Biomedical subjects

M Sakaguchi

Publications and source records attributed to M Sakaguchi.

At least 163 records · Page 9Linked to original sources

PubMed1999

Development of a sensitive, specific reverse transcriptase polymerase chain reaction-based assay for epithelial tumour cells in effusions.

We developed a sensitive and specific method for the detection of epithelial cancer cells in effusions with a two-stage molecular-based assay which combined enrichment for cancer cells by immunomagnetic bead selection and reverse transcriptase polymerase chain reaction (RT-PCR) detection of epithelial glycoprotein 2 (EGP-2) RNA. Preliminary experiments indicated that immunobead selection was essential to avoid occasional false-positive RT-PCR results, and this method detected ten breast cancer cells electively added to 10(7) cytologically negative effusion cells. We studied 110 cases of pleural (n = 68) and peritoneal (n = 42) effusions (30 from patients with known carcinoma and 80 from those without known carcinoma), and the results were compared with cytological findings. Of 18 effusions that were cytologically positive or suspicious for malignant cells, 17 (94%) were positive for EGP-2 RNA (the one negative sample was from a patient who recently received combination chemotherapy). Of 92 cytologically negative samples, 11 (12%) were positive for EGP-2, including six patients with a history of previous or current carcinoma. Our method appears to be highly specific and increases the sensitivity of detection of malignant cells; it may be a useful adjunct to routine cytopathological examination.

Ascitic Fluid↗

PubMed1999

IgE reactivity and cross-reactivity of Japanese monkeys (Macaca fuscata) to Japanese cedar (Cryptomeria japonica) and cypress (Chamaecyparis obtusa) pollen allergens.

BACKGROUND: The natural occurrence of Japanese cedar (Cryptomeria japonica, CJ) pollinosis has been reported in Japanese monkeys (Macaca fuscata). However, the reactivity to Japanese cypress (Chamaecyparis obtusa, CO) pollen allergens in these monkeys has not yet been reported. OBJECTIVES: The present study was designed to investigate the reactivity to CO pollen allergens in monkeys sensitized to CJ pollen allergens. METHODS: Serum samples from 40 monkeys naturally sensitized to CJ pollen allergens were collected from four troops. We measured the specific IgE to CO pollen allergens and examined the reactivity to the allergens by intradermal test. Cross-reactivity between CJ and CO pollen allergens was examined by ELISA inhibition method. Furthermore, we examined the sensitivity to the allergens by histamine release assay from leucocytes. RESULTS: All 40 monkeys had specific IgE to crude and purified major allergens (Cha o 1) of CO pollen. The monkeys showed a positive reaction to CO pollen allergens in the intradermal test. Allergenic cross-reactivity between Cha o 1 and Cry j 1 (a major allergen in CJ pollen) was also observed. Specific histamine release to both the major allergens was noted in two monkeys with CJ pollinosis. CONCLUSION: Japanese monkeys sensitized to Japanese cedar pollen allergens also demonstrate reactivity to Japanese cypress pollen allergens.

Allergens↗

PubMed1999

Sensitization to grass, ragweed, mugwort pollen allergens in Japanese monkeys (Macaca fuscata): preliminary report.

BACKGROUND: The natural occurrence of Japanese cedar (CJ, Cryptomeria japonica) pollinosis has been reported in Japanese monkeys (Macaca fuscata). Furthermore, most of these monkeys with CJ pollinosis have immunoglobulin (Ig) E sensitization to Japanese cypress (Chamaecyparis obtusa) pollen. However, specific IgE to other pollens has not yet been reported. OBJECTIVES: The present study was designed to investigate IgE sensitization of Japanese monkeys to grass, ragweed, and mugwort pollen. METHODS: Serum samples from 47 monkeys as a general population in one troop were collected at random. We measured specific IgE to grass, ragweed and mugwort pollen. Next, 10 monkeys with CJ pollinosis from the same troop were also examined for their IgE sensitization to grass, ragweed, and mugwort pollen. RESULTS: Of 47 monkeys, 13 (28%) had specific IgE to CJ pollen, 15 (32%) to grass pollen, five (11%) to ragweed pollen, and three (6%) to mugwort pollen. Furthermore, CJ pollinosis monkeys seemed to be sensitized to these pollen allergens with higher frequency; of 10 monkeys, 10 (100%) had specific IgE to CJ pollen, six (60%) to grass pollen, four (40%) to ragweed pollen, and two (20%) to mugwort pollen. CONCLUSION: Japanese monkeys had specific IgE to grass, ragweed, and mugwort pollen in addition to CJ pollen.

