Significance of histologic pattern of carcinoma and sarcoma components on survival outcomes of uterine carcinosarcoma.

BACKGROUND: To examine the effect of the histology of carcinoma and sarcoma components on survival outcome of uterine carcinosarcoma. PATIENTS AND METHODS: A multicenter retrospective study was conducted to examine uterine carcinosarcoma cases that underwent primary surgical staging. Archived slides were examined and histologic patterns were grouped based on carcinoma (low-grade versus high-grade) and sarcoma (homologous versus heterologous) components, correlating to clinico-pathological demographics and outcomes. RESULTS: Among 1192 cases identified, 906 cases were evaluated for histologic patterns (carcinoma/sarcoma) with high-grade/homologous (40.8%) being the most common type followed by high-grade/heterologous (30.9%), low-grade/homologous (18.0%), and low-grade/heterologous (10.3%). On multivariate analysis, high-grade/heterologous (5-year rate, 34.0%, P = 0.024) and high-grade/homologous (45.8%, P = 0.017) but not low-grade/heterologous (50.6%, P = 0.089) were independently associated with decreased progression-free survival (PFS) compared with low-grade/homologous (60.3%). In addition, older age, residual disease at surgery, large tumor, sarcoma dominance, deep myometrial invasion, lymphovascular space invasion, and advanced-stage disease were independently associated with decreased PFS (all, P < 0.01). Both postoperative chemotherapy (5-year rates, 48.6% versus 39.0%, P < 0.001) and radiotherapy (50.1% versus 44.1%, P = 0.007) were significantly associated with improved PFS in univariate analysis. However, on multivariate analysis, only postoperative chemotherapy remained an independent predictor for improved PFS [hazard ratio (HR) 0.34, 95% confidence interval (CI) 0.27-0.43, P < 0.001]. On univariate analysis, significant treatment benefits for PFS were seen with ifosfamide for low-grade carcinoma (82.0% versus 49.8%, P = 0.001), platinum for high-grade carcinoma (46.9% versus 32.4%, P = 0.034) and homologous sarcoma (53.1% versus 38.2%, P = 0.017), and anthracycline for heterologous sarcoma (66.2% versus 39.3%, P = 0.005). Conversely, platinum, taxane, and anthracycline for low-grade carcinoma, and anthracycline for homologous sarcoma had no effect on PFS compared with non-chemotherapy group (all, P > 0.05). On multivariate analysis, ifosfamide for low-grade/homologous (HR 0.21, 95% CI 0.07-0.63, P = 0.005), platinum for high-grade/homologous (HR 0.36, 95% CI 0.22-0.60, P < 0.001), and anthracycline for high-grade/heterologous (HR 0.30, 95% CI 0.14-0.62, P = 0.001) remained independent predictors for improved PFS. Analyses of 1096 metastatic sites showed that carcinoma components tended to spread lymphatically, while sarcoma components tended to spread loco-regionally (P < 0.001). CONCLUSION: Characterization of histologic pattern provides valuable information in the management of uterine carcinosarcoma.

Factors influencing the persistence of Escherichia coli O157:H7 lineages in feces from cattle fed grain versus grass hay diets.

Escherichia coli O157:H7 is a pathogenic, gram-negative bacterium that causes diarrhea, hemorrhagic colitis, and can lead to fatal hemolytic uremic syndrome in humans. We examined the persistence of E. coli O157:H7 lineages I and II in feces held at 4, 12, and 25 degrees C, from animals fed either grain or hay diets. Three strains of each lineage I and II were inoculated into grain-fed or hay-fed feces, and their persistence was monitored over 28 days. No significant differences in E. coli O157:H7 survival between the 2 lineages in both fecal types was found at the examined temperatures. Volatile fatty acids were higher in grain-fed than in hay-fed feces, resulting in consistently lower pH in the grain-fed feces at 4, 12 and 25 degrees C. Regardless of lineage type, E. coli O157:H7 CFUs were significantly higher in grain-fed than in hay-fed feces at 4 and 25 degrees C. Escherichia coli O157:H7 survival was highest in grain-fed feces at 25 degrees C up to 14 days. Our results indicate that the 2 lineages of E. coli O157:H7 do not differ in their persistence; however, it appears that temperature and feces type both affect the survival of the pathogen.

