Spreds, inhibitors of the Ras/ERK signal transduction, are dysregulated in human hepatocellular carcinoma and linked to the malignant phenotype of tumors.

Aberrant activation of the Ras/Raf-1/extracellular-regulated kinase (ERK) pathway has been shown to be involved in the progression of human hepatocellular carcinoma (HCC). However, the mechanism of dysregulation of ERK activation is poorly understood. Recently, we identified Sprouty-related protein with Ena/vasodilator-stimulated phosphoprotein homology-1 domain (Spred) as a physiological inhibitor of the Ras/Raf-1/ERK pathway. In this study, we found that the expression levels of Spred-1 and -2 in human HCC tissue were frequently decreased, comparing with those in adjacent non-tumorous tissue. Moreover, Spred expression levels in HCC tissue were inversely correlated with the incidence of tumor invasion and metastasis. Forced expression of Spred-1 inhibited HCC cell proliferation in vitro and in vivo, which was associated with reduced ERK activation. Spred-1 overexpression also reduced the secretion of matrix metalloproteinase-9 (MMP-9) and MMP-2, which play important roles in tumor invasion and metastasis. In addition, Spred-1 inhibited growth factor-mediated HCC cell motility. These data indicate that the reduction of Spred expression in HCC is one of the causes of the acquisition of malignant features. Thus, Spred could be not only a novel prognostic factor but also a new therapeutic target for human HCC.

Adduct of acetylene at sulfur in an oxygen- and sulfur-bridged open cubane cluster complex of tungsten.

Formation of a single carbon-sulfur bond is described. The reaction of incomplete cubane-type sulfur and oxygen-bridged isothiocyanato tungsten cluster [W3(mu3-S)(mu-O)(mu-S)2(NCS)9]5- (7) with acetylene affords [W3(mu3-S)(mu-O)(mu-S)(mu-SCH=CH2)(NCS)9]4- (8). The cluster 8 has been isolated as K0.5(Hpy)3.5[W3(mu3-S)(mu-O)(mu-S)(mu-SCH=CH2)(NCS)9] (8'), whose structure has been characterized by X-ray crystallography, electronic spectra, and 1H and 13C NMR spectroscopy. Crystal data of 8': triclinic system, space group P1, a = 14.465(5) A, b = 17.353(3) A, c = 10.202(2) A, alpha = 90.98(1) degrees, beta = 108.59(2) degrees, gamma = 98.13(2) degrees, V = 2397.6(10) A(3), Z = 2, D(c) = 2.096 g cm(-3), Dm = 2.08 g cm(-3), R (Rw) = 3.6 (5.5)% for 8786 reflections (I > 1.50 sigma(I)). The carbon-carbon distance is 1.27(1) A and is almost equidistant between ethylene (1.339 A) and acetylene (1.203 A). The electronic spectrum of 8' in 1.0 M HCl containing 1.5 M KSCN has a characteristic broad peak in the near-infrared region [lambda(max), nm (epsilon, M(-1) cm(-1)): 840 (650), 575 (1450)]. (1)H NMR and HH correlation spectroscopy (COSY) of 8' in CD3CN support the results of the X-ray structural analysis. The (1)H NMR spectrum shows three signals at 2.42 (1H, dd), 4.84 (1H, d, J = 8.8 Hz), and 4.89 (1H, d, J = 16.2 Hz) ppm due to the mu-SCH=CH2 moiety of 8'. The correlation spectrum shows spin couplings of the signal at 2.42 ppm with the signals at 4.84 and 4.89 ppm. The mechanism of the formation of 8 is suggested to proceed through an intermediate with acetylene bridging two of the sulfur atoms.

[An outbreak of Pseudomonas sepsis associated with nosocomial infection in a pediatric ward].

A five-year old girl, diagnosed with fever of unknown origin (FUO), was transferred to our hospital due to a deterioration in her general condition. Pseudomonas cepacia (P. cepacia) was detected in her blood culture and she soon recovered after imipenem cilastatin (IPM/CS) administration. When the former hospital was informed of the results of her blood cultures, we learned that five other cases of FUO had occurred there within the previous two weeks in noncompromised children. Except for one case, Pseudomonas species was detected in their blood cultures. P. cepacia was found in one case, Pseudomonas aeruginosa (P. aeruginosa) was detected in two other cases, and both P. cepacia and P. aeruginosa were demonstrated in the other patient. Although the origin of the bacteria is unknown, it may have spread to the children through a contaminated liquid reservoir or medical devices.

