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PubMed1999

Intra- and intercellular Ca2+ signaling in retinal pigment epithelial cells during mechanical stimulation.

The intercellular communication (IC) was investigated between cultured rat retinal pigment epithelial (RPE) cells isolated from Long-Evans (LE) or dystrophic Royal College of Surgeons (RCS) rats and grown in solutions containing normal and high glucose concentrations, or after modulation of protein kinase C (PKC). This was performed by studying the conduction of the free Ca2+-concentration ([Ca2+]i) wave elicited by mechanical stimulation and by analyzing the fluorescence recovery after photobleaching (FRAP). Mechanical stimulation of LE-RPE cells triggers Ca2+ influx, mediated by stretch-sensitive cation channels followed by intracellular Ca2+ release. A regenerative [Ca2+]i wave was found with a lower propagation rate in RCS-RPE cells. This rate could be increased by PKC down-regulation. Mechanical stimulation caused a [Ca2+]i increase in the mechanically stimulated (MS) cell followed after a delay by a [Ca2+]i rise in the adjacent cell layers. The intercellular [Ca2+]i wave propagation could be blocked by gap junction blockers such as halothane or PKC activation. An inhibition of the [Ca2+]i-wave propagation similar to that induced by halothane could be observed in cells grown in solutions containing 14 mM or higher concentrations of glucose. PKC down-regulated cells grown in glucose-rich medium did not develop this inhibitory effect on gap junction communication (GJC). FRAP experiments confirmed that the observed changes were consistent with a PKC-mediated inhibitory effect of high glucose concentrations on GJC.

Animals↗
PubMed1998

[A case developing tetraplegia due to systemic lupus erythematosus which was remitted by a steroid].

The case reported here was a 58-year-old woman who was diagnosed as having systemic lupus erythematosus (SLE) in 1985 because she had erythema in the cheeks arthritis, a hematological abnormality (decreased white blood cell count), an immunological abnormality (LE-positive cells), and a positive result of antinuclear antibody test. Although the patient was once remitted by treatment with prednisolone (PSL) at 60 mg/day, and continuously received PSL at a maintenance dose of 2.5 mg/day, she was admitted in June 1996 by our hospital with chief complaints of fever and decreased muscular strength in the four extremities. At admission, she had symmetrical tetraplegia, which was peripherally predominant and severer in the lower extremities, and hypoesthesia accompanied by numbness. She was negative for anti-phospholipid antibody and showed no abnormality in cerebrospinal fluid examination. No lesions responsible for tetraplegia were detected at brain MRI, spinal MRI, or myelography. Because fever, multiple arthralgia, an increased erythrocyte sedimentation rate, a decreased lymphocyte count, hypocomplementemia, and a high immune complex level indicated the active stage of SLE (recurrence), she was given PSL at dose increased to 60 mg/day. After about 2 months, SLE was remitted and her tetraplegia and hypoesthesia was gradually improved thereafter. Although tetraplegia associated with peripheral nervous disorder, in which angitis occurring as a symptom of the active stage of SLE was thought to be Involved, was remitted by a steroid in out patient, no such cases have been reported in Japan, to our knowledge. Thus our patient was thought to be a very rare case of value.

Anti-Inflammatory Agents↗
PubMed1999

Estrogen receptor mRNA in uteri of normal estrous cycling and ovariectomized rats by in situ hybridization.

Biological effects of estrogen are mediated via its binding to the estrogen receptor (ER), the contents of its protein and mRNA varying during the estrous cycle. In the present study, the ERalpha mRNA expression in different cell components of the uterus was investigated in normal estrous cycling rats using nonisotopic in situ hybridization. Additionally, ovariectomized (OVX) rats treated with 17beta-estradiol (E2: 5 microg/kg, sc injection daily) were also investigated to clarify the effects of exogenous E2. At proestrus and diestrus, and especially the former, the luminal and glandular epithelial (LE and GE) cells were strongly positive, along with stromal cells beneath the luminal epithelium. At estrus, the expression was slightly diminished in LE cells, but almost completely lacking in GE cells. At metestrus, positive signals appeared again in GE cells. In the myometrium, ER mRNA was demonstrated to be constantly positive in all estrous cycle stages. OVX rat uteri underwent marked atrophy, but ER mRNA still remained in all cell types. After 2 consecutive days of E2 treatment, markedly increased intensity was observed, especially in LE and GE cells. The uteri of OVX rats treated with E2 for 14 days, however, showed slightly diminished expression, whereas the serum concentration of E2 was comparable to that in rats after 2 days. These results provide evidence that cell-type specific patterns of ER mRNA expression characterize the uteri of both normal estrous cycling rats and OVX rats after estrogen treatment.