Escherichia coli O157:H7 strain origin, lineage, and Shiga toxin 2 expression affect colonization of cattle.

Enterohemorrhagic Escherichia coli O157:H7 has evolved into an important human pathogen with cattle as the main reservoir. The recent discovery of E. coli O157:H7-induced pathologies in challenged cattle has suggested that previously discounted bacterial virulence factors may contribute to the colonization of cattle. The objective of the present study was to examine the impact of lineage type, cytotoxin activity, and cytotoxin expression on the amount of E. coli O157:H7 colonization of cattle tissue and cells in vitro. Using selected bovine- and human-origin strains, we determined that lineage type predicted the amount of E. coli O157:H7 strain colonization: lineage I > intermediate lineages > lineage II. All E. coli O157:H7 strain colonization was dose dependent, with threshold colonization at 10(3) to 10(5) CFU and maximum colonization at 10(7) CFU. We also determined that an as-yet-unknown factor of strain origin was the most dominant predictor of the amount of strain colonization in vitro. The amount of E. coli O157:H7 colonization was also influenced by strain cytotoxin activity and the inclusion of cytotoxins from lineage I or intermediate lineage strains increased colonization of a lineage II strain. There was a higher level of expression of the Shiga toxin 1 gene (stx(1)) in human-origin strains than in bovine-origin strains. In addition, lineage I strains expressed higher levels of the Shiga toxin 2 gene (stx(2)). The present study supports a role for strain origin, lineage type, cytotoxin activity, and stx(2) expression in modulating the amount of E. coli O157:H7 colonization of cattle.

Testosterone and social context affect singing behavior but not song control region volumes in adult male songbirds in the fall.

Testosterone (T) induces singing behavior and mediates changes in the sizes and neuroanatomical characteristics of brain regions controlling singing behavior (song control regions, SCRs) in songbirds. These effects may require the enzymatic conversion of T into androgenic and estrogenic metabolites by brain tissues and can be modulated by factors such as season and social context. Testosterone administration to adult male House Finches, Carpodacus mexicanus, in the spring increases the size of their SCRs. Here, we used males of this species to investigate effects of T and T metabolism on brain morphology and singing behavior in the fall. Birds received Silastic capsules containing androgens, estrogens, and/or inhibitors of androgenic action or estrogen synthesis to determine effects of these hormones on song rates and SCR volumes. We also manipulated the social environment by changing the number of birds in visual contact with each other. Testosterone treatment stimulated singing behavior in finches held in small, visually isolated groups and exposed to song playbacks. However, administration of T or T metabolites did not increase SCR sizes. The data suggest that photoperiodic condition and social context may modulate the effects of steroids on SCRs and singing behavior.

Temporal sequence of pulmonary cytokine gene expression in response to endotoxin in C3H/HeN endotoxin-sensitive and C3H/HeJ endotoxin-resistant mice.

