Katsunori Fukuda

Type V collagen selectively inhibits human endothelial cell proliferation.

Type V collagen from human placenta remarkably inhibited human umbilical vein endothelial cell (HUVEC) proliferation in a dose-dependent manner when coated on the culture dishes. Other types of collagen (I, III, IV) and fibronectin enhanced HUVEC proliferation under the same conditions. The inhibitory activity of type V collagen was seen not only when it was coated on the dishes, but also when it was directly added into cell culture. The attachment effect of type V collagen did not differ from that of type I collagen. The inhibitory activity is a phenomenon selective for endothelial cells, since type V collagen did not affect the proliferation of human umbilical vein smooth muscle cells, aortic smooth muscle cells, or nasal mucosa fibroblasts.

Th1/Th2 response profiles to the major allergens Cry j 1 and Cry j 2 of Japanese cedar pollen.

Cry j 1 and Cry j 2 are known to be the major allergens of Japanese cedar pollen. A comparative study was carried out on the immune responses to stimulation with Cry j 1 and Cry j 2 in 24 symptomatic patients and six nonallergic subjects. In T‐cell proliferation assays, mean stimulation indexes (SI) were 10.6 for Cry j 1 and 11.7 for Cry j 2 stimulation, respectively, in the allergic patients. Two of the nonallergic subjects showed strong T‐cell proliferation to both allergens, while the remainder did not. All the allergic subjects (17/17) showed high titers of anti‐Cry j 1 IgE antibody at a mean value of 165 U/ml, whereas only 64% responded to Cry j 2 with low titers at a mean value of 26 U/ml. Nonallergic subjects did not respond with IgE production. Allergic subjects were further examined for their cytokine production profiles. All allergic subjects tested (16/16) produced high levels of interferon‐gamma (IFN‐γ) in response to Cry j 1 with a mean value of 918 pg/ml, while only five subjects showed significant elevation of IFN‐γ production in response to Cry j 2 with a mean value of 679 pg/ml. The remainder produced small amounts of IFN‐γ. Cry j 1 induced higher levels of interleukin (1L)‐10 gene expression than did Cry j 2 stimulation, while both allergens induced IL‐4 expression at a similar level. The IL‐12 p35 gene was constitutively expressed, whereas the IL‐12 p40 gene expression in Cry j 1‐stimulated cells was elevated eightfold over that of nonstimulated cells. Increased expression of the IL‐12 p40 gene was negligible in Cry j 2‐stimulated cells. Thus, Cry j 1 stimulated mixed features of Th1 and Th2‐like responses, while Cry j 2 played a minor role in inducing IgE production and cytokine (IFN‐γ, IL‐10, and IL‐12) production, except for IL‐2 production and strong T‐cell proliferative activity. Therefore, it was concluded that Cry j 1 is the more important allergen, and that T‐cell proliferation assays do not necessarily reflect the level of allergenicity.

Localization of il‐1β mrna and cell adhesion molecules in the maxillary sinus mucosa of patients with chronic sinusitis

Interleukin‐1β (IL‐1β) is a predominant cytokine in retained paranasal sinus fluid of chronic sinusitis where infiltration by polymorphonuclear neutrophils (PMNs) of nasal and paranasal mucosa is characteristic. The authors investigated the localization of IL‐1β messenger RNA (mRNA) in the maxillary sinus mucosa of patients with chronic sinusitis, using digoxigenin‐labeled oligonucleotide probes. IL‐1β mRNA was detected in some extravascular PMNs and small numbers of mononuclear leukocytes but was not detected in other tissue cells or intravascular leukocytes. The expression and distribution of the cell adhesion molecules, intercellular adhesion molecule‐1 (ICAM‐1) and endothelial‐leukocyte adhesion molecule‐1 (ELAM‐1), were also studied in cultured human mucosal microvascular endothelial cells and in the maxillary sinus mucosa in chronic sinusitis by immunohistochemistry using monoclonal antibodies against these cell adhesion molecules. Only ICAM‐1 was expressed on cultured human mucosal microvascular endothelial cells without IL‐1β stimulation. With IL‐1β activation of these cells, ELAM‐1 was expressed strongly and the expression of ICAM‐1 was enhanced. In the maxillary sinus mucosa, ICAM‐1 was strongly and universally expressed on endothelial cells of all small vessels, whereas ELAM‐1 was expressed only in the subepithelial region. These findings suggest that IL‐1β, one of mediators in chronic sinusitis, is produced by PMNs, induces the expression of ICAM‐1 and ELAM‐1 on endothelial cells, and, thereby, stimulates PMN infiltration in chronic sinusitis.

Establishment of human mucosal microvascular endothelial cells from inferior turbinate in culture

Human microvascular endothelial cells were isolated and cultured from the mucosa of inferior turbinates. Using dish-coated collagen and a medium composed of a 1:1 mixture of sarcoma 180-conditioned medium and Dulbeccos modified Eagles medium (containing 10% fetal bovine serum and 75 micrograms/mL endothelial cell growth factors prepared from bovine pituitary glands), these cells grew rapidly to confluence and survived serial passages until the 16th population doubling level. The cells were identified as endothelial cells by their morphology, immunostaining of factor VIII antigen, and cytochemical staining with Ulex europeus agglutinin. Furthermore, Weibel-Palade bodies and numerous pinocytotic vesicles were confirmed by electron microscopy. Proliferation experiments demonstrated the need for either endothelial cell growth factor or tumor-conditioned medium. An exogenous matrix was also required for these cells in tissue culture. A tubule-like morphology appeared in the original monolayer of human microvascular endothelial cells after 1 month in the same plate, indicating that these cells have the ability to form tubules in the presence of sarcoma 180-conditioned medium.

