GnRH agonist-suppressed expression of nitric oxide synthases and generation of peroxynitrite in adenomyosis

Because overproduction of nitric oxide (NO) and peroxynitrite is known to cause tissue injury, the expression of NO synthases (NOS) and generation of peroxynitrite were investigated in adenomyosis. Immunoreactivities to endothelial and inducible NOS demonstrated phase-dependent changes in normal endometrium, and in eutopic endometrium of adenomyosis. However, NOS were expressed throughout the menstrual cycle in ectopic endometrium from the majority of patients with adenomyosis. Nitrotyrosine, a footprint of peroxynitrite, was detected concomitantly with NOS protein. This suggested that high doses of NO and superoxide are produced in the ectopic endometrium, presumably by stimulation with bioactive molecules such as cytokines and growth factors. The expression of NOS and generation of peroxynitrite were markedly reduced by administration of gonadotrophin-releasing hormone agonists (GnRHa). The suppression of serum concentrations of nitrite/nitrate, stable metabolites of NO, by long-term administration of GnRHa was also demonstrated. The suppression of synthesis of NO and/or peroxynitrite may be part of both the therapeutic and adverse effects of GnRHa therapy.     

Transgenic tobacco cells producing the human monoclonal antibody to hepatitis B virus surface antigen

The recombinant human monoclonal antibody (MAb) against hepatitis B virus (HBV) surface antigen (HBsAg) was expressed in tobacco suspension cultures. The parental CL4MAb was produced by the Epstein-Barr virus (EBV) transformed human cell line TAPC301-CL4. The CL4MAb cDNA was introduced into tobacco suspension cells by Agrobacterium mediated transformation. The monoclonal antibodies (MAbs), B294 and B303, which were derived from CL4 and subsequently produced in plant cells were selected for study. After purification on Protein A columns, B294 and B303 MAbs had anti-HBs relative affinity constants similar to the parental CL4MAb. B303 MAb interacted with cell surface HBsAgs and showed complement-dependent cytotoxicity in a manner that was similar to anti-HBs human immunoglobulins (HBIg) that are used clinically. The results of this study point to the feasibility of producing MAbs to HBsAg in plants as an alternative to HBIg.     

Expression of Lewisx sugar structure in the liver metastasis of mouse colon carcinoma (colon 26) cells

Immunohistochemical aspects of the process of experimentally induced metastasis were examined by light and electron microscopy employing a series of labeled carbohydrate-specific monoclonal antibodies as probes. Liver metastasis was induced by injecting mouse colon carcinoma cell (colon 26) into the spleen of Balb/c mice. Labeled anti-Lewis^x (Le^x) antibody stained the metastasized colon 26 cells strongly compared with the heterogeneous and faint staining in non-metastasized tumor foci in the spleen or in the subcutaneous space. Other antibodies having specificities for Lewis-related antigens other than Le^x, e.g. those against Le^y, Le^a, Le^b, sialyl Le^x and sialyl Le^a, did not show any differences in binding between metastasized cells and non-metastasized tumor foci. Immunoelectron microscopy revealed the expression of Le^x-carbohydrate in the plasma membranes as well as in the intercellular spaces of metastasized colon 26 cells in the liver. Based on these results, it is likely that sugar chains containing the Le^x-carbohydrate structure are involved in the interactions between colon 26 cells and hepatic cells during the process of liver metastasis.     

Effects of blinding,olfactory bulbectomy,and pinealectomy on prolactin and growth hormone cells of the rat,with special reference to ultrastructure

The effects of combined blinding and olfactory bulbectomy alone or with pinealectomy on body growth, and pituitary and serum levels of growth hormone and prolactin were examined in young female rats. The animals were 23–24 days old at the time of the operations, and they were sacrificed 52 days later. Blind, anosmic rats had elevated serum and depressed pituitary stores of prolactin. The blind, anosmic rats which were also pinealectomized showed prolactin levels which were intermediate between those of the blind, anosmic and sham control animals. On the other hand, combined blindness and anosmia decreased serum growth hormone levels and again pinealectomy prevented this effect. The observed changes in the ultrastructure of prolactin and growth hormone cells were well correlated with these alterations in hormone levels.     

Effects of estrogen replacement on insulin-like growth factor I concentrations in serum and bone tissue and on interleukin 1 secretion from spleen macrophages in oophorectomized rats

Summary Oophorectomy (OOX) has been known to increase bone turnover, but its precise mechanism is not fully understood. In order to further investigate the mechanism, we determined insulinlike growth factor I (IGF-I) concentrations in serum and bone tissue and interleukin 1 (IL-1) release from spleen macrophages in oophorectomized rats because it has been demonstrated that IGF-I stimulates bone formation and IL-1 stimulates bone resorption.Female 8-week-old Wistar rats were divided into four groups: (1) control, (2) OOX, (3) OOX given estradiol, and (4) control given estradiol. Ten g/kg of 17-estradiol was given daily by subcutaneous injection. After 5 weeks of treatment, IGF-I concentrations in the extract from right femur and in serum were determined by specific radioimmunoassay. IL-1 activity released from lipopolysaccharide (LPS)-stimulated spleen macrophages was determined by bioassay. IGF-I contents in the femur and IGF-I concentrations in serum in oophorectomized rats were significantly higher than those in control rats. Treatment by estradiol inhibited the increase in IGF-I concentrations both in femur and in serum. IL-1 release from LPS-stimulated spleen macrophages in oophorectomized rats was increased, and treatment by estradiol also inhibited the stimulated IL-1 release. The ash weights and the calcium contents of left femur in oophorectomized rats were lower than those in control rats. These results suggest that both IGF-I and IL-1 may be involved in the mechanism of the regulation of bone turnover in oophorectomized rats.Abbreviations IGF insulinlike growth factor - IL interleukin - OOX oophorectomy - LPS lipopolysaccharide - PTH parathyroid hormone - BGP bone gla protein - MEM minimum essential medium - FCS fetal calf serum     