Although previous studies suggested that tumor necrosis factor alpha (TNF-alpha) was a critical cytokine responsible for the inflammation observed after exposure to endotoxin, other mediators may also play an important role in the regulation of systemic inflammatory responses independent of TNF-alpha. The present study compared the temporal sequence of endotoxin-induced TNF-alpha, interleukin-1 alpha (IL-1 alpha), and interleukin-10 (IL-10) gene expression and cellular localization of cytokine proteins in pulmonary tissue of two strains of mice that have a genetically based differential sensitivity to endotoxin. Lung tissue and plasma were harvested from endotoxin-sensitive C3H/HeN and endotoxin-resistant C3H/HeJ mice at 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 12 h, and 24 h after intraperitoneal (i.p.) injection of 5 mg/kg endotoxin (Escherichia coli-derived lipopolysaccharide, serotype 0111:B4). There were significant elevations in both TNF-alpha gene and IL-1 alpha expression immediately (15 min) after endotoxin injection in C3H/HeN mice. Although levels of TNF-alpha mRNA in the two mouse strains were similar at 1-2 h, the IL-1 alpha gene expression in pulmonary tissue isolated from endotoxin-resistant mice was not comparable to the levels detected in C3H/HeN endotoxin-sensitive mice at the same times. The most dramatic difference in endotoxin-induced cytokine gene expression between the two strains of mice was in IL-10 mRNA levels in pulmonary tissue isolated from endotoxin-sensitive mice, compared to the lack of detectable increase in IL-10 gene expression in C3H/HeJ endotoxin-resistant mice above baseline at any time point examined. Quantitation of neutrophil infiltration into pulmonary tissue using immunochemical detection of GR-1, a myeloid differentiation-specific antibody, demonstrated that there was a significantly decreased inflammatory infiltrate in pulmonary tissue isolated from C3H/HeJ mice following endotoxin administration, which correlated with decreased levels of proinflammatory cytokine immunoreactive protein within pulmonary cells. Pulmonary cytokine synthesis and immunoreactive protein production did not directly correlate with either the magnitude or the temporal sequence of increases in plasma cytokine levels, suggesting that systemic levels of cytokines may not accurately reflect the cytokine response within the local tissue milieu. The present observations demonstrate that the differential synthesis and production of immunosuppressive cytokines as well as proinflammatory cytokines may be important variables in the determination of the extent of infiltration of inflammatory cells into the local pulmonary site in response to endotoxin and may significantly contribute to the determination of sensitivity or resistance to endotoxin in this murine model.

A comparison of pimozide and haloperidol in the treatment of Gilles de la Tourette's syndrome.

In a double-blind placebo-controlled study, both pimozide and haloperidol significantly decreased tic frequency in nine patients with Gilles de la Tourette's syndrome. These findings are consistent with the catecholamine hypothesis of tic generation. Pimozide was associated with significantly fewer complaints of lethargy. Follow-up 4-20 months later showed that six of seven patients receiving pimozide and one of two receiving haloperidol had had greater than 75% improvement in symptoms.

Production of antibodies to the human thyrotropin receptor and their use in characterising eukaryotically expressed functional receptor.

The structure of the human thyrotropin receptor expressed as a recombinant protein in eukaryotic cells was investigated by immunochemical and functional means using two types of polyclonal rabbit antisera: one raised against the large N-terminal extracellular region (residues 1-415) expressed in E. coli and the other raised against a synthetic peptide (residues 313-330). Both types of antisera gave similar results, with the former being more effective. As expected from the lack of conformation of the immunogens, the antisera worked well in immunoblotting. Less predictably, the antisera also recognised the functional receptor in its native state (detected by flow cytofluorimetry and immunoprecipitation), and inhibited the binding of thyrotropin. Thus the region 313-330 is on the outside of the receptor molecule and falls within, or close to, the binding site of thyrotropin. None of the antisera stimulated cAMP production, showing that this is a very special property, largely restricted to certain human autoantibodies. The antisera were used to immunoprecipitate radioiodinated proteins from Chinese hamster ovary cell (CHO) lines expressing recombinant receptor. The most abundant and reproducible cell-surface molecule that correlated with the presence of full-length functional receptor was a glycopolypeptide of approximately 100 kDa, of which 15 kDa is attributable to carbohydrate, in good agreement with the size predicted for the polypeptide from the cDNA sequence. Three other molecular species were also variably detected at the cell surface: 55 kDa, 180 kDa and large molecular weight material at the top of the polyacrylamide gel.(ABSTRACT TRUNCATED AT 250 WORDS)

In vivo conversion of norethisterone to ethynyloestradiol in perimenopausal women.

The extent to which norethisterone is converted to ethynyloestradiol is controversial. To investigate the conversion of norethisterone to ethynyloestradiol we have used a double isotope infusion technique to measure the conversion in vivo. The use of acids or bases was precluded to prevent possible artefactual formation of phenolic metabolites of norethisterone. Transfer constants for the conversion of norethisterone to ethynyloestradiol in two perimenopausal women were 2.26 and 2.34% as measured in blood and 2.27 and 0.38% in urine. Results from this study show that a small but significant proportion of norethisterone is converted to ethynyloestradiol in vivo.