Immunodominance of seven regions of a major allergen, Cry j 2, of Japanese cedar pollen for T-cell immunity

The immunodominant regions of the Japanese cedar pollen allergen Cry j 2 for T‐cell immunity were determined with whole peripheral blood lymphocytes (PBL) derived from seven allergic patients and three nonallergic subjects. Cry j 2‐stimulated T‐cell proliferation was inhibited by anti‐HLA‐DR. but not by anti‐HLA‐DQ antibody, indicating that the responding T cells recognized the allergen peptides associated with HLA‐DR molecules. It was found that seven regions of Cry j 2, i.e., regions corresponding to amino acid numbers 1–26, 70–84, 151–167. 187–203, 252–279, 283–314, and 345–362, were immunodominant for T‐cell proliferation. Thus, Cry j 2 bears a limited number of immunodominant regions despite polymorphic features of HLA‐DR in the immune system. This suggests the possibility of molecularly designing Cry j 2 antagonists that could downregulate allergic reactions to Japanese cedar pollen.

Synovial sarcoma in the parapharyngeal space: case report and review of the literature.

We encountered a rare case of synovial sarcoma in the parapharynx of a 47-year-old Japanese man. This patient presented with an enlarging tumor in the right side of his neck that had grown progressively over a 4-week period. Radiological examinations revealed that the tumor arose from the parapharyngeal space. The tumor could not be completely removed at surgery. Metastasis to the lumbar vertebra was detected postoperatively. The patient underwent three courses of chemotherapy and the delivery of palliative radiation to the lumbar vertebra without success. The patient died of lung metastasis 7 months after surgery.

Retention fluids of chronic sinusitis induce neutrophil adherence to microvascular endothelial cells

The adherence of circulating leukocytes to the vascular endothelium is a critical step in the emigration of leukocytes through blood vessel walls to inflammatory lesions. The influence of nasal secretions on the adherence of neutrophils to the vascular endothelium was investigated using monolayers of human mucosal microvascular endothelial cells derived from the inferior turbinate. Preincubation of vascular endothelial cells with retention fluids from the maxillary sinus of the patients with chronic sinusitis showed increased neutrophil adherence. Recombinant IL-1 beta was also tested and found to induce adherence of neutrophils to human mucosal microvascular endothelial cells. However, no adhesive effect was observed with the nasal secretions of nasal allergy. An enzyme-linked immunosorbent assay detected considerable amounts of IL-1 beta in the chronic sinusitis retention fluids, while the amounts of IL-1 alpha and TNF-alpha were very low. The increased adhesion of the neutrophils by the retention fluids of chronic sinusitis was also neutralized by the incubation with anti-IL-1 beta antibody in a dose dependent manner. These findings suggest that IL-1 beta in the paranasal secretion of chronic sinusitis induces the adherence of neutrophils to vascular endothelium and subsequent infiltration of neutrophils in the paranasal sinuses, thus contributing to the persistence of chronic sinusitis.

A study on the relationship between adenoid vegetation and rhinosinusitis

The relationship between adenoidal-nasopharyngeal ratios (AN ratios) and sinusitis were evaluated in 404 children. The AN ratios of children with snoring, mouth breathing, or nasal obstruction (major symptoms of adenoidal enlargement) were significantly higher than those of children without these symptoms. However, the AN ratios of children with sinusitis were almost equal to those of normal children.

Effects of oxygen radicals on ciliary motility in cultured human respiratory epithelial cells.

There are few reports about direct effects of specific oxygen products on ciliary function because of their instability and reactivity. We investigated the direct effects of superoxide anion (O2-) and of hydrogen peroxide (H2O2) on the ciliary function of human respiratory epithelial cells, using monolayer cell cultures, high speed video analysis of frequency (CBF), amplitude (CBA), and coordination of ciliary beats and evaluating the surface structural changes of ciliated cells at the same time. 10(-2) M H2O2 decreased ciliary beat activity. The CBF was 36.5 +/- 4.4% and the CBA was 51.0 +/- 3.8% of the baseline (time = 0) after 5 min (all p < 0.001). Catalase (2 micrograms/ml) abolished the ciliotoxic effect of H2O2. The O2- produced by reaction of xanthine (0.06 mM)-xanthine oxidase (0.04 U/ml) caused a temporary rapid increase of 26.8 +/- 1.7% in CBF and an increase of 42.5 +/- 4.1% in CBA after 15 sec (all p < 0.001). Superoxide dismutase significantly reduced these increases. Results indicated that O2- activated ciliary function with a temporary increase in O2(-)-production. This suggests that the removal of H2O2 from the O2- reaction is important in improving mucociliary clearance in excessive oxygen metabolites.

Chondrosarcoma of the hyoid bone.

Chondrosarcoma of the head and neck is relatively rare. We report the sixth case known to us in which the chondrosarcoma had originated in the hyoid bone. The patient, a 66-year-old man, underwent surgery. The resected tumour was a 5 cm encapsulated lesion composed of lobulated, firm, gray-white tissue. Histologically it showed mature chondrocytes without capsular invasion. There was high cellularity and a few binucleated chondrocytes. Although the tumour grade was prognostic significance, our literature review of the five other cases suggests that the location of the primary lesion and the adequacy of its resection to be more important. Our patient continues to do well 15 months post-operatively. However, long-term follow-up is essential as recurrences have been reported even after eight or 10 years.