Interleukin 2 receptor gamma chain (gamma(c)) knockout mice show less regularity in estrous cycle but achieve normal pregnancy without fetal compromise

PROBLEM: Implication of cytokines in pregnancy suggested but remains to be established. We studied the effect of cytokines on reproductive functions such as ovulation and pregnancy, with mutant mice lacking interleukin 2 receptor gamma (IL-2Rgamma), the so-called common gamma chain (gamma(c)), which is shared among receptors for multiple cytokines such as IL-2, IL-4, IL-7, IL-9 and IL-15. METHOD OF STUDY: Regularities of estrous cycles were observed by vaginal smear. Ovaries stimulated with postmenopausal serum gonadotropin (PMSG) were examined for the ovarian capacities. The uteri at 13 days of gestation were used for histological analysis of the maternal-fetal interface. RESULTS: The estrous cycles in gamma(c) knockout (gamma(c) KO) mice were irregular compared with wild-type mice, although, the mutants could become pregnant. No uterine natural killer (uNK) cell was found in the uterus at 13 days of pregnancy, and poor decidual formation and thickness of blood vessel walls were observed. Apparent differences were not seen in the numbers and weight of the fetuses between wild type and mutant animals, and fetuses were not compromised throughout pregnancy. CONCLUSIONS: The gamma(c) KO mice showed irregular estrous cycle but they could carry out normal pregnancy despite lacking uNK and cytokines actions of IL-2, 4, 7, 9 and 15.     

Diffuse pulmonary hamartoma: a case report

Pulmonary hamartoma usually occurs as a benign, well-circumscribed single nodule in the lung parenchyma. We report on a unique case of hamartoma that extended along the bronchial tree, formed endobronchial polypoid lesions, and expanded into the lung parenchyma. The patient, a 48-year-old man, was admitted with dyspnea and chest pain. A transbronchial biopsy was performed on a tumorous lesion that was diagnosed histologically as a hamartoma. As this lesion was found to be growing diffusely along the bronchial tree, a left pneumonectomy was performed. Gross examination showed that yellowish soft tissue had surrounded the bronchial tree and extended into the lung parenchyma. Histologically, the lesion contained mainly mature adipose tissue, cartilage, and muscle tissue with a minor component of short spindle cells in a myxomatous matrix. The patient was diagnosed as having diffuse pulmonary hamartoma.     

Dispensability of Jak1 tyrosine kinase for interleukin-2-induced cell growth signaling in a human T cell line

The tyrosine kinases Jak1 and Jak3 are known to be associated with the β and γ chains of interleukin-2 receptor (IL-2R). They are activated by stimulation with IL-2, IL-4, IL-7, IL-9, or IL-15, receptors of which share the γ chain of the IL-2R. We have obtained direct evidence of Jak1 association with the α chains of receptors for IL-4, IL-7 and IL-9 and with the β chain of IL-2R, which is also common to the IL-15R. Furthermore, we have prepared mutant IL-2R β chains with a mutation in the box 1 region, which is conserved among the IL-2R β chain and the α chains of the other cytokine receptors sharing the IL-2R γ chain. Using MOLT-4 transfectants with the mutant β chains, we found that two conserved proline residues within the box 1 region are essentially involved in association with Jak1. The MOLT-4 transfectants with the mutant β chains lacking Jak1 association showed IL-2 responsiveness, in terms of activation of Jak3 and Stat5 and induction of cell growth, indicating that Jak1 is dispensable for IL-2-mediated cell growth signaling and that Jak1 activation is not required for activation of Jak3 and Stat5 in the MOLT-4 transfectants.     

Evolution of ribosomal DNA unit on the X chromosome independent of autosomal units in the liverwort Marchantia polymorpha

In the haploid dioecious liverwort, Marchantia polymorpha, the X chromosome, but not the Y, carries a cluster of ribosomal RNA genes (rDNAs). Here we show that sequences of 5S, 17S, 5.8S and 26S rDNAs are highly conserved (>99% identity) between the X chromosomal and autosomal rDNA repeat units, but the intergenic spacer sequences differ considerably. The most prominent difference is the presence of a 615-bp DNA fragment in the intergenic spacer, X615, which has accumulated predominantly in the rDNA cluster of the X chromosome. These observations suggest that the rDNA repeat unit on the X chromosome evolved independently of that on autosomes, incorporating sex chromosome-specific sequences. This revised version was published online in July 2006 with corrections to the Cover Date.