Estrogen and progesterone receptors, cell proliferation, and c-fos expression in the ovine uterus during early pregnancy

To evaluate uterine growth during pregnancy and the potential roles of estrogen and progesterone in regulating uterine cell proliferation and c-fos expression, ewes were assigned randomly to slaughter on day 12 after estrus (nonpregnant, NP), and on days 12, 18, 24, or 30 after mating (pregnant, P) in Exp 1 (n = 7 ewes/day) and on days 12 or 14 after estrus and on days 12, 14, 16, 18, 21, or 24 after mating in Exp 2 (n = 3-6 ewes/day). In Exp 1, endometrial expression of c-fos mRNA was evaluated, and labeling index was determined both in vitro (incorporation of 3H-thymidine) and in vivo (iv injection of bromodeoyxuridine [BrdU], a thymidine analog). Endometrial expression of the c-fos proto-oncogene was increased by approximately 10-fold on days 18, 24, and 30 P compared with day 12 NP or P. Labeling index (proportion of cells incorporating 3H-thymidine or BrdU, which provides an index of the rate of cell proliferation) of endometrial caruncular and intercaruncular tissues was low for day 12 NP or P, increased on day 18 P, and remained elevated on days 24 P and 30 P. On day 18 P, labeling index also was greater for gravid than nongravid horns for both caruncular and intercaruncular tissues. In Exp 2, estrogen receptors (ER), progesterone receptors (PR), and proliferating (BrdU-positive) cells were immunolocalized. The percentage of cells exhibiting specific staining for ER, PR, and BrdU was quantified morphometrically for epithelial, stromal, and glandular tissues within luminal and deep regions, as well as for myometrial tissues. For luminal epithelium and glands, the rate of cell proliferation increased dramatically by day 18 P, even though ER and PR levels were low in these compartments. Conversely, the rate of cell proliferation remained low throughout early pregnancy in deep glands, deep stroma, and myometrium, in association with sustained or transient increases in ER and PR levels. For luminal stroma, the rate of cell proliferation increased by day 21 P even though ER levels were low and PR levels remained high. Thus, during early pregnancy, c-fos expression increased concomitantly with increased endometrial cell proliferation. In addition, during early pregnancy, ER and PR levels were inversely related to the rate of cell proliferation in most of the uterine tissue compartments except luminal stroma, which exhibited increased cell proliferation even though ER levels were low and PR levels remained high.     

Estrogen and progesterone receptors in endometrial carcinoma

Malignant rhabdoid tumor (MRT) is a recently described variety of childhood renal neoplasm. MRT arising primarily in the central nervous system (CNS) is still a rather unfamiliar pathological entity and is frequently misdiagnosed as medulloblastoma or primitive neuroectodermal tumor (PNET). We describe a 7-month-old boy who harbored a CNS-MRT that originated within the IV ventricle and invaded the brain stem and the cerebellar hemispheres. After an initial documented good response to chemotherapy the tumor recurred locally 9 months after operation. We discuss clinical and histopathological features distinguishing between PNET/medulloblastoma and MRTs. Current literature on MRT of the CNS is briefly reviewed.     

Immunohistochemical localization of somatostatin receptors sst2A in human tumors

Human tumors frequently express somatostatin receptors. However, none of the receptor subtype proteins have been individually visualized in normal or neoplastic human tissues. Here, the distribution of the sst2A receptor was investigated using immunohistochemistry with the specific anti-peptide antibody R2-88 in 47 human tumors. All tumors selected for their abundance of sst2 mRNA and/or strong binding of the sst2-preferring ligand 125I-labeled Tyr3-octreotide were specifically immunostained with R2-88. Conversely, all tumors without somatostatin binding or expressing predominantly other somatostatin receptor subtype mRNAs (sst1 or sst3) were not specifically immunostained by R2-88. Specificity was shown in immunoblots, demonstrating receptor migration as a 70-kd broad band. In immunohistochemical and immunoblotting experiments, the abolition of staining after antibody blockade with antigen peptide was demonstrated. Immunostaining was identified in cryostat and in formalin-fixed, paraffin-embedded sections. Heat-induced epitope retrieval was necessary to visualize sst2A receptors in formalin-fixed sections. Moreover, because of occasional high nonspecific staining, the demonstration of complete abolition of immunostaining by treatment with antigen peptide was a prerequisite for the correct identification of sst2A-positive tumors. The sst2A receptors were clearly located at the membrane of the tumor cells. These results provide the first localization of a somatostatin receptor subtype in human tissues at the cellular level. The sst2A receptor identification and visualization in tumors with simple immunohistochemical methods in formalin-fixed, paraffin-embedded material will open new diagnostic opportunities for pathologists.     