Allergens↗

PubMed1999

Reactivity of the immunoglobulin E in bovine gelatin-sensitive children to gelatins from various animals.

It has been reported that most children who showed anaphylaxis to measles, mumps and rubella vaccines containing bovine gelatin as a stabilizer have anti-bovine gelatin IgE. The present study was designed to investigate the reactivity of IgE in bovine gelatin-sensitive children to gelatins from various animals, and the antigenic cross-reactivity between the gelatins. Serum samples taken from 10 children who showed anaphylaxis to vaccines containing bovine gelatin were used in this study. The level of anti-bovine gelatin IgE in these serum samples ranged from 11.0 to 251 Ua/ml. The IgE in most of the children reacted to kangaroo and mouse gelatins, to which they had had little or no exposure as a food or a vaccine stabilizer. The IgE binding to kangaroo and mouse gelatins was completely inhibited by bovine gelatin, whereas reciprocal inhibition was not complete, indicating that antigenic cross-reactivity is present between the mammalian gelatins. Only one child had strong IgE reactivity to fish gelatins, and this reactivity was not inhibited by bovine gelatin, indicating that no antigenic cross-reactivity exists between bovine and fish gelatins. Most of the children who displayed sensitivity to bovine gelatin showed IgE reactivity to other mammalian gelatins. This reactivity may be due primarily to the antigenic cross-reactivity between mammalian gelatins.

Anaphylaxis↗

PubMed1999

The incidence of Japanese cedar pollinosis and sensitization to the pollen allergens among Japanese monkeys in a troop.

The natural occurrence of Japanese cedar (Cryptomeria japonica; CJ) pollinosis has been reported in Japanese monkeys (Macaca fuscata), an appropriate animal model for developing antipollinosis therapies. However, there has been no study on the incidence of Japanese cedar pollinosis in monkeys. To evaluate the incidence of CJ pollinosis in Japanese monkeys, we investigated the presence of pollinosis symptoms among monkeys in a troop, and the response to CJ allergens in pollinosis monkeys. We examined the presence of pollinosis symptoms in 272 monkeys in a troop throughout the CJ pollination season (February to April). Of the 272 monkeys, 21 (7.7%) showed pollinosis symptoms during the CJ pollen season. Blood samples were taken from the 21 monkeys that showed pollinosis symptoms and were tested for the presence of immunoglobulin E (IgE) antibody for CJ allergens. All 21 monkeys with CJ pollinosis had anti-CJ IgE. Of the 21 monkeys, peripheral blood mononuclear cells (PBMC) could be taken from 12, all of which showed CJ allergen-specific PBMC proliferation. The incidence of CJ pollinosis in a troop was 7.7%. The monkeys with CJ pollinosis demonstrated specific IgE and PBMC proliferation for CJ allergens.

Allergens↗

PubMed1999

The incidence of japanese cedar pollinosis and sensitization to the pollen allergens among Japanese monkeys in a troop.

Allergens↗

PubMed1999

Effects of a new 12-lipoxygenase inhibitor, daphnodorin A, on angiotensin II-induced vascular contraction in hamster.