Topical application of a selective cyclooxygenase inhibitor suppresses UVB mediated cutaneous inflammation.

Ultraviolet B (UVB) radiation causes much of the cutaneous damage after both acute and long-term exposure, and is also the most important etiologic agent in human skin cancer. UVB exposure initially induces an inflammatory response characterized by edema, dermal infiltration of leukocytes, sunburn cell formation, as well as the induction of cyclooxygenase-2 (COX-2) gene expression and subsequent increase in the production and release of prostaglandins. This process of inflammation induced by UVB exposure has been linked to tumor formation. Recently, a specific COX-2 inhibitor, Celecoxib, was developed, which inhibits COX-2-induced inflammation without inhibiting the cytoprotective function of cyclooxygenase-1 (COX-1). The present study compared the effects of topical treatment with Celecoxib (a specific COX-2 inhibitor) and Ibuprofen (a nonspecific COX inhibitor) on the acute UVB-induced cutaneous inflammatory response. We show that the specific inhibition of COX-2 effectively reduced many parameters of UVB-mediated inflammation, including edema, dermal neutrophil infiltration and activation, prostaglandin E2 (PGE2) levels and the formation of sunburn cells. By inhibiting this inflammatory response, topical Celecoxib treatment may ultimately be effective in preventing UVB-induced tumor development in the skin.

Inhibition of cutaneous UV light-induced tumor necrosis factor-alpha protein production by Allotrap 1258, a novel immunomodulatory peptide.

Peptides derived from the heavy chain of the HLA Class-I molecules have been shown to modulate immune responses both in vivo and in vitro. Using a computer-aided rational drug design approach, novel immunomodulatory peptides were designed based on peptide 2702.75-85, derived from HLA-B2702. Several peptides were identified which had increased immunomodulatory activity, including peptides RDP1258 and its D-isomer the peptide Allotrap 1258. The present study using Skh/hr hairless mouse skin model evaluated the in vivo effects of Allotrap 1258 on acute UVB-induced skin inflammation. Here we demonstrate that intraperitoneal administration of Allotrap 1258 1 h prior to UV exposure resulted in significantly diminished levels of UV-induced tumor necrosis factor (TNF)-alpha protein production in the epidermis but had no effect on other parameters of the acute UV-induced inflammatory response. By virtue of its ability to suppress TNF-alpha protein production, Allotrap 1258 could prove to be an effective modulator of inflammatory responses.

The effect of oestrogens and medroxyprogesterone acetate on the uptake of cytotoxic drugs by MCF-7 breast cancer cells.

Previous studies have shown that human breast cancer cells (MCF-7) show an increased response to a number of cytotoxic drugs after 48 h pretreatment with medroxyprogesterone acetate (MPA). As there is evidence that MPA can influence membrane fluidity, we have examined the effect of pre-treatment with MPA on the uptake of methotrexate (MTX) and vincristine (VCR) by MCF-7 cells. The effect of pre-treatment with oestrogen on cytotoxic drug uptake was also examined. After 48 h pre-treatment with MPA (40 or 160 nmol/L), the uptake of MTX was significantly reduced by 14%-44%. Uptake of VCR was also reduced (10%-16%) after pre-treatment of cells with MPA but to a lesser degree than detected for MTX. Pre-treatment with ethinyloestradiol increased the uptake of MTX by up to 45% but enhanced uptake was only detected in cells after exposure to MTX for 1 h. While the results from this study show that oestrogens or MPA can alter the uptake of cytotoxic drugs by MCF-7 breast cancer cells, it is not clear how the MPA dependent decrease in drug uptake enhances the response of MCF-7 to such drugs previously reported.

Comparative ability of ibuprofen and N-(4-hydroxyphenyl)retinamide to inhibit development of rat mammary adenocarcinomas associated with differential inhibition of gene expression of cyclooxygenase isoforms.