Immunohistochemical localization of glucose transporters and insulin receptors in human fetal membranes at term

The localization has been investigated of the isoforms GLUT1, GLUT3 and GLUT4 of glucose transporter proteins as well as of insulin receptors. Fetal membranes (n = 10) were examined by immunohistochemical methods at the light and electron microscopic levels using mono- and polyclonal antibodies. In all amnion epithelial cells, GLUT1 and GLUT3 antibodies were bound to the apical membrane. Very rarely the GLUT1 antibody also immunostained the basolateral membrane and reacted weakly with the endomembrane system and membranes of the lateral cell protrusions. Fibroblasts reacted with the antibodies against GLUT1, GLUT4 and insulin receptor, whereas they were labelled only in one case with GLUT3 antibody. Cytotrophoblast cells were only stained with antibodies against GLUT1 and GLUT3. Antibodies against GLUT4 only reacted with fibroblasts in the membranes. On amnion epithelial cells, weak immunoreactivity with insulin receptor antibodies was detected only at the electron microscopic level. The data indicate: (1) GLUT1 is located on all cells of the amnion, whereas GLUT3 is present in detectable amounts only on amnion epithelial cells and cytotrophoblast; (2) GLUT1 and GLUT3 on amnion epithelial cells are predominantly located on the apical surface; (3) GLUT4 and insulin receptors are not regularly expressed. We suggest that amnion epithelial cells cover their basal glucose requirements from the amniotic fluid and not from the maternal circulation.     

Immunohistochemical analysis of distribution of estrogen receptors and progesterone receptors in the postmenopausal endometrium

OBJECTIVE: To examine the expression of sex steroid receptors (ER: estrogen receptor; PR: progesterone receptor) in the postmenopausal endometrium (PMEM) and the relationship to clinical data for studying its characters. METHODS: The immunohistochemical reactivity of the PMEM was studied using monoclonal antibodies against ER and PR, in 33 postmenopausal patients. RESULTS: The endometrium was thicker in patients who were postmenopausal for 1 to 10 years (1.48 +/- 1.31 mm) than in patients who were postmenopausal for more than 10 years (0.79 +/- 0.37 mm)(p < 0.05). Among the 33 postmenopausal endometrial samples, ER positivity was found in the glands in 26 cases (78.8%) and PR positivity was detected in 18 cases (54.5%). The average age of the patients with ER positive reactivity in the glands (61.69 +/- 7.26 years) was significantly lower than that of the patients with ER negative reactivity (66.00 +/- 3.56 years)(p < 0.05). Furthermore, the endometrial thickness of the patients with ER or PR positive reactivity in the glands (1.24 +/- 1.09 mm and 1.47 +/- 1.20 mm, respectively) was significantly greater than that of the patients with ER or PR negative reactivity (0.67 +/- 0.26 mm and 0.70 +/- 0.40 mm, respectively)(p < 0.05). CONCLUSION: ER in the glands of the PMEM was determined to decrease gradually with increased aging. The presence of ER and PR in the gland cells seemed likely to determine the thickness of the PMEM.     

Immunohistochemical localization of endothelin-1 in placenta and fetal membranes in term and preterm human pregnancy

A case is reported in which an intruded incisor was initially treated by an endodontic dressing with calcium hydroxide and then extruded using a removable orthodontic appliance. A follow-up examination seven years after completion of endodontic therapy and bleaching showed a favourable response. A review of the relevant literature indicates that intrusion occurs in five to twelve per cent of luxation cases. In this type of injury maximum damage occurs to the pulp and all supporting structures because the tooth is driven into the alveolar process. Complications which have been reported include: pulp necrosis, apical radiolucencies, partial or total pulp calcification, root resorption (surface, inflammatory or replacement), marginal periodontal bone breakdown, and arrested or disturbed root development. The prognosis for pulp survival after intrusion is much more favourable for teeth with incomplete root formation than for teeth with complete root formation. Treatment options available to bring an intruded tooth into alignment are: to await spontaneous re-eruption which may occur if root formation is incomplete, uncovering of the intruded crown, orthodontic extrusion which is allied with gentle luxation if the tooth does not move, and immediate surgical repositioning.     

