Correlation of expression of cyclooxygenase-2, vascular endothelial growth factor, and peroxisome proliferator-activated receptor delta with head and neck squamous cell carcinoma.

Cyclooxygenase (COX) is the rate-limiting enzyme in the formation of prostaglandins from arachidonic acid. COX exists in 2 isoforms, COX-1 and COX-2. These isoforms are encoded by separate genes and demonstrate cell-specific expression and regulation. Peroxisome proliferator-activated receptor delta (PPARdelta) is a nuclear transcription factor that is activated by prostacyclin. Vascular endothelial growth factor (VEGF) is a proangiogenic factor that is up-regulated in various tumors. Vascular endothelial growth factor has been shown to interact with COX-derived prostaglandins in angiogenesis. To better understand the roles of these genes in head and neck squamous cell carcinoma (HNSCCA), we examined the differential expression of the COX1, COX2, VEGF, and PPARdelta genes in these tumors. Tissue samples from patients with HNSCCA were analyzed for COX-1, COX-2, VEGF, and PPARdelta messenger RNAs (mRNAs) by in situ hybridization. COX-1 and COX-2 mRNAs were also evaluated with Northern blot hybridization. Immunohistochemistry was used to analyze for COX-2 and PPARdelta proteins. Results showed focal areas of accumulation for COX-2, VEGF, and PPARdelta but not COX-1 in human HNSCCA. Northern blot hybridization showed higher levels of COX-2 mRNA in HNSCCA than in normal tissue. This suggests a supportive role of COX-2 in development and/or progression of HNSCCA. In addition, PPARdelta may be a receptor for COX-2-produced prostaglandins in HNSCCA. There is a potential role for selective COX-2 inhibitors in the treatment of these lesions.

Characterization of cyclooxygenase in laryngeal papilloma by molecular techniques.

OBJECTIVES: Demonstrate the induction of cyclooxygenase-2 (COX-2) in laryngeal papilloma Discuss the possible causal role of COX-2 in papilloma formation. Consider the potential for treatment of papilloma using selective COX-2 inhibitors. STUDY DESIGN: Molecular biological analysis of COX-1 and COX-2 in laryngeal papilloma. METHODS: Tissue samples from five patients with recurrent respiratory papillomatosis (RRP) were analyzed by in situ hybridization, immunohistochemical staining, and reverse transcription polymerase chain reaction (RT-PCR) techniques. RESULTS: In situ hybridization to COX-2 mRNA showed strong autoradiographic signal surrounding fibrovascular cores. COX-1 autoradiographic signal was low intensity or nondetectable. Normal buccal mucosa biopsies showed low-density or nondetectable autoradiographic signal for both COX-1 and COX-2 mRNAs. In situ hybridization results were corroborated by RT-PCR studies. Levels of COX-2 mRNA were 13-fold more than those in normal mucosa. Immunohistochemical staining for COX-1 and COX-2 showed a similar pattern to that seen with in situ hybridization in both normal and papilloma tissues. CONCLUSIONS: There is an elevation of COX-2 expression in papilloma tissues. This may represent a causal role of COX-2 in the formation and proliferation of laryngeal papilloma. There may also be a role for selective COX-2 inhibition for the treatment of

Expression of interleukin 1, interleukin 6 and tumour necrosis factor alpha in mouse uterus during the peri-implantation period of pregnancy.

Interleukin 1 (IL-1), IL-6 and tumour necrosis factor alpha (TNF-alpha) are expressed in the mouse uterus on days 1-3 of pregnancy. Cytokine production is temporally associated with the post-mating intrauterine acute inflammatory response. In this study, IL-1, IL-6 and TNF-alpha were detected in the uterus of pregnant mice from day 3 to day 9, using northern blotting, bioassays and immunocytochemistry. IL-1 bioactivity increased from a low concentration on day 3 to a peak between days 4 and 5 and decreased to low concentrations on days 7 and 8. Blastocyst implantation occurs late on day 4. IL-6 bioactivity was high from day 3 to day 9 and activity was maximal on days 5 and 6. TNF-alpha bioactivity increased from its lowest concentration on day 3 to a peak on day 8. Although changes in bioactivity concentrations occurred at different times from changes in mRNA concentrations, the changes were approximately parallel. Translation of mRNA into an immunologically detectable product was confirmed using immunocytochemistry with polyclonal anti-cytokine antibodies. We conclude that the cytokines IL-1, IL-6 and TNF-alpha are produced in the uterus during the peri-implantation period of pregnancy in mice. Changes in cytokine concentrations suggested the existence of some form of regulated expression.