We studied whether lipoxygenase inhibition suppressed angiotensin II-induced vascular contraction. In the present study, we used a new 12-lipoxygenase inhibitor, daphnodorin A, and an analogue of daphnodorin A, daphnodorin B, which has no inhibitory effects on 12-lipoxygenase. Daphnodorin A at 30 microM and 100 microM significantly suppressed the contractile responses induced by angiotensin 11 (3 x 10(-8) M) in isolated hamster aorta, while daphnodorin B up to 100 microM did not affect the responses. These results suggest that daphnodorin A, but not daphnodorin B, may suppress angiotensin II-induced vascular contractile responses through the inhibition of 12-lipoxygenase.

Angiotensin II↗

PubMed1999

Novel gelatinolytic activities in rat organs.

Novel gelatinolytic activities in both latent and active forms were detected in the normal organs of rat by gelatin zymography. Multiple active bands were detected in the extracts from the skin, jejunum, muscle, and kidney without any activation. These activities were inhibited by 1,10-phenanthroline or leupeptin, nor by E64, suggesting that these activities were derived from metallo-proteinases or serine-proteinases. Some gelatinolytic active bands were newly induced or enhanced by p-aminophenylmercuric acetate. These results suggest that matrix degrading activities due to metallo- and serine-proteinases were constitutively expressed in various rat normal organs.

Animals↗

PubMed1999

Measurements of biological contaminants and particulate matter inside a dwelling in Japan.

To investigate the biological contaminant levels inside dwellings in Japan, the authors measured fungal indices, airborne mite allergens (Der 1 and Der 2), and suspended particulate matter (SPM). Measurements were continuously conducted at one dwelling located in Yokosuka-city, Kanagawa. The biosensors for fungal index were set at 12 points inside and 2 points outside the dwelling. An SPM sampler was placed in the living room, and samplers for collecting allergens were placed in the living room and in the washroom. The peak level of the fungal index was recorded during July, which is the rainy season in Japan. The highest value in the dwelling was 6.9 in the toilet, and the second highest was 6.6 near a window in the living room that faces southeast, and behind the sofa. The highest monthly SPM level was 21.5 micrograms/m3 during October, and the highest monthly allergen level (Der 1) in the living room was 25.7 pg/m3 during November. A positive relationship between the fungal index and mite allergens was not observed, and the levels of contaminants were not as those found in other studies. However, further study to investigate the distribution of biological contaminants is needed because indoor temperature and relative humidity are comparatively high.

Air Pollution, Indoor↗

PubMed1999

Transferrin synthesized in cultured human fibroblasts is associated with tubulins and has iron binding capacity.

In a previous report (13), using immunocytochemical and fluorescence-labeling techniques, we demonstrated that transferrin is synthesized in cultured human fibroblasts and that it is associated with tubulins in the cells. These morphological findings led us to attempt to elaborate those issues in more detail by biochemical methods. In this report, we were able to prove the association of transferrin produced in cells with tubulins. In addition, the transferrin associated with tubulins was found to bind to iron. These results suggest that endogenous transferrin plays a role in preventing damage caused by free radicals which can be induced by the interaction of iron with the hydrogen peroxide produced in cells.

Cells, Cultured↗

PubMed1999

Effects of a lipoxygenase inhibitor, panaxynol, on vascular contraction induced by angiotensin II.

We investigated whether a lipoxygenase inhibitor, panaxynol, affected the vascular contraction induced by angiotensin (Ang) II and the mean arterial pressure in spontaneously hypertensive rats (SHR). Panaxynol suppressed dose-dependently the contractile responses induced by 30 nM Ang II in isolated intact and endothelial cell-denuded aorta in the hamster. IC50 values in the intact and endothelial cell-denuded aorta were 23 and 20 microM, respectively. In SHR, the mean arterial pressure after injection of 30 and 60 mg/kg panaxynol was reduced, and the maximum hypotensive values were 23 and 48 mmHg, respectively. Thus, lipoxygenase products may affect the renin-angiotensin system.

Alkynes↗

PubMed1999

Effects of Sho-seiryu-to on experimental allergic rhinitis in guinea pigs.