Estrogen and progesterone receptors in endometrial carcinoma: comparison of immunohistochemical and biochemical analysis

In 159 endometrial carcinomas, estrogen (ER) and progesterone receptors (PR) were determined biochemically by dextran-coated charcoal (DCC) assay and immunohistochemically (ICA) on frozen sections. ICA receptor content was estimated by a total histologic score (HSCORE), including all tissue components, and by a cancer HSCORE, including malignant cells only. These scores were closely correlated. A single biopsy was found to be representative for each tumor. ER-DCC status was positive in 90.3% and PR-DCC status in 92.2% of the tumors. ER total HSCORE was positive in 47% and PR total HSCORE in 89% of tumors. ER and PR correlated inversely with tumor grade (p < 0.001). Correlations were found between ER and PR content determined by either method (DCC: r = 0.77; ICA: r = 0.50), as well as between DCC and ICA content (ER: r = 0.52; PR: r = 0.76). The association between DCC and ICA was affected by the tumor grade: the DCC values decreased relatively more than total HSCOREs with increasing grade. The sensitivity of ICA against DCC assay was 56% for ER and 86% for PR. Maximal agreement between receptor status as determined by ICA and by DCC would result from a DCC cutoff level of 130 fmol/mg for ER and 114 fmol/mg for PR.     

Immunohistochemical detection of estrogen and progesterone receptors in spermatozoa of infertile men

OBJECTIVE: To evaluate the clinical implication of sex steroidal actions on human spermatozoa. PATIENTS AND METHODS: Human spermatozoa were obtained from 20 males. Immunohistochemical staining for estrogen receptors (ER) and progesterone receptors (PR) was conducted by the avidin-biotin-peroxidase complex method. RESULTS: Positive immunohistochemical staining of ER and PR was observed in all 8 cases of fertile couples. From infertile couples, ER were detected in 8/12 cases, including 3 with normozoospermia, 2 with oligozoospermia, 1 with asthenozoospermia, and 2 with teratozoospermia, but not in 4/12 cases, 1 with normozoospermia, 1 with asthenozoospermia, and 2 with oligoasthenozoospermia. PR were undetectable in all cases of negative ER and in 1 case (normozoospermial) of positive ER. CONCLUSION: Negative ER and/or PR in spermatozoa from infertile couples might be involved in cases of male infertility.     

Immunohistochemical localization of endothelin-1, endothelin-3 and endothelin receptors in human pheochromocytoma and paraganglioma

Endothelin (ET) and its receptor system have been shown to exert various biological effects on different types of cells in addition to their well-known vasoconstrictor activity. Recently ET-1, ET-3 and the ETB receptor have been shown to play an important role in the development of neural crest-derived cells and, in this context, pheochromocytomas have been reported to harbor ET-1. Endothelin-3 or ET receptor subtypes, however, have not been examined in pheochromocytoma and paraganglioma so far. In the present study the immunohistochemical localization of ET-1/big ET-1, ET-3/big ET-3 and the ETA and ETB receptors were investigated to clarify the biological characteristics of these two tumors using 32 pheochromocytomas and 11 extra-adrenal paragangliomas. Endothelin-1/big ET-1 was detected in 19 pheochromocytomas (59%) and eight paragangliomas (72%), while ET-3/big ET-3 was detected in 10 pheochromocytomas (31%) and three paragangliomas (27%). The ETA receptor was found in 21 pheochromocytomas (66%) and in eight paragangliomas (73%), while the ETB receptor was found in 25 pheochromocytomas (78%) and in eight paragangliomas (73%). Normal adrenomedullary cells lacked each antigen examined. Endothelin-immunoreactive tumor cells were distributed focally or in a manner scattered, while receptor-immunostained tumor cells were distributed with a focal pattern for the ETA receptor and with a focal or diffuse pattern for the ETB receptor. Endothelin and its receptor coexisted in the same tumor in 21 of 28 ET-positive pheochromocytomas and in eight of 10 ET-positive paragangliomas. In addition, seven pheochromocytomas and two paragangliomas revealed positivity of the receptor(s) irrespective of the absence of ET-immunoreactivity. In conclusion, ET and its receptor are frequently and concomitantly expressed in the pheochromocytoma and paraganglioma. From the highly frequent expression of this system or the receptor(s), ET-receptor-mediated signal transduction of these tumors concerning growth and/or cell survival is expected, although definite biological significance of this ligand-receptor system in these tumors awaits further investigation.     

