UCI-VULV-1, a vulvar squamous carcinoma cell line.

Squamous carcinoma of the vulva (SCV) is an uncommon neoplasm of uncertain etiology. There is evidence that there are two subgroups of SCV, one associated with human papilloma virus (HPV) and a second HPV-negative group. The UCI-VULV-1 cell line, obtained from a lymph node metastasis of an SCV, grows with a population doubling time of approximately 60 hr. The saturation density is 10(5) cells/cm2. The cell line does not exhibit anchorage independence and is weakly tumorigenic. The cells range in appearance from an abundant spindle cell to a less common larger, flat cell. All of the cells are immunoreactive for high-molecular-weight keratin, but only the flat cells, which form squamous pearls in vivo, are immunoreactive for low-molecular-weight keratin. The cell line expresses epidermal growth factor (EGF), transforming growth factor-alpha, the EGF receptor, and p53 protein. Polymerase chain reaction revealed no HPV DNA within the cells. Early passage cells exhibited karyotypic heterogeneity with few similarities to previous described SCV karyotypes. The cells display sensitivity to cis-platinum in concentrations toxic to many ovarian and cervical carcinoma lines. UCI-VULV-1 may be helpful for studying the properties of the HPV-negative form of SCV.

Does perioperative blood transfusion affect survival in patients with cervical cancer treated with radical hysterectomy?

OBJECTIVE: To determine if blood transfusions during or after radical hysterectomy adversely affect survival in patients with invasive cervical carcinoma. METHODS: Two hundred eighty-three women with stage IA2-IIA cervical cancer were treated with radical hysterectomy and pelvic lymphadenectomy from 1980-1989. Thirteen were lost to follow-up, and five others received adjuvant chemotherapy. Among the remaining 265 patients, 131 were given blood transfusions during surgery or within 30 days, whereas 134 were not. The clinical and pathologic characteristics of these two groups were reviewed and analyzed statistically. RESULTS: Transfused and nontransfused patients did not differ with respect to mean age (45.0 versus 43.4 years, respectively), stage, grade, cell type, depth of invasion, or prevalence of nodal metastasis. Transfused patients more frequently received adjuvant pelvic irradiation than did nontransfused (47 versus 33%, respectively, chi 2 P < .05). After a mean follow-up of 51 months (range 13-125), 19 women (14%) in each group were diagnosed as having recurrent disease, predominantly in the pelvis. Using life-table analysis, the calculated 5-year survival was 86% for transfused and 84% for nontransfused patients, a nonsignificant difference. Disease-free survival was also similar. In the study population, grade, depth of invasion, and nodal status predicted survival. When patients were stratified according to age, cell type, stage, depth of invasion, nodal involvement, and use of adjuvant radiation, blood transfusion still did not adversely influence survival. Using the Cox proportional hazards model, only nodal status was an independent predictor of death. CONCLUSION: Perioperative blood transfusion does not impact overall survival or time to recurrence after radical hysterectomy.

Inhibition of growth of human ovarian cancer in nude mice by luteinizing hormone-releasing hormone antagonist Cetrorelix (SB-75).

OBJECTIVE: To report on the in vitro and in vivo inhibitory effects of LH-releasing hormone (LH-RH) antagonist Cetrorelix (SB-75; Asta Medica, Frankfurt-Main, Germany) against a panel of human ovarian carcinomas. IN VITRO STUDIES: the effect of SB-75 was measured using a standardized chemosensitivity assay in the following ovarian cancer cell lines: UCI 101; UCI 107; PA-1; NIH: OVCAR 3; UCLA: 222; A2780, parental; A2780-CR, cisplatin resistant; A2780-DR, doxorubicin resistant; and the human breast cancer cell line, MCF-7. Results were expressed as percent growth inhibition determined by crystal violet photometric analysis. In vivo studies: the antiproliferative effect of this agent was examined using UCI-107, a primary epithelial ovarian carcinoma cell line, in a nude mouse model. On day 0, 10 x 10(6) UCI 107 cells were implanted subcutaneously into 20 intact female athymic nude mice (5 to 6 weeks old). On day 8, the mice were randomly divided into two groups of 10; control mice were implanted with miniosmotic pumps filled with a vehicle solution consisting of 5.2% mannitol in saline; and treated animals received pumps filled to deliver continuous administration of SB-75 at 60 micrograms per mouse per day. IN VITRO STUDIES: direct inhibition of cell proliferation by SB-75 was not observed at concentrations ranging from 1 nM to 100 microM (exposure lasting three to four cell doublings) with the exception of MCF-7, which demonstrated a 33% inhibition at the latter concentration. In vivo studies: on day 16, caliper measurements were taken from subcutaneous tumor nodules in SB-75-treated and untreated mice and a significant difference of 270% in mean tumor volume was observed. End point was determined, on day 30, when control tumor volume approached 10,000 mm3. At that time the difference in mean tumor volumes increased to 600%, indicating a substantial antiproliferative effect had been achieved in the SB-75-treated group. CONCLUSION: Our in vitro findings show direct inhibition by SB-75 on proliferation of human breast cancer cells. This direct inhibition in vitro was not observed in our ovarian cancer cell lines. However, in vivo SB-75 caused a significant inhibition of growth of human epithelial ovarian cancer. This may be a result of inhibition of the pituitary gonadal axis and gonadotropin secretion. Our results warrant further investigation.