The effects of Sho-seiryu-to, an antiallergic Kampo medicine, on experimental allergic rhinitis were investigated in actively sensitized guinea pigs. The number of sneezes and scratches by the animals after a topical antigen challenge was significantly inhibited by pretreatment with Sho-seiryu-to (1000 mg/kg per os p.o.). The antigen-induced eosinophil infiltration in the nasal mucosa was significantly inhibited by Sho-seiryu-to (1000 mg/kg p.o.). Sho-seiryu-to (100 mg/kg p.o.) also reduced the increase in dye leakage to the nasal cavity induced by the antigen challenge and the antigen-induced decrease in volume of the nasal cavity was inhibited. Moreover, Sho-seiryu-to (1000 mg/kg p.o.) suppressed the volume change in the nasal cavity induced by leukotriene D4. These results demonstrate that Sho-seiryu-to inhibits experimental allergic rhinitis in guinea pigs, confirming that the agent may be beneficial for the treatment of allergic rhinitis.

Animals↗

PubMed1999

[Leiomyoma of the lung: a case report].

An asymptomatic 43-year-old female was admitted to the Hospital because of an abnormal nodule in the left lung field on screening chest X-ray. Chest CT showed a tumor shadow mass (4 x 3 cm) in the left lower lobe. Bronchofiberscopy revealed an endobronchial polypoid mass obstructing the lower lobe bronchus at the orifice of left B10. Lt lower lobectomy was performed. Histopathological examination of the specimen showed the tumor (4.5 x 3.6 x 3.6 cm) developing not only within the tracheal lumen, but also in the lung tissue. Because immunohistochemical staining for desmin and alpha-smooth muscle actin was positive, we diagnosed leiomyoma. As MRI revealed multiple uterine masses after operation, we discussed the diagnostic problems in the relation to metastasizing leiomyoma of the uterus.

Adult↗

PubMed1998

Characterization of paired tumor and non-tumor cell lines established from patients with breast cancer.

The goal of our study was to develop a panel of tumor cell lines along with paired non-malignant cell lines or strains collected from breast cancers, predominantly primary tumors. From a total of 189 breast tumor samples consisting of 177 primary tumors and 12 metastatic tissues, we established 21 human breast tumor cell lines that included 18 cell lines derived from primary tumors and 3 derived from metastatic lesions. Cell lines included those from patients with germline BRCA1 and FHIT gene mutations and others with possible genetic predisposition. For 19 tumor cell lines, we also established one or more corresponding non-malignant cell strains or B lymphoblastoid (BL) lines, which included 16 BL lines and 7 breast epithelial (2) or stromal (5) cell strains. The present report describes clinical, pathological and molecular information regarding the normal and tumor tissue sources along with relevant personal information and familial medical history. Analysis of the breast tumor cell lines indicated that most of the cell lines had the following features: they were derived from large tumors with or without axillary node metastases; were aneuploid and exhibited a moderate to poorly differentiated phenotype; were estrogen receptor (ER)- and progesterone receptor (PR)-negative; and overexpressed p53 and HER2/neu proteins. Of 13 patients with primary breast cancers receiving curative intent mastectomies, 7 were dead after a mean period of 10 months. Our panel of paired tumor and non-malignant cell lines should provide important new reagents for breast cancer research.

Adult↗

PubMed1998

Assessment of topogenic functions of anticipated transmembrane segments of human band 3.

Band 3 protein is a typical multispanning membrane protein whose membrane topology has been extensively studied from various protein chemical approaches. To clarify the membrane topogenesis of this multispanning protein on the endoplasmic reticulum, the topogenic functions of the anticipated transmembrane segments were individually assessed in an in vitro system using two series of model proteins in which each segment was placed in either a "stop-transfer" context or a "translocation initiation" context. They were expressed in a cell-free system containing rough microsomal membranes, and their topologies were evaluated by taking advantage of either sensitivity to protease or accessibility to N-glycosylation. We found that some segments seem to possess insufficient topogenic functions for membrane integration: the second transmembrane segment (TM2) is insufficient for the stop-transfer sequence, and TM3, TM5, and TM7 are not sufficient for the translocation initiation. In contrast to these phenomena, we herein demonstrate that TM2 shows an efficient stop-transfer function when it is near the preceding TM1 and suggest that TM3, TM5, and TM7 are followed by TM segments with a strong topogenic function to form Nexo/Ccyt topology, via which the preceding segments are integrated into the membrane. From these results, we propose that the interactions between the TMs should be operative during membrane integration, and that the segments with a weak topogenic function are given a transmembrane orientation by their following TMs.