Animals↗
PubMed1991

The effects of an atherogenic diet on macrophage/biomaterial interactions.

We previously reported that biomaterials differentially induced macrophages to secrete growth factors that mediate reendothelialization. The present study evaluates the effect of an atherogenic diet on macrophage/biomaterial interactions. Female New Zealand white rabbits were fed an atherogenic diet. Peritoneal macrophages were harvested from these as well as rabbits fed a normal diet and cultured in Minimum Essential Medium with platelet-poor serum. Dacron or polyglactin 910 were added to two of three conditions of both cell groups in passage 2. Conditioned media were collected weekly through week 15. Mitogenicity assays were performed with quiescent mouse embryonal (BALB/c3T3) fibroblasts, atherosclerotic rabbit aortic smooth muscle cells, and murine capillary lung (LE-II) endothelial cells. Mitogenic activity was assayed by scintillation counting of tritiated thymidine incorporation into deoxyribonucleic acid (DNA). Results showed increased mitogenic activity released by macrophages from atherosclerotic rabbits, in the absence of prosthetic material, when assayed against every cell line. In normal diet macrophages, polyglactin 910 stimulated mitogen release for every cell line, and Dacron yielded minimal mitogen release. In lipid diet macrophages polyglactin 910 slightly increased mitogen release for all three cell lines, whereas Dacron resulted in stimulation of DNA synthesis in smooth muscle cells and BALB/c3T3 cells but less DNA synthesis in LE-II cells than in control, no graft material, media. Western blotting demonstrated immunoreactivity to basic fibroblast growth factor in media from normal diet macrophages but only in the presence of polyglactin 910 or Dacron. Radioimmunoassay for platelet-derived growth factor B chain was negative in all groups, and polymerase chain reaction techniques to amplify transforming growth factor-beta messenger ribonucleic was negative. These data demonstrate the effect of in vivo dietary manipulation on macrophage activation as well as the effect of an atherogenic diet in modulating macrophage/biomaterial interactions. Additionally, different biomaterials differentially induce macrophages to release factors that stimulate and inhibit growth.

Animals↗
PubMed1991

Lewis antigen expression in benign and malignant tissues from RBC Le(a-b-) cancer patients.

Eight red blood cell (RBC) Le(a-b-) individuals were selected from a series of patients with bladder or colon cancer. Defined by the presence or absence of alpha 1-4-L-fucosyltransferase activity in saliva, four of these patients were characterized as non-genuine Lewis negative [RBC Le(a-b-) with alpha 1-4-L-fucosyltransferase activity in saliva], and four as genuine Lewis negative [RBC Le(a-b-) with no alpha 1-4-L-fucosyltransferase activity in saliva]. Stainings of paraffin embedded formalin fixed tissue sections for Lea and Leb antigens were performed by means of an indirect immunohistochemical method on all malignant and benign tissue previously removed from these eight patients. Leb antigens were always expressed independently of both the Lewis and the secretor status of the individual. Lea antigens, on the other hand, showed a different staining pattern. Although primarily expressed in non-genuine Le(a-b-) individuals, Lea antigens were expressed in genuine Le(a-b-) individuals as well--to a limited extent, but still detectable. Thus, these findings seem to show that the Lewis antigen expression is tissue dependent, and it is not possible to predict tissue Lewis antigen expression by merely examining erythrocytes or saliva.

ABO Blood-Group System↗
PubMed1988

The presence of CA19-9 in serum and saliva from Lewis blood-group negative cancer patients.

Eighteen cancer patients showed high levels of CA19-9 in sera, even though the blood-group phenotypes of their red blood cells were Le(a-b-). Seven of these patients (group I) were determined as Le(a-b-) from both red blood cells and saliva consistently, whereas eleven other patients (group II) secreted either Le(a) or Le(b) antigen in saliva and showed the expression of incompatible Lewis blood-group antigens. GDP-fucose: N-acetyl-glucosaminide alpha(1----4)-L-fucosyltransferase was demonstrated to be present in salivas from both group I and group II. These results suggest that a cancer-associated alteration of Lewis blood-group antigen expression occurs in cancer patients.

Antigens, Neoplasm↗
PubMed2006

Secreted phosphoprotein 1 (osteopontin) is expressed by stromal macrophages in cyclic and pregnant endometrium of mice, but is induced by estrogen in luminal epithelium during conceptus attachment for implantation.