Localization and expression of tissue kallikrein and kallistatin in human blood vessels

Human lysosomal acid lipase/cholesteryl ester hydrolase (hLAL) is essential for the intralysosomal metabolism of cholesteryl esters and triglycerides taken up by receptor-mediated endocytosis of lipoprotein particles. The key role of the enzyme in intracellular lipid homeostasis is illustrated by two lysosomal storage diseases inherited as autosomal recessive traits. Wolman disease, associated with deficient hLAL activity, leads to massive intracellular substrate accumulation and is always fatal in early infancy. Cholesteryl ester storage disease (CESD), in contrast, is characterized by very low levels of enzymic activity sufficient to allow survival of the affected patients into adulthood. In order to elucidate the underlying molecular defects in Wolman disease, we have characterized the hLAL gene in two female Wolman patients of German and Turkish origin by SSCP and DNA sequence analysis. Our results demonstrate that the German proband was compound heterozygous for an 8-bp deletion in exon 3 and a 2-bp deletion in exon 4 of the hLAL gene. These frameshift mutations lead to protein truncation at amino acid positions 24 and 116 and to complete loss of hydrolytic activity. The Turkish proband, in contrast, was homozygous for a G(1064)-->T substitution in exon 10 of the hLAL gene which converts the completely conserved glycine (GGG) residue at position 321 of the mature enzyme to tryptophan (TGG). In vitro expression of the hLAL(Gly(321)-->Trp) cDNA construct revealed that the amino acid replacement results in a more than 99% reduction of neutral lipid hydrolysis. The mutations provide new insights into the molecular basis of Wolman disease which is apparently more heterogeneous at the genetic level than cholesteryl ester storage disease.-Lohse, P., S. Maas, P. Lohse, A. C. Sewell, O. P. van Diggelen, and D. Seidel. Molecular defects underlying Wolman disease appear to be more heterogeneous than those resulting in cholesteryl ester storage disease.     

Regulation of progesterone biosynthesis in the human placenta by estradiol 17 beta and progesterone

The information available concerning the effects of chemotherapy administered during pregnancy is limited and consists of case reports and small series. A registry has been established at the National Cancer Institute, but there are currently only several hundred cases of neonates exposed to chemotherapy registered. All clinicians who care for women receiving chemotherapy during pregnancy should report those experiences to the National Cancer Institute to increase the data base. When chemotherapy is used during the embryogenesis period in the first trimester there is an increased rate of spontaneous abortion and major birth defects. The most toxic chemotherapeutic agents administered during pregnancy are methotrexate and aminopterin and should be avoided when possible, particularly during the first trimester. Pregnancy-related physiologic changes should be kept in mind when dosing and administering cytotoxic chemotherapy. The risk of fetal malformation when chemotherapy is administered during the second and third trimesters is probably not greater than background rate, but there may be a greater risk of stillbirth, fetal growth restriction, premature birth, and maternal and fetal myelosuppression. Breastfeeding should be avoided in women receiving chemotherapy.     

Immunohistochemical assessment of prostaglandin H-synthase in bovine endometrial biopsy samples collected throughout the oestrous cycle

In the Zhytomyr region, about 52,000 measurements of the 131I activity in thyroids were performed. On the basis of these measurements, individual doses have been assessed for the people monitored and age-dependent average doses have been estimated for those settlements with more than 11 direct measurements. In order to estimate the pattern of thyroid exposure in the Zhytomyr region, these doses have been interpolated or extrapolated to population groups who were not monitored during May-June 1986. For this purpose, a model has been developed based on a correlation between thyroid dose estimates with the 137Cs deposition and the co-ordinates of the settlements relative to Chernobyl. Collective doses of people who were born in the years 1968 to 1986 were calculated. The radiation-induced thyroid cancer incidence in the period 1991 to 1995 was assessed by subtracting the spontaneous incidence from the observed incidence. The result is considerably lower than that observed in longer periods after external exposures. Possible reasons for this difference are discussed.     

Immunohistochemical localization of ganciclovir in the human retina

Facial asymmetry (facedness) of selected academic faculty members was studied in relation to brain asymmetry and cognitive specialization. Comparisons of facedness were made among humanities faculty (H), faculty members of mathematics and physics (M-P), psychologists (P), and a group of randomly selected individuals (R). Facedness was defined in terms of the relative sizes (in square centimeters) of the two hemifaces. It was predicted that the four groups would show differences in facedness, namely, H, right face bias; M-P, left face bias; P, no bias; and R, no bias. The predictions were confirmed, and the results interpreted in terms of known differences in hemispheric specialization of cognitive functions as they relate to the dominant cognitive activity of each of the different groups. In view of the contralateral control of the two hemifaces (below the eyes) by the two hemispheres of the brain, the two sides of the face undergo differential muscular development, thus creating facial asymmetry. Other factors, such as gender, also may affect facial asymmetry. Suggestions for further research on facedness are discussed.     