Effects of androgen, fibroblast growth factors or other various growth factors on growth of Shionogi carcinoma cells in a protein-free medium.

Shionogi carcinoma 115 (SC115) has been accepted for 20 years as an androgen-responsive mouse mammary tumor. We have recently established an androgen-dependent cloned cell line (SC-3) from a SC115 tumor. We found in the present study that the growth of SC-3 cells can be stimulated by 10(-8) M testosterone (up to 100-fold) even in a protein-free medium beginning from plating [Ham's F-12: Eagle's minimum essential medium (1:1, v/v)]. In the protein-free culture, the proliferation of SC-3 cells was also found to be stimulated by acidic or basic fibroblast growth factor (FGF) alone (up to 50-fold) among various growth factors examined such as FGFs, insulin, insulin-like growth factor (IGF)-I, IGF-II, nerve growth factor, platelet-derived growth factor, epidermal growth factor, transforming growth factor (TGF)-alpha and TGF-beta. The testosterone (10(-8) M)- or FGF (10 ng/ml)-induced growth of SC-3 cells was abolished only by TGF-beta (greater than or equal to 1 ng/ml) among various growth factors examined. We show for the first time in this study an androgen-dependent growth of cancer cells (SC-3) in a protein-free medium. In the protein-free medium, growth of SC-3 cells is also stimulated by FGFs, and the androgen- or FGF-induced growth is inhibited only by TGF-beta.

Growth stimulation by androgens, glucocorticoids or fibroblast growth factors and the blocking of the stimulated growth by antibody against basic fibroblast growth factor in protein-free culture of Shionogi carcinoma 115 cells.

Shionogi carcinoma 115 (SC115) has been accepted for 20 years as an androgen-responsive mouse mammary tumor. We have established an androgen-dependent cloned cell line (SC-3) from a SC115 tumor. In a serum-free medium, testosterone (T) or fibroblast growth factors (FGFs) markedly stimulate the growth of SC-3 cells, and the T-induced growth was shown to be mediated through FGF-like peptide(s) in an autocrine mechanism. Since we used the serum-free culture including 0.1% bovine serum albumin (BSA), a partially serum-containing condition, putative roles of BSA- or serum-borne growth factors in growth stimulation of autocrine production of FGF-like peptide(s) could not be excluded. This paper reports findings performed in a protein-free medium including plating [Ham's F-12:MEM (1:1; v/v)]. In the protein-free culture, the growth of SC-3 cells was significantly stimulated by the addition of greater than or equal to 10(-10) M T (up to 20-fold), greater than or equal to 10(-7) M dexamethasone (Dex; up to 7-fold) or greater than or equal to 1 ng/ml basic (b) or acidic FGF (up to 10-fold); other various growth factors had no such effects. Furthermore, DNA synthesis of SC-3 cells induced by T, Dex or bFGF was similarly and markedly inhibited by bFGF neutralizing antibody IgG. Therefore, the present findings seem to demonstrate that androgens or high levels of glucocorticoids induce the production and secretion of FGF-like peptide(s) from SC-3 cells for their growth even in the absence of additional support by other factors.

Uterine papillary serous carcinoma with high levels of serum carcinoembryonic antigen. Response to combination chemotherapy.

A case of metastatic uterine papillary carcinoma with elevated serum carcinoembryonic antigen (CEA) is reported. The patient presented with ascites and pleural effusions and with a high level of CEA (258 ng/ml) 3 months after primary surgical resection and postoperative irradiation had been performed. Her complete response, although temporary, to two kinds of combination chemotherapy was evaluated using serial estimations of the serum CEA.

The effects of A23187 and phorbol ester on the phosphorylation of proteins in rat anterior pituitary cells.

The effects of A23187 and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on the phosphorylation of proteins in normal rat anterior pituitary cells were compared. A23187 rapidly activated the phosphorylation of at least 5 proteins (45, 47, 53, 54 and 58 kDa), which reached the maximal level in 2-10 min, and decreased gradually thereafter. In contrast, TPA activated the phosphorylation of at least 6 proteins (45, 62, 64, 72, 76 and 82 kDa), which were mostly distinguished from those activated by A23187. TPA-induced response was elicited more slowly, reached a plateau after about 10 min, and was sustained thereafter. These results suggest that the protein phosphorylation stimulated by A23187 and TPA is conducted by different mechanisms.