Amino Acid Sequence↗

PubMed1998

Beta-casomorphin-5 stimulates neurite outgrowth in a mouse neuroblastoma cell line (Neuro-2a).

We studied the effect of beta-casomorphin-5 (mu-acting opioid peptides derived from milk protein beta-casein) on neurite outgrowth of mouse neuroblastoma cell line, Neuro-2a. Beta-casomorphin-5 stimulated neurite outgrowth of Neuro-2a cells in a naloxone-reversible manner. The stimulating effect of beta-casomorphin-5 was observed even at picomolar concentrations. The selective mu-agonist, (D-Ala2, N-Me-Phe4, Gly5-ol)-enkephalin (DAMGO) exhibited the similar stimulating effect only at micromolar concentrations. On the other hand, (D-Pen(2,5))-enkephalin (DPDPE) (a delta-agonist), U-50,488 (a kappa-agonist), and endogenous opioid peptides, such as enkephalins and dynorphin A (1-13), showed no such stimulating effect. These results suggest that the neurite outgrowth-stimulating action of beta-casomorphin-5 may be mediated via a receptor which has mu-like characteristics and high sensitivity to beta-casomorphin-5, and that beta-casomorphins may play a role as a neurite elongation factor during the suckling period.

Animals↗

PubMed1998

Characteristics of intracellular transferrin produced by human fibroblasts: its posttranscriptional regulation and association with tubulin.

Transferrin (Tf), an iron-binding protein, was investigated in the cultured human fibroblast which is a major cell type in connective tissues. Tf is a major iron-transporting protein and has an important role in iron metabolism. Using two-dimensional gel electrophoresis, we demonstrated the eight subtypes of Tf produced by cultured normal human fibroblasts and their down-regulation after the immortalization of human cells, an essential early step of in vitro transformation. However, the amount of Tf mRNA in the immortalized cells was equal to that in the normal human fibroblasts, suggesting that the down-regulation occurred at the posttranscriptional level. The amount of Tf receptor increased in the immortalized cells in spite of a decrease in the amount of intracellular Tf. Interestingly the produced Tf associated with microtubules. These findings suggest a novel aspect of Tf characteristics in human fibroblasts.

Cell Line↗

PubMed1998

Disruption of the RB pathway and cell-proliferative activity in non-small-cell lung cancers.

The pathway consisting of retinoblastoma protein (pRB), cyclin D1 and p16 (RB pathway) which is involved in the phosphorylation of pRB plays an important role in G1/S progression. The disruption of this RB pathway has been reported in several types of human neoplasm. An immunohistochemical study of 101 non-small-cell lung cancers (NSCLCs) showed loss of p16 is in 47 tumors (46.5%) and loss of pRB in 42 tumors (41.6%). In 79 of 101 NSCLCs (78.2%), the expression of p16 and pRB was complementary (p < 0.0001). Methylation of the cdkn2 gene was detected in 50% of p16-negative tumors and in 11% of p16-positive tumors. Aberrant expression of cyclin D1 was found in 45 tumors (44.5%). The cyclin-D1-positive tumors had significantly higher Ki-67 indices than the cyclin-D1-negative tumors irrespective of the tumor p16 or pRB expression. Thus, 91 (90%) of 101 NSCLCs showed disturbed expression of at least 1 of the 3 components of the RB pathway. Our results suggest that the disruption of the RB pathway plays an important role in tumorigenesis in NSCLCs and that increased cyclin-D1 expression leads to strong proliferative activity which may over-ride the suppressive effect of p16 and pRB.

Adenocarcinoma↗