Tetsuji Yamashita

Increased levels of vascular endothelial growth factor and interleukin-6 in the aqueous humor of diabetics with macular edema

PURPOSE To investigate the relationship between diabetic macular edema and the levels of vascular endothelial growth factor (VEGF) and interleukin-6 (IL-6) in aqueous humor and plasma. DESIGN Comparative cross-sectional study. METHODS Fifty-four eyes from 54 diabetic patients were used. The concentrations of VEGF and IL-6 in undiluted aqueous specimens (obtained from the eyes during cataract surgery) and in plasma were measured by an enzyme-linked immunosorbent assay. To assess blood-aqueous barrier function, the aqueous flare intensity was measured by a laser flare-cell meter as an estimate of the aqueous protein level. RESULTS The aqueous levels of VEGF and IL-6 were significantly correlated with the severity of macular edema (rho = 0.628, P <.001 and rho = 0.517, P <.01, respectively), as well as with the aqueous protein concentration (rho = 0.618, P <.001 and rho = 0.588, P <.001, respectively). Aqueous levels of VEGF and IL-6 were significantly higher than their respective plasma levels (both P <.001). In addition, the aqueous level of VEGF was significantly correlated with that of IL-6 (rho = 0.537, P <.01). Furthermore, the status of the posterior vitreous significantly correlated with the severity of macular edema (rho = 0.618, P <.0001). CONCLUSIONS These results suggest that both VEGF and IL-6 are produced together in the intraocular tissues, and are involved in the pathogenesis of macular edema.

IL-9 Enhances the Growth of Human Mast Cell Progenitors Under Stimulation with Stem Cell Factor

We examined the effects of IL-9 on human mast cell development from CD34+ cord blood (CB) and peripheral blood cells in serum-deprived cultures. IL-9 apparently enhanced cell production under stimulation with stem cell factor (SCF) from CD34+ CB cells. A great majority of the cultured cells grown with SCF + IL-9 became positive for tryptase at 4 wk. In methylcellulose cultures of CD34+ CB cells, IL-9 increased both the number and size of mast cell colonies grown with SCF. Furthermore, SCF + IL-9 caused an exclusive expansion of mast cell colony-forming cells in a 2-wk liquid culture of CD34+ CB cells, at a level markedly greater than for SCF alone. Clonal cell cultures and RT-PCR analysis showed that the targets of SCF + IL-9 were the CD34+CD38+ CB cells rather than the CD34+CD38− CB cells. IL-9 neither augmented the SCF-dependent generation of progeny nor supported the survival of 6-wk-cultured mast cells. Moreover, there was no difference in the appearance of tryptase+ cells and histamine content in the cultured cells between SCF and SCF + IL-9. The addition of IL-9 increased numbers of mast cell colonies grown with SCF from CD34+ peripheral blood cells in children with or without asthma. It is of interest that mast cell progenitors of asthmatic patients responded to SCF + IL-9 to a greater extent than those of normal controls. Taken together, IL-9 appears to act as a potent enhancer for the SCF-dependent growth of mast cell progenitors in humans, particularly asthmatic patients.

Circulating vascular endothelial growth factor (VEGF) is a possible tumor marker for metastasis in human hepatocellular carcinoma

Abstract: Vascular endothelial growth factor (VEGF) is closely related to angiogenesis in various human cancers. However, little is known of its circulating levels in hepatocellular carcinoma (HCC). We examined circulating VEGF levels in chronic liver disease to assess their clinical significance. Plasma VEGF concentrations were determined, by enzyme immunoassay, in patients with chronic hepatitis (CH; n = 36), liver cirrhosis (LC; n = 77), and HCC (n = 86) for a cross-sectional study. Plasma VEGF levels in healthy controls (n = 20) and CH, LC, and HCC patients were 17.7 ± 5.4 (mean ± SD), 30.6 ± 22.8, 34.4 ± 27.0, and 51.1 ± 71.9 pg/ml, respectively. The levels were significantly elevated in the HCC group, compared with the control, CH, and LC groups. Plasma VEGF levels in stage I, II, III, IVA, and IVB HCC patients were 27.6 ± 16.1, 26.5 ± 13.7, 35.8 ± 15.3, 45.4 ± 39.4, and 103.1 ± 123.2 pg/ml, respectively. The stage IVB patients with remote metastasis showed significantly marked elevation compared with the patients at the other stages. Platelet numbers were weakly correlated with plasma VEGF levels in the HCC group. Plasma VEGF level was highly elevated in patients with HCC, particularly those with metastatic disease. We consider that plasma VEGF is a possible tumor marker for metastasis of HCC. Circulating VEGF may be derived mainly from the large burden of tumor cells, and partly from platelets activated by the vascular invasion of HCC cells.