Secreted phosphoprotein 1 (SPP1, osteopontin) is the most highly upregulated extracellular matrix/adhesion molecule/cytokine in the receptive phase human uterus, and Spp1 null mice manifest decreased pregnancy rates during mid-gestation as compared with wild-type counterparts. We hypothesize that Spp1 is required for proliferation, migration, survival, adhesion, and remodeling of cells at the conceptus-maternal interface. Our objective was to define the temporal/spatial distribution and steroid regulation of Spp1 in mouse uterus during estrous cycle and early gestation. In situ hybridization localized Spp1 to luminal epithelium (LE) and immune cells. LE expression was prominent at proestrus, decreased by estrus, and was nearly undetectable at diestrus. During pregnancy, Spp1 mRNA was not detected in LE until day 4.5 (day 1 = vaginal plug). Spp1-expressing immune cells were scattered within the endometrial stroma throughout the estrous cycle and early pregnancy. Immunoreactive Spp1 was prominent at the apical LE surface by day 4.5 of pregnancy and Spp1 protein was also co-localized with subsets of CD45-positive (leukocytes) and F4/80-positive (macrophages) cells. In ovariectomized mice, estrogen, but not progesterone, induced Spp1 mRNA, whereas estrogen plus progesterone did not induce Spp1 in LE. These results establish that estrogen regulates Spp1 in mouse LE and are the first to identify macrophages that produce Spp1 within the peri-implantation endometrium of any species. We suggest that Spp1 at the apical surface of LE provides a mechanism to bridge conceptus to LE during implantation, and that Spp1-positive macrophages within the stroma may be involved in uterine remodeling for conceptus invasion.

Animals↗
PubMed1992

The ELAM-1 ligand sialosyl-Le(X) is present on Langerhans cells isolated from stratified epithelium.

In this study we show the expression of the newly identified carbohydrate ligand, sialosyl-Le(X) on Langerhans cells. The receptor for sialosyl-Le(X) is the endothelial leukocyte adhesion molecule-1 (ELAM-1) present on activated endothelial cells. Using flow cytometry, Langerhans cells were selected due to positivity for an antibody against CD1a and low orthogonal light scatter. The CD1a antigen stained by the OKT6 antibody is considered a maturational marker of Langerhans cells in agreement with the specific labeling of dendritic cells in the epithelium only. Double immunostaining (OKT6/anti-sialosyl-Le(X)) using flow cytometry and immunohistochemistry demonstrated that almost all OKT6-positive cells in normal stratified epithelium expressed sialosyl-Le(X). Conversely, by immunohistochemistry of oral epithelium with acute inflammation, additional dendritic cells negative for OKT6 were found to express sialosyl-Le(X). In addition, sialosyl-Le(X)-positive but not OKT6-positive dendritic cells were found in the submucosa. These findings indicate that the carbohydrate antigen sialosyl-Le(X) is expressed earlier than the CD1a antigen in the maturation of the Langerhans cell lineage. Future studies should aim at investigating the importance of adhesion between sialosyl Le(X) and ELAM-1 in epithelial recruitment of Langerhans cells.

Antigens, CD↗
PubMed1991

Glycinergic control of [Leu5]enkephalin levels in chicken retina.

Retinal levels of [Leu5]enkephalin-like immunoreactivity (LE-LI) increase during the light and decrease during darkness, in vivo15. Intravitreal injection of the GABA antagonist picrotoxin had no effect on the accumulation of LE-LI during the light, suggesting the absence of significant GABAergic control over LE-LI cells. However, injection of the glycine antagonist strychnine, prevented the light-induced increase of retinal levels of LE-LI during 6 h exposure to light, indicating the presence of glycinergic control over the LE-LI neurons. When applied during the dark, strychnine increased the depletion of LE-LI by 34% compared to vehicle-injected eyes, suggesting that the LE-LI neurons receive some glycinergic input during the dark as well. The release of LE-LI from retinas superfused in vitro is depressed by exposing the preparation to light. Superfusing isolated retinas with physiological buffer containing picrotoxin (100 microM), GABA (50 mM), or the GABA agonists muscimol (100 microM), (+)-baclofen (200 microM), or 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP) (100 mM), had no effect on the efflux of LE-LI. Strychnine (100 mM) however increased the efflux of LE-LI by 64%, compared to the spontaneous efflux during the light. Glycine (15 and 50 mM) decreased the spontaneous efflux of LE-LI from retinas superfused in darkness by 44-48% and by 31% at 5 mM. These data are consistent with the results from pharmacological manipulations in vivo. We conclude that the LE-LI amacrine cells are under inhibitory control from glycinergic but not from GABAergic neurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

[Sialosyl-Le(a) and Sialosyl-Le(x) antigens in adhesion of cancer cells and tumor progression].