Small cell carcinoma of the endometrium associated with adenosquamous carcinoma: a light and electron microscopic study.

This report describes a 64-year-old woman with a primary small cell carcinoma of the endometrium associated with adenosquamous carcinoma. The light microscopic features resembled those of small cell carcinoma of the lung and those of the uterine cervix, and foci of adenosquamous carcinoma lay scattered sparsely in the small cell carcinoma. Electron microscopy revealed cytoplasmic neurosecretory type granules. The neoplasm behaved in a very aggressive manner such that at 3 months after surgery a metastatic neoplasm appeared in the vagina. This case is a rare example of an endometrial carcinoma with differentiation toward endocrine as well as adeno and squamous cell carcinoma.

[Clinical studies of 145 cases of endometrial carcinoma--analysis of prognostic factors, especially pathological types].

One hundred and forty-five patients with confirmed endometrial carcinoma treated at Kumamoto University Hospital were studied from the clinicopathological point of view. As risk factors, the incidence of high age, postmenopause, atypical genital bleeding, sterility, nulliparity, diabetes mellitus, hypertension and obesity was showed to be high. These factors may be expected to assist in screening for early diagnosis by using each factor or combination of factors. As prognostic factors, we examined the relationship between stage, depth of myometrial invasion, histological grade, histological type and survival rate. The stage was shown to be a most important predicator of survival. The depth of myometrial invasion and the histological grade closely correlated with the stage. The relationship between the histological subtype, especially papillary serous carcinoma (PSC) and prognosis, was investigated in our series of studies. PSC found in an incidence of 9.4% had a relatively poor prognosis compared with endometrioid carcinoma. Its 5-year survival rate was only 30%. Because PSC has a potential for aggressive invasion and rapid metastasis as compared to endometrioid carcinoma, careful histological examination and aggressive treatment are warranted.

Immunological studies of human endometrium--suppression of lymphocyte mitogenesis by endometrial extract.

Immunoregulatory activity that suppresses lymphocyte mitogenesis was demonstrated in human secretory endometrial extracts (SEE). SEE markedly suppressed 3H-thymidine (3H-TdR) incorporation by phytohemagglutinin (PHA)-stimulated lymphocytes compared with myometrial extracts from the same donors. SEE-induced inhibition of lymphocyte response to PHA was significantly (p less than 0.01) greater than that in cultures containing proliferative endometrial extracts at equivalent concentrations. SEE inhibited PHA- and ConA-stimulated cultures much better than PWM cultures, suggesting that it suppressed mitogen stimulation of T cells, but probably not B cells. Investigation of the kinetics of SEE-mediated suppression revealed that it was effective in inhibiting the initial activation processes of T-cell mitogenesis. Preincubation of lymphocytes with SEE followed by washing prevented inhibition of PHA-stimulated proliferation, indicating that the inhibitory effect of SEE might be reversible. The results are discussed with respect to a possible role of the inhibitory factor(s) in SEE in facilitating the implantation process of fetal allograft.

Clear cell carcinoma and endometrioid carcinoma arising in a endometriotic cyst of the ovary.

A case of two tumors arising in an endometriotic cyst of the ovary is described. These two tumors are macroscopically polypous and are histologically a clear cell carcinoma and an endometrioid carcinoma, respectively. This is a rare neoplasm and its existence supports the general opinion that clear cell carcinoma and endometrioid carcinoma of the ovary are both müllerian in origin.

Fetal blood velocity waveforms in uncomplicated pregnancy.

Umbilical artery velocity waves were measured in fetuses from 94 normal pregnant women. In all, 183 determinations were carried out from the 14th to the 40th week of gestation. A combination of pulsed echo and real-time scanning was used to obtain blood waveforms from the umbilical arteries. The umbilical artery velocity wave can be readily differentiated from other fetal signals by its pattern. The systolic peak of the velocity wave was divided by the end diastolic value, thereby giving an S/D ratio. The S/D ratio in normal pregnancy declined from 7.6 to 2.0 from 14 to 40 weeks. Analysis of these waveforms indicated that the placenta is an organ of low vascular resistance and that placental resistance to blood flow declines with advancing gestational age in normal pregnancy. The umbilical artery S/D ratio provides a new and non-invasive marker of fetoplacental blood flow resistance.

