Expression of aromatase cytochrome P450 protein and messenger ribonucleic acid in human endometriotic and adenomyotic tissues but not in normal endometrium.

To determine whether local estrogen production takes place in endometriotic or adenomyotic tissues, in eutopic endometrium from patients with endometriosis or adenomyosis, and in normal endometrium, tissue specimens were examined by immunohistochemistry, catalytic activity, and mRNA expression for aromatase cytochrome P450 (P450arom). P450arom was immunohistochemically localized in the cytoplasm of glandular cells of endometriotic and adenomyotic tissues, and of eutopic endometrium from patients with the respective diseases, whereas estrogen receptors and progesterone receptors were localized in the nuclei of the glandular cells and stroma. Aromatase activity in the microsomal fraction of adenomyotic tissues was inhibited by the addition of danazol, aromatase inhibitors, and anti-human placental P450arom monoclonal antibody (mAb3-2C2) in a manner similar to such inhibition in other human tissues. Reverse transcription polymerase chain reaction and Southern blot analysis also revealed P450arom mRNA in these tissues. However, neither P450arom protein activity nor mRNA was detected in endometrial specimens obtained from normal menstruating women with cervical carcinoma in situ but without any other gynecological disease. These results suggest that at a local level, endometriotic and adenomyotic tissues produce estrogens, which may be involved in the tissue growth through interacting with the estrogen receptor.

Endoplasmic reticulum proliferates without an increase in cytochrome P-450 in hepatocytes of mice treated with phenobarbital and cobalt chloride.

To determine whether endoplasmic reticulum (ER) proliferation in hepatocytes after phenobarbital (PB) administration relates closely to cytochrome P-450 (P-450) increase, we have measured the amount of total P-450 per unit cytoplasmic volume (P-450 content) by microphotometry and estimated the area of ER per unit cytoplasmic volume (ER area) by morphometry in periportal, midzonal, and perivenular hepatocytes of mice injected daily with PB (100 mg/kg), or with PB (100 mg/kg) plus cobalt chloride (50 mg/kg) for three days. After injection of PB, the P-450 content and ER area increased in hepatocytes of the three sublobular zones. In mice treated with PB plus cobalt chloride, however, the ER area increased, but the P-450 content decreased or remained unchanged in hepatocytes of the three zones. We conclude that cobalt chloride inhibits the increase in total P-450 but has no effect on the proliferation of ER of hepatocytes in mice treated with PB, indicating a dissociation of ER proliferation and P-450 increase after administration of PB.

Absence of paravascular nerve projection and cross-innervation in interscapular brown adipose tissues of mice.

To determine whether a paravascular nerve projection or cross-innervation exists in the interscapular brown adipose tissue (BAT), the distribution of noradrenergic or peptidergic nerve fibers in intact or denervated interscapular BAT pads of mice was examined histochemically. Noradrenaline (NA) fibers were visualized by the glyoxylic acid condensation method, and neuropeptide Y (NPY) and substance P (SP) fibers were detected immunohistochemically. Numerous NA-positive fibers and a few NPY- or SP-positive fibers were observed around intralobular arterioles in intact BAT pads. Some NA-positive fibers and very few NPY- or SP-positive fibers were seen around interlobular arteries. NA- and NPY-positive fibers were also found around brown adipose cells in the parenchyme of BAT, whereas SP-positive fibers were absent around the cells. However, all sections cut from denervated BAT pads of unilaterally or bilaterally denervated animals showed a total absence of NA-, NPY- or SP-positive nerve fibers. Therefore, neither a paravascular projection of NA, NPY, and SP fibers to the BAT nor a cross-innervation of these nerve fibers between the left and right BAT pads exists in the mouse BAT.

Significance of high glucose-6-phosphatase activity in rat oviduct epithelium.

To study the origin of glucose in the oviduct fluid, we cytochemically examined glucose-6-phosphatase (G6Pase) activity in rat oviduct. The activity in the whole oviduct was also assayed biochemically. During proestrous, estrous, and metestrous phases, staining reaction for the activity was moderate in the epithelium of the caudal isthmus (CaI) and uterotubal junction (UJ), whereas it was weak in that of the ampulla (A) and cephalic isthmus (CeI). In the diestrous phase, staining reaction in the epithelium of CaI and UJ became strong although it remained weak in that of A and CeI. Reaction product for the activity was localized in the endoplasmic reticulum and nuclear envelope of all cell types in the epithelium. The amount of reaction product in secretory cells was small to moderate in CaI and UJ, and small in A and CeI during proestrus, estrus, and metestrus. In diestrous the amount became abundant in CaI and UJ and moderate in A and CeI. However, the amount in ciliated cells remained small in the four segments during the four phases. The biochemical activity in diestrous was greater than that in proestrus, estrus, or metestrus. This shows that the activity is high in secretory cells in the epithelium of CaI and UJ in the diestrous phase and suggests that the role of the high activity is to release glucose into the oviduct fluid for use by the embryo passing down the CaI and UJ to the uterus.

Clinical study of a new fecal occult blood test using a combination assay of hemoglobin and transferrin.

The applicability of a new immunological fecal occult blood test in which hemoglobin (Hb) and transferrin (Tf) are simultaneously assayed was evaluated. The mean absorbance and standard deviation (510/630 nm) obtained by this test was 0.840 +/- 0.805 in 51 fecal samples from patients with colon cancer, 0.248 +/- 0.305 in 95 samples from patients with colon polyps, and 0.104 +/- 0.053 in 110 samples from control patients; these values differed significantly (P less than 0.005). Hb and Tf concentrations were separately determined in the same fecal samples, and qualitative evaluation was performed with a cutoff value of 5.1 micrograms/g feces for Hb and 0.4 micrograms/g feces for Tf. Hb or Tf was positive in 41 of the 51 samples in the colon cancer group, 33 of the 95 in the colon polyp group, and 3 of the 110 in the control group. Qualitative analysis of the values obtained by the combination assay of Hb and Tf with a cutoff value of 0.200 revealed positive rates of 41/51 in the colon cancer group, 33/95 in the colon polyp group, and 4/110 in the control group. These results suggest the usefulness of a combination assay of Hb and Tf as a fecal occult blood test.