Sialosyl-Le(a) and sialosyl-Le(x) are tumor-associated carbohydrate antigens present in different types of human tumors. They are commonly found on the cell surface of a variety of adenocarcinomas such as lung cancer, gastric cancer, pancreatic cancer and colorectal cancer, and in serum of cancer patients. Both antigens have been proposed as important diagnostic markers and they are used in detecting and monitoring of these diseases. Recently, it has been shown that sialosyl-Le(a) and sialosyl-Le(x) carbohydrate structures are ligands for selectins, newly described family of adhesion molecules. Selectins function as lymphocyte-homing and leukocyte enrollment receptors, or as activation dependent cell surface receptors of platelets and endothelial cells. Several lines of evidence suggest that sialosyl-Le(a) and sialosyl-Le(x) are responsible for adhesion of human cancer cells to endothelium. It has been shown that E-selectin and P-selectin present on endothelial cells mediate these interactions. The mentioned facts suggest that selectins and their carbohydrate ligands can play an important role in a selective homing of tumor cells during metastasis.

Animals↗
PubMed2003

[Characterization of anti-DNA antibody on IgG3-kappa type M-protein in a patient with rheumatoid arthritis].

A 66 year-old male patient with rheumatoid arthritis, was admitted due to bronchial pneumonia. Anti-nuclear antibody and anti-DNA antibody were both detected in the patient's serum and many LE-like cells were found in the pleural effusion. In addition, M-proteins(IgG-kappa type, IgM-kappa type) were detected in both serum and pleural effusion by immunofixation electrophoresis. Purified IgG fraction by gel filtration(Superdex 200HR) showed high titer of anti-nuclear antibody(x160) and anti-DNA(39.2 AU/ml) antibody. Furthermore, anti-DNA antibody was detected in the fraction III(fraction of M-protein) by ion-exchange chromatography. In conclusion, these results indicate that M-protein(IgG3-kappa type) contains anti-DNA antibody and it is suggests that M-protein induces the production of LE like cell in this case.

Aged↗
PubMed1985

The nature of the defect in experimental allergic encephalomyelitis (EAE)-resistant Lewis (Le-R) rats.

Recently, a colony of Lewis rats has been described which is resistant to experimental allergic encephalomyelitis (EAE). These rats, termed Le-R, are still histocompatible with other Lewis rats. The genetic defect which results in EAE-resistance was shown not to be linked to the RT1.B (Ir) region of the MHC. Myelin basic protein (BP)-sensitization of Le-R rats induces cells capable of mounting a proliferative response to BP in culture but incapable of transferring EAE after culture with BP. The present study demonstrates that the latter deficiency can be overcome either by incorporating lipopolysaccharide (LPS) in the BP-culture medium or by simultaneous transfer of LPS-activated antigen-nonspecific spleen cells with the BP-sensitized cells. The BP-sensitized Le-R cells fail to transfer EAE due to their inability to initiate lesions in the CNS. LPS, working through an antigen-nonspecific cell or cell products, can correct the defect in the Le-R cells such that the antigen-specific cells become capable of initiating CNS lesions which lead to development of clinical EAE.

Animals↗
PubMed1991

Characterization of mucin-type oligosaccharides with the sialyl-Le(a) structure from human colorectal adenocarcinoma cells.

Oligosaccharides with the sialyl-Le(a) structure have been isolated on an affinity column of a monoclonal antibody, MSW 113, from mucin-type glycoproteins derived from the surfaces of SW 1116 and LS 180 cells, and their secretions. The oligosaccharides were polydisperse with respect to molecular size, the oligosaccharides derived from glycoproteins in culture media being larger than those in cell lysates, as assessed by gel filtration. Some of the oligosaccharides were susceptible to degradation by endo-beta-galactosidase (E. freundii), as judged from the change in the gel filtration pattern. These results indicate that oligosaccharides with the sialyl-Le(a) structure derived from mucin-type glycoproteins produced by human colonic cancer cells are extremely large in size and complex in structure, and that some of them contain the poly-N-acetyllactosamine structure.

Adenocarcinoma↗
PubMed1994

Specificity analysis of blood group Lewis-y (Le(y)) antibodies generatedagainst synthetic and natural Le(y) determinants.