Interleukin-1, interleukin-6, and tumor necrosis factor alpha are produced in the mouse uterus during the estrous cycle and are induced by estrogen and progesterone.

The ovarian steroids, estrogen and progesterone, regulate cellular and molecular changes which occur in the uterus during the estrous cycle. Cycles of protein synthesis, cell proliferation and differentiation, and cell death are the direct results of changes in hormone concentration. To explore the possibility that cytokines, which stimulate proliferation and differentiation of numerous types of cells, might be associated with those cyclic changes, the production of IL-1, IL-6, and TNF alpha was examined in the mouse uterus. Cytokine mRNA expression, bioactivity, and immunoreactivity were quantitated during the estrous cycle, following ovariectomy and exposure of ovariectomized mice to estrogen and progesterone. IL-1, IL-6, and TNF alpha mRNA was detected, and mRNA levels for each of the cytokines varied with the stage of the cycle. Cytokine bioactivity was expressed throughout the cycle, but levels of each cytokine were highest during proestrus and/or estrus. Immunoreactivity paralleled bioactivity. Uterus from ovariectomized mice contained little or no cytokine activity, and systemic administration of estrogen or progesterone resulted in the induction of IL-1 alpha and IL-1 beta mRNA expression. Significant amounts of IL-6 and TNF alpha mRNA appeared only following the exposure of ovariectomized mice to estrogen plus progesterone. Cytokine bioactivity and immunoreactivity also appeared following the administration of estrogen and/or progesterone. The highest activity levels for each cytokine were observed following the injection of estrogen plus progesterone. Cyclic expression of IL-1, IL-6, and TNF alpha in the uterus and their apparent regulation by estrogen and progesterone raise the possibility that cytokines and factors which are induced by cytokines are part of the regulatory process which is induced by ovarian hormones in the uterus of reproductive age females.

Expression of colony-stimulating factors and inflammatory cytokines in the uterus of CD1 mice during days 1 to 3 of pregnancy.

Morphological and immunohistochemical analyses have documented the development of an acute inflammatory response, marked by the early appearance of granulocytes and later infiltration of mononuclear cells, in the uterus immediately after mating in mice. The response peaked on Day 1 and subsided by Day 3. In the present study, RNAs for macrophage colony-stimulating factor (CSF-1) and granulocyte-macrophage colony-stimulating factor (GM-CSF) and for interleukin 1 alpha (IL-1 alpha), IL-1 beta, interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF-alpha) were detected in uterine tissue on Day 1. With the exception of IL-6, which was higher on Day 3 than on Day 1, and IL-1 alpha, which was not reduced on Day 2, concentrations of cytokine mRNA decreased to Day 3. No bioactivity was detected for GM-CSF, granulocyte colony-stimulating factor or IL-3, but CSF-1, IL-1, IL-6 and TNF-alpha were detected on Day 1 using bioassays. Changes in concentrations approximately paralleled those for mRNA. The concentrations of mRNA for CSF-1, IL-1, IL-6 and TNF-alpha were higher on Day 1 of pregnancy than in the uteri of cycling mice 24 h earlier. The data are consistent with previous morphological observations demonstrating the expression of an acute inflammatory response in the mouse uterus after mating. Further, the data demonstrate the expression of genes for CSF-1, GM-CSF, IL-1 alpha, IL-1 beta, IL-6 and TNF-alpha is induced in the uterus during mating.

Acute poisoning with a glyphosate-surfactant herbicide ('Roundup'): a review of 93 cases.

Between 1 January 1980, and 30 September 1989, 93 cases of exposure to herbicides containing glyphosphate and surfactant ('Roundup') were treated at Changhua Christian Hospital. The average amount of the 41% solution of glyphosate herbicide ingested by non-survivors was 184 +/- 70 ml (range 85-200 ml), but much larger amounts (500 ml) were reported to have been ingested by some patients and only resulted in mild to moderate symptomatology. Accidental exposure was asymptomatic after dermal contact with spray (six cases), while mild oral discomfort occurred after accidental ingestion (13 cases). Intentional ingestion (80 cases) resulted in erosion of the gastrointestinal tract (66%), seen as sore throat (43%), dysphagia (31%), and gastrointestinal haemorrhage (8%). Other organs were affected less often (non-specific leucocytosis 65%, lung 23%, liver 19%, cardiovascular 18%, kidney 14%, and CNS 12%). There were seven deaths, all of which occurred within hours of ingestion, two before the patient arrived at the hospital. Deaths following ingestion of 'Roundup' alone were due to a syndrome that involved hypotension, unresponsive to intravenous fluids or vasopressor drugs, and sometimes pulmonary oedema, in the presence of normal central venous pressure.