Immunohistochemical localization of xanthine oxidase in human retina

Xanthine oxidase has been established as an important source of oxygen free radicals in ischemia-reperfusion injury. It has been localized in many different tissues such as heart and intestine, but it has not yet been localized in the eye. Xanthine oxidase was detected using immunohistochemistry on paraformaldehyde/glutaraldehyde fixed cryosections. Antibodies used included rabbit antibovine xanthine oxidase antibody and rabbit antihuman xanthine oxidase antibody. Xanthine oxidase was detected in the capillary endothelium cells of blood vessels in the retina of bovine and post mortem human eyes. Whole mount preparation of human retinas showed xanthine oxidase present throughout the small capillary network. Furthermore, whole mounts showed that xanthine oxidase was present in cones. This was confirmed by using mouse anticalbindin antibody for co-labelling. It is possible that xanthine oxidase can be a source of oxidative damage in the retina following ischemia-reperfusion injury.     

Prevention of endometrial hyperplasia by progesterone during long-term estradiol replacement: influence of bleeding pattern and secretory changes

OBJECTIVE: To determine the relative influences of induction of withdrawal bleedings secretory transformation, and reduction of mitosis in glands on prevention of endometrial hyperplasia during long-term hormonal replacement therapy. DESIGN: Observational expanded clinical case report. SETTING: Reproductive Endocrine Department of Hospital Necker, Paris, France, and Pathology Department of Women's Hospital, Los Angeles County and University of Southern California Medical Center, Los Angeles, California. PATIENTS: Postmenopausal women seeking treatment for symptomatic menopause. INTERVENTIONS: Endometrial biopsy and/or ambulatory hysteroscopy. MAIN OUTCOME MEASURE: Endometrial histology including progestational maturation patterns and glandular epithelial mitosis rates. Macroscopic endometrial appearance. RESULTS: The use of larger doses of E2 and P induced more marked secretory changes and more frequent withdrawal bleeding than the lower doses. There was no evidence of endometrial hyperplasia after 5 years of E2/P replacement therapy independently of bleeding pattern or progestational maturation. Consistent reduction of mitosis rates in glandular epithelium was found after 9 or more days of P administration in each cycle. CONCLUSIONS: Control of endometrial growth is mainly related to control of mitosis in glands by a relatively low doses of P. Induction of withdrawal bleeding and endometrial secretory transformation, which require larger doses of Progesterone, do not provide additional benefit for prevention of hyperplasia. Induction of amenorrhea with a relatively low dose of P may be offered to women seeking hormone replacement therapy with similar levels of safety.     

Immunohistochemical localization of nicotinic acetylcholine receptors in the guinea pig bowel and pancreas

Although nicotinic acetylcholine receptors (nAChRs) are known to be present on neural elements in both the bowel and the pancreas, the precise location of these receptors has not previously been determined. Immunocytochemistry, by using a rat monoclonal antibody (mAb35), which recognizes alpha-bungarotoxin (alpha-Bgt)-insensitive nAChRs, and a polyclonal antibody raised against the alpha-Bgt-sensitive receptor subunit, alpha7, was used to locate receptor protein in guinea pig gut and pancreas. mAb35-receptor (mAb35-R) immunoreactivity was abundant in both enteric plexuses, enterochromaffin cells, and pancreatic ganglia. Immunostaining was associated with the cell membrane, and clusters of mAb35-R were observed on cell somas and dendrites. Receptor immunoreactivity was also observed on terminals and axons, suggesting that a subset of nAChRs is presynaptic. Internal sites of mAb35-R were observed in permeabilized ganglia. Cells expressing the receptors were closely associated with ChAT-immunoreactive nerve fibers. In addition, the majority of ChAT-positive neurons expressed both cell surface and internal stores of mAb35-R. In the bowel, clusters of mAb35-R were present on the soma and dendrites of Dogiel type I motorneurons and secretomotor neurons. Receptors were detected at the plasma membrane of calbindin-immunoreactive myenteric neurons. In contrast, calbindin-immunoreactive submucosal neurons did not express cell surface mAb35-R, supporting the idea that they are sensory neurons. A subset of enteric neurons expressed both mAb35-R and glutamate receptor (GluR1) immunoreactivity. In the pancreas, mAb35-R immunoreactivity was only observed in ganglia. Alpha7-immunoreactivity was found on both enteric cell bodies and nerve fibers. Based on these results, it appears that visceral nAChRs are composed of at least four subunits and that both pre- and postsynaptic nAChRs are present in the gut and pancreas.