A rodent model of carcinogen-induced mammary tumorigenesis was used to determine the comparative growth inhibitory effects of dietary administration of either 1000 mg/kg of the non-steroidal antiinflammatory drug (NSAID) ibuprofen or 1.5 mmol/kg of the synthetic retinoid N-(4-hydroxyphenyl)-retinamide (4-HPR). In addition, the effects of these compounds on gene expression and protein production of the two isoforms of the cyclooxygenase (COX) gene which are responsible for prostaglandin production were examined. Experimental diets were provided to rats beginning at 7 days prior to administration of a single intragastric dose of 15 mg dimethylbenz[a]anthracene (DMBA) and diets were provided ad libitum until the study was terminated at 16 weeks later. Ibuprofen significantly decreased levels of gene expression of both COX-1 and COX-2 (p < 0.01). Although dietary 4-HPR did significantly diminish levels of COX-1 gene expression (p < 0.01) in rat mammary adenocarcinomas, this synthetic retinoid did not significantly inhibit COX-2 gene expression. COX-1 protein was localized to endothelial cells, infiltrating inflammatory cells, and tumor cells, while COX-2 protein was detected primarily within tumor cells. Although ibuprofen was more effective in inhibiting COX-2 gene expression than 4-HPR, ibuprofen and 4-HPR were equally effective in inhibiting development of carcinogen-induced mammary adenocarcinomas.

Inhibition of ultraviolet light B-induced cutaneous inflammation by a specific cyclooxygenase-2 inhibitor.

Ultraviolet B (UVB) radiation is responsible for the majority of cutaneous damage following both acute and long-term exposure, and is believed to be the most important etiologic agent in human skin cancer. UVB carcinogenesis initially induces an inflammatory response characterized by edema, dermal infiltration of leukocytes, as well as the production and release of prostaglandins, which may be critical to the observed damaging effects of UVB light on skin. Recently, a specific cyclooxygenase-2 (COX-2) inhibitor, Celecoxib, was developed, which inhibits COX-2-induced inflammation without inhibiting the cytoprotective function of cyclooxygenase-1 (COX-1). Studies have demonstrated that oral administration of Celecoxib decreased the incidence of skin and colon tumors. Recently, the process of inflammation has been linked to tumor formation. The present study examined the effects of a topical application of Celecoxib on the acute UVB-induced cutaneous inflammatory response. We show that topical Celecoxib treatment effectively reduced many parameters of UVB-mediated inflammation, including edema, dermal myeloperoxidase activity, neutrophil infiltration, and prostaglandin E2 (PGE2) levels. By inhibiting this inflammatory response, topical Celecoxib treatment could ultimately be effective in preventing tumor development and progression in the skin, which is known to result from long-term UV exposure.

Posterior tibial tendon dysfunction: secondary MR signs.

We evaluated four potential secondary magnetic resonance imaging signs to aid in clinical diagnosis of posterior tibial tendon (PTT) tears. Seventy-one ankles (25 PTT tears and 46 controls) were evaluated for the following secondary signs: (1) PTT sheath fluid, (2) a distal tibial spur located just anterior to the PTT, (3) unroofing of the talus, and (4) "bone bruise"--like medullary lesions. Two musculoskeletal radiologists rated their confidence using a scale and were compared for level of agreement. The presence of PTT sheath fluid had modest specificity and fair to moderate sensitivity. Tibial spurring and unroofing of the talus had excellent specificity and fair sensitivity. Bone bruise-like lesions were commonly seen in cases and controls. Examination of divergence of opinion between the two radiologists revealed pitfalls in interpretation of PTT sheath fluid and bone bruise-like lesions, which were commonly the result of adjacent vessels and inhomogeneous fat saturation, respectively. We conclude that secondary signs of PTT tears with high specificities include unroofing of the talus, tibial spurring, and PTT sheath fluid.

Quality assurance of histopathologic diagnosis in the British Army: role of the Army Histopathology Registry in completed case review.

The purpose of this study was to assess the clinical and pathological value of reports resulting from review of all completed surgical pathology cases submitted to the Army Histopathology Registry (AHR). All histopathological cases completed in the British Army are sent to the AHR for archiving; prior to placing cases in the archive both microscopic material and submitted reports are reviewed by staff of the AHR. A "nonagreed" report is produced for those cases in which the reviewing pathologist has a dissenting opinion or for which he thinks other comments may be helpful. All nonagreed reports produced over a 19 month period were subjected to a further pathological and clinical review. The original surgical pathology reports were compared with AHR reports and the significance of the differences in diagnosis assessed. During the study interval, 4.0% of total cases reviewed were identified as nonagreed record cases. The clinical and pathological reviews placed the nonagreed cases into significant categories in 2.1% and 1.9% of instances respectively. These findings suggest that nonselected review of completed surgical pathology cases identifies a significant proportion of cases for which dissenting opinions may have important clinical and pathological consequences.