Toxicity of the mitochondrial poison dequalinium chloride in a murine model system.

Dequalinium chloride (DECA), a cationic, lipophilic mitochondrial poison, selectively targets the mitochondrial membrane of certain epithelial carcinoma cells, in which it inhibits cellular energy production. It has demonstrated potency as a cytotoxic agent specific for carcinomas and may provide a novel approach for cancer therapy, either as a single agent or as an adjunct to conventional chemotherapy. The purpose of this study was to determine the toxicity of DECA in the murine model. One hundred female BALB/c mice were divided into three schedule groups. Group one received a single intraperitoneal (ip) dose of DECA at 10, 15, 20, or 25 mg/kg of body weight. Group two received DECA at 6, 7, 8, 9, or 10 mg/kg ip every other day (QOD), and group three received DECA at 10, 11, 12, 13, or 14 mg/kg ip every 7 days. Over a 30- to 60-day period, acute and subchronic toxicities were evaluated on the basis of the following clinical parameters: respiratory distress, weight loss, and mortality. After a single ip administration, we found a maximum tolerated dose of 15 mg/kg and a lethal dose (LD50) of 18.3 mg/kg. Single ip doses of 20 and 25 mg/kg produced > 50% mortality. Histologic examination of the tissues revealed significant damage to the liver and kidneys, with pulmonary congestion occurring secondary to renal-hepatic failure. A cumulative assessment revealed that 60% of the animals tolerated 15 doses of 6 and 7 mg/kg QOD and that 100% tolerated 5 doses of 11 and 12 mg/kg (every 7 days). Higher DECA doses under either regimen induced severe toxic effects and mortality.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of structural and trace elements in human temporal bone.

This study was undertaken to evaluate a systematic analysis of mineral and trace elements of individual functionally determined parts of adult temporal bone. Marked differences were observed in basic structural elements (Ca, P, Mg, and Zn) among different bone regions. The more so, molar Ca/P ratio was significantly different in various regions, being highest in the hammer and vestibular regions. Taxonomic analysis revealed specific differences in the mineral ratio between the two petrous bone regions believed to develop from various embryonal bases. According to results, the observed differences in mineral trace element composition of particular regions of human temporal bone might be explained by their developmental specificities and functional adaptation.

Holographic analysis of the human pelvis.

Twelve fresh human pelves with preserved lumbar spines, hip joints, and ligaments, were tested by double-exposure and sandwich-hologram interferometry. During physiologic loadings (50-300 N), the pelvis moved as a whole downward and backward. Iliac wings exhibited marked undulation, except for the central part, which showed minor deformations. The sacrum moved downward and rotated forward over an axis 5-9 cm below the promontorium. Removal of the sacroiliac interosseous ligaments eliminated all joint movements and caused a tighter contact between articular surfaces. Removal of the sacrotuberous and sacrospinous ligaments had no influence on the pelvic behavior. The magnitudes of deformations as well as their underlying mineral contents were unequally distributed between the two pelvic sides. These results indicate that the sacroiliac interosseous ligaments are the main determinant of sacral movement. Asymmetric load transmittance to the hip joints might be responsible for the mineral content differences between the pelvic sides.

Trace elements and urinary stone formation: new aspects of the pathological mechanism of urinary stone formation.

