing a monolayer of human fetal pancreatic cells with the eventual aim of transplantation. The reduction in immunogenicity of the pig fetal pancreatic cells suggests that they too might be a potential source for transplantation.

Prostaglandins and rat glomerular mesangial cell proliferation

Arachidonate metabolites modulate glomerular mesangial cell contractility through specific receptors coupled to phospholipase C or adenylate cyclase. The resulting intracellular signals, including changes of cytosolic Ca2+, pH, and cyclic adenosine 3'5'-monophosphate (cAMP) are known to also regulate the growth of many cell types. Since eicosanoids have been shown to interfere with cell proliferation in culture, we studied DNA synthesis and cell number in rat mesangial cell cultures exposed to a selective phospholipase C activator, prostaglandin F2 alpha (PGF2 alpha), or to the cAMP-stimulating PGI2 analogue, Iloprost. PGF2 alpha dose-dependently enhanced DNA synthesis and cell proliferation in the presence of insulin, with an EC50 of 0.1 microM. This eicosanoid potentiated the effects of platelet-derived growth factor (PDGF) or low concentrations of serum. Maximum stimulatory potency was about one-third that of PDGF. Removal of PGF2 alpha after short-term stimulation (30 min) did not reverse its mitogenic effect. Iloprost had no effect on DNA synthesis of quiescent cells, but potently inhibited growth stimulated by various concentrations of fetal serum. PG released within the glomerular microcirculation may play a regulatory role in both normal and deranged mesangial cell growth.

Intra- and extracellular proteins in human normal and polycystic kidney epithelial cells

A tissue culture method was established for the continuous growth of epithelial cells from the cortex of human normal kidney (HNC) and from the epithelial layer of kidney cysts from autosomal dominant polycystic kidney disease (ADPKD) patients. Primary cells were grown to 80 to 90% confluency from 1 mm2 slices of tissue, and subcultured up to 10 times. The subcultured HNC and ADPKD cells retained characteristic epithelial polygonal and elongated shape and positive immunofluorescent staining for cytokeratin. The cell doubling time for both HNC and ADPKD epithelia was three to four days at a fetal calf serum (FCS) concentration of 5%. Using these culturing procedures 1 to 5 x 10(9) epithelial cells could be obtained from each kidney specimen. Profiles of 35S-methionine radiolabeled intracellular proteins of HNC and ADPKD cells qualitatively demonstrated a high degree of similarity, thus confirming a similarity of epithelial origin and protein biosynthesis. Both the underexpression of three proteins (a) protein p2, Mr approximately 47 kDa, pI approximately 6.0; b) protein p3, Mr approximately 50 kDa, pI approximately 5.9; and c) protein p4, Mr approximately 44 kDa, pI approximately 5.8) and the overexpression of several proteins (including: a) p5, Mr approximately 56 kDa, pI approximately 7.3; b) protein p6, Mr approximately 32 kDa, pI approximately 7.3; c) protein p7, Mr approximately 33 kDa, pI approximately 5.3; d) protein p8, Mr approximately 45 kDa, pI approximately 6.9; e) protein p9, Mr approximately 35 kDa, pI approximately 6.7; and f) protein p10, Mr approximately 30 kDa, pI approximately 6.6) were found in ADPKD cells.(ABSTRACT TRUNCATED AT 250 WORDS)

The neuropeptide histidyl proline diketopiperazine throughout human pregnancy: An inverse correlation with amniotic fluid prolactin

Histidyl proline diketopiperazine values have been established in human amniotic fluid (n = 81) and maternal serum (n = 36) throughout gestation (10 to 42 weeks). Newborn cord serum (n = 10) and first-voided fetal urine (n = 10) levels were also documented. These measurements reveal increasing amniotic fluid levels with term gestation values (15,551 pg/ml) nearly thirteen-fold higher than maternal serum concentrations (1150 pg/ml). Corresponding fetal urine and cord serum concentrations were 16,781 and 2160 pg/ml, respectively. The amniotic fluid values are not influenced by fetal sex or maternal labor, nor do they correlate with amniotic fluid a-fetoprotein levels. However, there is a significant inverse correlation (r = -0.628; p<0.0001) between amniotic fluid prolactin and histidyl proline diketopiperazine after midgestation. The hypothesis that histidyl proline diketopiperazine may be a regulatory peptide for decidual prolactin production was tested by culturing term decidua in the presence of varying concentrations of histidyl proline diketopiperazine, but no inhibitory effect was observed. Decidual cultures did not produce measurable amounts of histidyl proline diketopiperazine. It is suggested that amniotic fluid histidyl proline diketopiperazine is derived from fetal urine. (AM J OesTET GYNECOL 1990;162:740-5.)

Active Oxygen‐mediated Cytotoxic and Antitumor Actions of Streptococcal Cytotoxic Protein

Streptococcal cytotoxic protein (SCP), obtained from the cell‐free extract of Streptococcus pyogenes, inhibited the uptake of [methyl‐3H]thymidine by Ehrlich ascites carcinoma cells depending on the concentration of fetal calf serum (FCS) added to the culture medium. The same results were found using sera from different animals. The inhibitory activity of SCP was completely suppressed by catalase but not by hydroxyl radical scavengers or superoxide dismutase. The antitumor activity of SCP in tumor‐bearing mice was inhibited by administration of catalase together with SCP. SCP enzymatically produced hydrogen peroxide in the presence of FCS as detected by the 2,2′‐azino‐di‐[3‐ethyl‐benzothiazoline‐(6)‐sulfonic acid]/horseradish peroxidase method. The kinetic parameters, Km and Vmax, for hydrogen peroxide production of SCP were 2.13% FCS and 0.53 nmol/min/μg SCP, respectively. These results indicate that SCP is an enzyme which produces hydrogen peroxide and exerts potent cytotoxic and antitumor effects using an active oxygen species, hydrogen peroxide, produced by the enzymatic reaction of SCP with an unknown substrate contained in FCS or sera from various animals and molecular oxygen.