Le(y)-reactive monoclonal antibodies (mAbs) were generated in mice by immunization with synthetic Le(y) neoglycoproteins or with Le(y)-expressing cells. Serological analysis indicated that mAbs raised against synthetic Le(y) (i) reacted strongly with synthetic Le(y) but poorly with natural Le(y), (ii) cross-reacted with Le(x) or H-type 2 structures, and (iii) were IgG1, IgG2a, or IgG2b. mAbs raised against Le(y)-expressing cells (i) reacted with both synthetic Le(y) and natural Le(y), (ii) were of two types: cross-reactive with Le(x) or H-type 2 structures or specific for Le(y), and (iii) were IgM or IgG3. One of the mAbs raised against natural Le(y), mAb 3S193 (IgG3), showed high specificity for Le(y) in ELISA tests with synthetic Le(y) and Le(y) containing glycoproteins and glycolipids; it also reacted strongly in rosetting assays and cytotoxic tests with Le(y)-expressing cells. mAb 3S193 did not lyse O, A, AB, and B human erythrocytes in the presence of human complement. In flow cytometry, there was weak reactivity with granulocytes, a reactivity also observed with two previously described highly specific Le(y) mouse mAbs--BR55-2 (IgG3) and B3 (IgG1). A humanized version of mAb 3S193 has been constructed, and the specificity pattern and reactivity for Le(y) remain very similar to mouse mAb 3S193.

Animals↗
PubMed1995

The pattern of mutations induced by neocarzinostatin and methyl methanesulfonate in the ataxia telangiectasia-like Chinese hamster cell line V-E5.

The Chinese hamster cell line V-E5 is a mutant cell line isolated from V79 cells. The phenotypic characteristics of V-E5 strongly resemble those of cells from patients suffering from the genomic instability syndrome ataxia telangiectasia. In order to further characterize the mutant cell line and to get insight into the underlying genetic defect we compared the clastogenic and mutagenic effects of neocarzinostatin (NCS) and methyl methanesulfonate (MMS) in V-E5 and V79 wild-type cells (V79-LE). V-E5 cells were 2-3 times more sensitive to the cytotoxic effect of NCS or MMS. The clastogenic action of NCS was characterized by the predominant induction of chromosome breaks and dicentrics in both cell lines, whereas MMS mainly induced chromatid-type aberrations. The frequency of mutations induced by NCS as well as MMS was slightly enhanced in V-E5 cells compared to V79 cells treated with the same dose. However, the mutant cell line was found to be hypomutable when considering the same survival level as in the parental cell line. Molecular analysis of mutants induced by NCS revealed a high frequency of total deletions of the hprt gene in both cell lines. In contrast, among MMS-induced mutations only 11% deletion mutations were found in V79-LE, whereas in V-E5 MMS-induced deletions were seen in 52% of the hprt-deficient mutants. These results are discussed with respect to a possible relation between genomic instability, cell cycle control and mutational spectra.

Animals↗
PubMed1980

The dorsal nucleus of the lateral lemniscus in the cat: neuronal types and their distributions.

The normal population of neurons and their distributions within the dorsal nucleus of the lateral lemniscus were studied in both Nissl-stained celloidin and frozen sections and in Golgi impregnations from brains of mature cats. According to axial measurements of somata in Nissl-stained material, neurons of the dorsal nucleus of the lateral lemniscus (DNLL) were classified by width:length ratio (r) into round (0.80 less than or equal to r less than or equal to 1.0), ovoid (0.65 < r < 0.80), or elongate (r less than or equal to 0.65) types. These same neurons could also be classed by average diameter (Dm) as large (Dm greater than or equal to 22), medium (12.0 less than or equal to Dm < 22.0), or small (Dm < 12.0). A combination of data on ratios (shape) and average diameters (size) provided the following possible categories of Nissl-stained, DNLL neurons: large round (LR), large ovoid (LO), large elongate (LE), medium round (MR), medium ovoid (MO), medium elongate (ME), small round (SR), small ovoid (SQ), and small elongate (SE). Very few small cells were found, however. Quantitative studies of the distributions of cell type within the whole DNLL showed (1) most medium-sized and most LE cells in the caudal third of the DNLL and (2) most LO and LR cells dorsally located in the rostral third of the DNLL. There were progressively more large and more round types along the caudal-to-rostral axis. In Golgi impregnations of the DNLL, all medium and large cell types, but no small cell types (defined in the Nissl study) were found. Golgi material showed (1) subdivisions of the LO class into vertical (LOV) and horizontal (LOH) types, and (2) radiate (MRR) and oriented (MRO) subclasses of MR neurons according to dendritic arbor and cytology, orientation within the DNLL, and axonal morphology. Examples from all classes of large cells (particularly, LE cells) could have ventrally directed axons. These ventrally directed axons might be efferents to the cochlear nucleus, known from our previous work. A strong horizontal orientation of most DNLL cell somata and dendrites, shown in both our Nissl and Golgi material, is discussed in relation to known inputs to the DNLL. Correlations of our morphological findings with limited electrophysiological data on the DNLL are also discussed.

Animals↗