The urinary stone, serum and 24-hour urine concentrations of 14 trace elements were determined in urinary stone patients by inductively coupled plasma atomic-emission spectroscopy. The data obtained for 25 active stone patients and 32 whose last stone episode had occurred at least 12 months previously were compared with those of 25 healthy individuals. Urinary nickel, manganese and lithium excretion, and serum nickel, manganese and cadmium concentrations were statistically significantly lower for active stone patients compared to those with previous stone episodes and healthy individuals. No difference in the concentrations of trace elements could be found, however, for patients with previous stone episodes and healthy individuals. Nickel, manganese, lithium and cadmium could be of significance in the pathological mechanism of stone formation, not from mineralogical or crystallographic viewpoints but for the smooth flow of enzymatic reactions in biological systems.

Effects of parathyroidectomy on tissue calcium, phosphorus, magnesium, and copper concentrations in aluminum-loaded uremic rats.

Rats were subjected to a two-stage 5/6 nephrectomy and treated with Al for 2 and 4 wk with a cumulative dose of 4.2 and 8.4 mg of Al, respectively. Other animals were parathyrectomized (PTx) and loaded with 8.4 mg of Al for 4 wk. Total Al, Ca, P, Mg, and Cu contents were analyzed in the liver, kidney, and bone by inductively coupled plasma atomic emission spectrometry (ICP-AES). The results showed that Al given to growing uremic rats significantly increased the content of Al in the liver, kidney, and bone. Moreover, Al treatment increased the liver and kidney Ca levels and decreased the Ca and P values in bone. Previous parathyroidectomy significantly reduced Al accumulation within organs and changes in the Ca and P levels in the bone, liver, and kidney. The result was not influenced by different degrees of renal failure.

The influence of 1 alpha,25- and 24(R),25-dihydroxyvitamin D3 on bone constituents during early mineralization in the rat.

The influence of vitamin D metabolites on intramuscular implants of bone matrix in rachitic rats was investigated. Recipient rats with rickets were injected daily with 1 alpha,25(OH)2D3,24(R),25(OH)2D3 or a combination of both metabolites. The presence of 1 alpha,25(OH)2D3 increased significantly the alkaline phosphatase activity, and slightly increased the activity of acid phosphatase. 24(R),25(OH)2D3 had no effect on the activity of the measured enzymes. The results of inductively coupled plasma emission spectrometric determination of bone elements revealed that: (a) 1 alpha,25(OH)2D3 stimulated the incorporation of magnesium and decreased the phosphorus content of bone implants when compared with rats given both vitamin D metabolites; (b) 1 alpha,25(OH)2D3 as well as 24(R),25(OH)2D3 had antagonistic effects on bone carbonate content. The values for 1 alpha,25(OH)2D3 treated animals were significantly higher, and 24(R),25(OH)2D3 treated rats had a significantly lower carbonate content of implants when compared to the controls. Time-dependent CO2-liberation diagrams indicated a differently bound bone carbonate in 1 alpha,25(OH)2D3 treated rats; (c) when plotted against time, the diagrams for both the values for zinc and the activity distribution of the measured enzymes had a similar appearance, indicating zinc incorporation into bone enzymes during early mineralization. It is concluded that 24(R),25(OH)2D3 should not be compared to 1,25(OH)2D3 on the basis of the same effects, since other effects of 24(R),25(OH)2D3 on the developing bone exist, opposite to those of 1,25(OH)2D3; and these could be important for protecting bone from different agents and in determining the nature of early mineral deposited.

1a,25-Dihydroxyvitamin D3 stimulates alkaline phosphatase activity and inhibits soft-tissue proliferation in implants of bone matrix.

To test the importance of vitamin D metabolites on intramuscular implants of demineralized bone, four-month-old rats were given either 1a,25-(OH)2D3 or 24R,25-(OH)2D3, or a combination of both metabolites, and sacrificed at intervals ranging from five to 35 days after implantation. Histologically there was a reduced ingrowth of mesenchymal cells into the implanted matrix cylinders in the presence of 1a,25-(OH)2D3; the reduction was followed by decreased total DNA and protein values until the 16th experimental day. At 35 days postimplantation, the quantity of new bone was the same in all treated groups. However, 1a,25-(OH)2D3 increased the alkaline phosphatase activity 60%-110% (depending on the denominator used). The metabolite 24R,25-(OH)2R3 had no effect on cell growth or the alkaline phosphatase activity. These results provide evidence for the inhibitory effect of 1a,25-(OH)2D3 on mesenchymal cell growth and its stimulatory effect on osteoblasts, which are responsible for increased alkaline phosphatase activity and new bone formation in vivo.