A case of clear cell adenocarcinoma of the uterine cervix in pregnancy.

An extremely rare case of clear cell adenocarcinoma of the uterine cervix in pregnancy is reported. The primary lesion was first found at 34 weeks of pregnancy in a 34-year-old patient, and was cytologically suspected to be malignant. At 39 weeks of pregnancy, the patient underwent abdominal cesarean section concomitantly with removal of huge right ovarian tumor (3050 g) and delivered a normal male baby weighing 3590 g. Histological findings for cone biopsy at 22 days postpartum revealed invasive adenocarcinoma, and abdominal radical hysterectomy and pelvic lymphoadenectomy were performed at 36 days postpartum. Microscopically, the tumor tissue was composed of clear and hobnail-type cells. It showed a considerable amount of PAS-positive diastase-labile glycogen but was only weakly positive for immunoperoxidase staining of carcinoembryonic antigen.

Inguinal sweat gland carcinoma with choriocarcinomatous differentiation.

An inguinal sweat gland carcinoma is described including the unusual occurrence of alpha hCG. The alpha hCG was demonstrated in the tumor presenting choriocarcinomatous differentiation and undifferentiated polygonal cells in ordinary tumor nests. beta hCG and SP-1 were weakly and sporadically demonstrated in a small area of the tumor presenting choriocarcinomatous differentiation. On the other hand, hPL and histochemical reaction of placental type alkaline phosphatase were not observed in the choriocarcinomatous and undifferentiated tumor cells. CEA was observed in both ordinary sweat gland carcinoma nests and choriocarcinomatous differentiated regions. In the ordinary tumor nests, enzyme histochemical reactions of phosphorylase and SDH were positive. And S 100 protein was sporadically demonstrated. In spite of chemotherapy and radiotherapy, the patient died after a short duration of the disease.

Biochemical and immunohistochemical studies on carcinoembryonic antigen of ovarian mucinous and serous tumors.

The carcinoembryonic antigen (CEA) in the cyst fluid of ovarian mucinous and serous tumors was investigated. The molecular weight and antigenicity of the CEA from both ovarian tumors were very similar to those of colon cancer CEA as determined by SDS electrophoresis and double immunodiffusion on agar plates. In the cyst fluid of ovarian mucinous tumors, the amount of CEA was generally high and CEA of molecular weight (MW) 200,000 was increased. In contrast, in the cyst fluid of ovarian serous tumors, the CEA amount was low and CEA variants of MW 370,000 and 180,000 were present in addition to the main CEA of MW 200,000. Immunohistochemically, CEA was stained mainly in the intestinal type epithelium of ovarian mucinous tumors, and the CEA revealed a tendency to be stained more frequently and strongly with increasing degree of tumor malignancy. Thus, ovarian mucinous tumors (especially the intestinal type epithelium) produced large amounts of CEA which closely resembled colon cancer CEA, whereas ovarian serous tumors produced small amounts of CEA, including some CEA variants. In the study of ovarian epithelial tumors, CEA may be useful as a marker for the malignant transformation of ovarian mucinous tumors.

Significance of angiotensin sensitivity test for prediction of pregnancy-induced hypertension.

In 48 normotensive women with epidemiologic high-risk factors for pregnancy-induced hypertension, the angiotensin sensitivity test was performed serially between 26 and 32 weeks of gestation. If an effective pressor dose of less than 12 ng/kg per minute was considered to be a positive test result, 20 subjects were positive at 30 weeks of gestation and destined to develop pregnancy-induced hypertension. Of ten subjects with a false-positive test result, seven patients developed proteinuria and/or clinically significant edema without apparent hypertension, and only three subjects remained normal throughout their pregnancy. Twenty-eight subjects with negative test results had uneventful pregnancies. Before 30 weeks' gestation, it was difficult to identify all patients destined for pregnancy-induced hypertension. Although high false-positive test results were detected, these results suggest that an angiotensin sensitivity test at 30 weeks' gestation represents an appropriate means of identifying women who remain normal throughout pregnancy. Careful follow-up should be undertaken in all patients with positive test results.