New bronchoscopic microsample probe to measure the biochemical constituents in epithelial lining fluid of patients with acute respiratory distress syndrome

ObjectiveA noninvasive bronchoscopic microsampling (BMS) probe was developed to sample biochemical constituents of the epithelial lining fluid in small airways. DesignObservational, controlled study. SettingIntensive care unit of academic medical center. Patients and Procedure BMS was applied in a control group of seven patients who had hemoptysis or small solitary peripheral nodules but no hypoxemia or other signs of acute inflammation and in four patients with acute respiratory distress syndrome (ARDS), to test whether BMS can ascertain the presence of acute pulmonary inflammation without complications. Measurements and Results Complications, including a significant decrease in arterial oxygen saturation, were observed neither during nor after BMS. In the ARDS group, albumin, lactate dehydrogenase, interleukin-6, basic fibroblast growth factor, and neutrophil elastase concentrations in epithelial lining fluid were significantly higher (p < .0001, p = .012, p < .0001, p < .0001, and p < .0001, respectively) than in the control group. Serial BMS was safely performed in one patient with ARDS, allowing us to observe a correlation between changes in the concentration of inflammation-related biochemical markers and clinical course of the disease. ConclusionsThese results suggest that BMS is safe and useful to monitor pulmonary biochemical events in ARDS.

Expression of Histamine Receptors in Nasal Epithelial Cells and Endothelial Cells – The Effects of Sex Hormones

Background: Hyperreactivity of the nasal mucosa is a characteristic of nasal allergy. During pregnancy, aggravation of nasal allergic symptoms is occasionally observed in subjects with nasal allergy. Methods: Using the reverse transcription-polymerase chain reaction and Southern blot hybridization method, we investigated histamine H1 receptor mRNA (H1R mRNA) expressions in specimens of nasal epithelial layer obtained by scraping, as well as cultured human nasal epithelial cells (HNECs) and human mucosal microvascular endothelial cells (HMMECs). We compared the expressions on the specimens from patients with nasal allergy with those with nonallergic rhinitis or those from normal volunteers. In addition, we investigated the effects of female hormones on the H1R mRNA expressions in HNECs and HMMECs. Results: H1R mRNA was detected in scraped specimens of nasal epithelial layer, as well as in HNECs and HMMECs. The mRNA expressions in nasal mucosal scraped specimens of epithelial layers and HNECs were more marked in patients with nasal allergy than in the other two groups. In addition, the present study demonstrates that the female hormones β-estradiol and progesterone significantly increase the expressions of H1R mRNA on HNECs and HMMECs. Conclusion: The increase of the expressions of H1R mRNA may explain, in part, the nasal hyperreactivity to histamine observed in patients with nasal allergy. It has also been suggested that sex hormones are related to the preponderance of females in the incidence of allergic rhinitis after puberty, and that they are related, at least partially, to the aggravation of the nasal hyperreactivity symptoms during pregnancy through the enhanced expression of H1R mRNA on HNECs and HMMECs.

Effect of Female Hormones on the Production of IL-4 and IL-13 from Peripheral Blood Mononuclear Cells

In this study we compared the concentrations of IL-4, IL-13, and IFN-gamma, which were produced by human peripheral blood mononuclear cells (PBMC) in the presence or absence of preincubation with beta-estradiol or progesterone both after a specific antigen challenge and without a specific antigen challenge. The concentrations of IL-4 and IL-13 from PBMC which had been preincubated with progesterone or gamma-estradiol for 18-24 h were significantly greater than those of IL-4 and IL-13 from PBMC which had been preincubated with PBS, the control. On the other hand, the concentration of IFN-gamma from PBMC was unchanged. We were able to confirm that the female hormones beta-estradiol and progesterone, at levels similar to those occurring during pregnancy, have the ability to induce production of IL-4 and IL-13 in human mononuclear cells. These results suggest that female hormones may aggravate nasal allergy symptoms during pregnancy by increasing IgE synthesis and inducing selective eosinophil infiltration.