Potential for in situ chemical oxidation of acid extractable organics in oil sands process affected groundwater.

The process of bitumen extraction from oil sands in Alberta, Canada leads to an accumulation of toxic acid-extractable organics (AEOs) in oil sands process water (OSPW). Infiltration of OSPW from tailings ponds and from their retaining sand dykes and subsequent transport towards surface water has occurred. Given the apparent lack of significant natural attenuation of AEOs in groundwater, remediation may be required. This laboratory study evaluates the potential use of unactivated persulfate and permanganate as in situ oxidation agents for remediation of AEOs in groundwater. Naphthenic acids (NAs; CnH2n+zO2), which are a component of the acutely toxic AEOs, were degraded by both oxidants in OSPW samples. Permanganate oxidation yielded some residual dissolved organic carbon (DOC) whereas persulfate mineralized the AEO compounds with less residual DOC. Acid-extractable organics from oxidized OSPW had essentially no Microtox toxicity.

The spectral properties of (-)-epigallocatechin 3-O-gallate (EGCG) fluorescence in different solvents: dependence on solvent polarity.

(-)-Epigallocatechin 3-O-gallate (EGCG) a molecule found in green tea and known for a plethora of bioactive properties is an inhibitor of heat shock protein 90 (HSP90), a protein of interest as a target for cancer and neuroprotection. Determination of the spectral properties of EGCG fluorescence in environments similar to those of binding sites found in proteins provides an important tool to directly study protein-EGCG interactions. The goal of this study is to examine the spectral properties of EGCG fluorescence in an aqueous buffer (AB) at pH=7.0, acetonitrile (AN) (a polar aprotic solvent), dimethylsulfoxide (DMSO) (a polar aprotic solvent), and ethanol (EtOH) (a polar protic solvent). We demonstrate that EGCG is a highly fluorescent molecule when excited at approximately 275 nm with emission maxima between 350 and 400 nm depending on solvent. Another smaller excitation peak was found when EGCG is excited at approximately 235 nm with maximum emission between 340 and 400 nm. We found that the fluorescence intensity (FI) of EGCG in AB at pH=7.0 is significantly quenched, and that it is about 85 times higher in an aprotic solvent DMSO. The Stokes shifts of EGCG fluorescence were determined by solvent polarity. In addition, while the emission maxima of EGCG fluorescence in AB, DMSO, and EtOH follow the Lippert-Mataga equation, its fluorescence in AN points to non-specific solvent effects on EGCG fluorescence. We conclude that significant solvent-dependent changes in both fluorescence intensity and fluorescence emission shifts can be effectively used to distinguish EGCG in aqueous solutions from EGCG in environments of different polarity, and, thus, can be used to study specific EGCG binding to protein binding sites where the environment is often different from aqueous in terms of polarity.

Metabolism of beclamide, N-benzyl-3-chloropropionamide, in man.

1. Following oral administration of [14C]beclamide to human subjects there was almost complete elimination of radioactivity in the urine within 48 h. 2. Less than 1% of the administered dose was excreted unchanged. 3. The major metabolites were (a) N-(4-hydroxybenzyl)-3-chloropropionamide, (4-hydroxybeclamide; c. 56%), which was eliminated to a small extent in the free form and as a sulphate but mainly as a glucuronide, and (b) hippuric acid (c. 20%). 4. Six minor metabolites were also detected, of which the following were identified: benzoic acid (up to c. 7%), N-(4-hydroxybenzyl-3-chloro-2-hydroxypropionamide, (dihydroxy-beclamide; c. 2%, mostly as a glucuronide) and 4-hydroxybenzoic acid (c. 0.6%, partly as a glucuronide).