IL-4 upregulates FcϵRI α-chain messenger RNA in eosinophils

Abstract By using the reverse transcription polymerase chain reaction and Southern blot hybridization, we demonstrated that FcϵRI α-chain (FcϵRIα) messenger RNA was expressed in eosinophils purified from the peripheral blood of patients with allergic rhinitis and that this expression was enhanced by IL-4. However, studies in which flow cytometry or immunostaining was used did not reveal the expression of FcϵRIα protein on eosinophils from peripheral blood. Neither IL-4 alone nor the combination of IL-4 and other cytokines could induce detectable FcϵRIα protein; nevertheless, they do express FcϵRIα mRNA. Double-labeling immunostaining on cryostat sections of nasal mucosa clearly demonstrated that some FcϵRIα-positive cells were eosinophil cationic protein-positive, which confirms their eosinophilic nature. Not all the eosinophil cationic protein-positive cells express an FcϵRIα signal. Considering that FcϵRIα mRNA was detectable in four of five samples of eosinophils from those patients with nasal allergy and that only one of five eosinophil samples from normal subjects expressed FcϵRIα mRNA, the level of FcϵRI expression may be correlated with the activation of eosinophils. It seems very likely that some other unidentified factors are required for the process from the expression of FcϵRIα mRNA to that of FcϵRI as a protein. (J A LLERGY C LIN I MMUNOL 1995;96:1161-9.)

Elevation of the plasma level of RANTES during asthma attacks

BACKGROUND RANTES is considered to play an important role in various immune and allergic disorders since it is a potent chemoattractant for inflammatory cells such as eosinophils, memory T cells, and monocytes. OBJECTIVE To investigate the possible role of RANTES in the pathogenesis of bronchial asthma. METHODS The plasma level of RANTES was measured in 12 asthma patients and 15 normal controls by a sandwich enzyme-linked immunosorbent assay. RESULTS In the patients with asthma, the plasma RANTES level was significantly elevated during acute attacks compared to that in the controls. In addition, it was higher than that during the asymptomatic state in the same patients. Plasma beta-thromboglobulin levels were also elevated in asthma patients during acute attacks and showed a correlation with the RANTES level. CONCLUSION These findings suggest a role for RANTES in the pathogenesis of asthma and a possible role for platelets in RANTES release in asthma.

Plasma level of basic fibroblast growth factor increases with progression of chronic liver disease

Basic fibroblast growth factor (FGF) is thought to be involved in carcinogenesis and, to clarify its clinical significance, the study of its blood level in cancer patients is important. Plasma levels of basic FGF are reported to be elevated in some cancers. However, little is known of basic FGF levels in plasma in hepatocellular carcinoma (HCC). In this study, we measured basic FGF plasma levels in patients with chronic liver disease and compared the levels in chronic hepatitis (CH), liver cirrhosis (LC), and HCC. We also examined whether these levels were related to serum levels of asparate aminotransferase, alanine aminotransferase, γ-glutamyl transpeptidase, alkaline phosphatase, leucine aminopeptidase, total bilirubin, total protein, and albumin, and to the indocyanine green test (i.e., liver function tests) and to type III procollagen, 7S domain of IV type collagen, and hyaluronic acid (i.e., markers of liver fibrosis). Levels of basic FGF. determined by a quantitative “sandwich” enzyme immunoassay, were significantly elevated with the progression of liver disease; being 3.67 ± 2.37 (mean ± SD), 7.78 ± 6.61, and 12.37 ± 7.67pg/ml in the CH, LC, and HCC groups, respectively. FGF levels were elevated to a greater extent in the HCC patients than in the CH (P<0.0001) and LC patients (P=0.0117). Levels were higher in LC than in CH (P=0.0204). None of the liver function test findings or levels of markers of liver fibrosis were correlated with levels of basic FGF. These results suggest that circulating basic FGF could serve as a new indicator of the progression of chronic liver disease. The extremely elevated plasma of level basic FGF in the HCC group suggests that basic FGF may be related to the development of HCC.

Stimulation and Inhibition of Angiogenesis in Diabetic Retinopathy

PURPOSE To investigate the role of stimulators and inhibitors of angiogenesis in the pathogenesis of diabetic retinopathy. METHODS Undiluted vitreous samples and simultaneous paired plasma samples were obtained from 30 diabetic patients (35 eyes) undergoing vitreous surgery. The levels of vascular endothelial growth factor (VEGF), endostatin, and platelet factor-4 (PF-4) were measured simultaneously in each specimen by enzyme-linked immunosorbent assay. The severity of diabetic retinopathy was evaluated according to the modified Early Treatment Diabetic Retinopathy Study retinopathy severity scale. RESULTS Vitreous levels of VEGF and endostatin were significantly correlated with the severity of diabetic retinopathy (rho = 0.52, rho = 0.48, respectively), but the vitreous level of PF-4 was not (rho = 0.12). Vitreous levels of VEGF, endostatin, and PF-4 were not significantly correlated with their plasma levels. The vitreous level of VEGF was significantly correlated with that of endostatin (rho = 0.42). The VEGF concentration was significantly higher in the vitreous than in the plasma, while the endostatin concentration was not. CONCLUSIONS The present study showed that VEGF and endostatin were expressed in the vitreous of patients with diabetic retinopathy and may be involved in the pathogenesis of this condition.