Particulate matter air pollution exposure promotes recruitment of monocytes into atherosclerotic plaques.

Epidemiologic studies have shown an association between exposure to ambient particulate air pollution <10 microm in diameter (PM(10)) and increased cardiovascular morbidity and mortality. We previously showed that PM(10) exposure causes progression of atherosclerosis in coronary arteries. We postulate that the recruitment of monocytes from the circulation into atherosclerotic lesions is a key step in this PM(10)-induced acceleration of atherosclerosis. The study objective was to quantify the recruitment of circulating monocytes into vessel walls and the progression of atherosclerotic plaques induced by exposure to PM(10). Female Watanabe heritable hyperlipidemic rabbits, which naturally develop systemic atherosclerosis, were exposed to PM(10) (EHC-93) or vehicle by intratracheal instillation twice a week for 4 wk. Monocytes, labeled with 5-bromo-2'-deoxyuridine (BrdU) in donors, were transfused to recipient rabbits as whole blood, and the recruitment of BrdU-labeled cells into vessel walls and plaques in recipients was measured by quantitative histological methodology. Exposure to PM(10) caused progression of atherosclerotic lesions in thoracic and abdominal aorta. It also decreased circulating monocyte counts, decreased circulating monocytes expressing high levels of CD31 (platelet endothelial cell adhesion molecule-1) and CD49d (very late antigen-4 alpha-chain), and increased expression of CD54 (ICAM-1) and CD106 (VCAM-1) in plaques. Exposure to PM(10) increased the number of BrdU-labeled monocytes adherent to endothelium over plaques and increased the migration of BrdU-labeled monocytes into plaques and smooth muscle underneath plaques. We conclude that exposure to ambient air pollution particles promotes the recruitment of circulating monocytes into atherosclerotic plaques and speculate that this is a critically important step in the PM(10)-induced progression of atherosclerosis.

CCL23/myeloid progenitor inhibitory factor-1 inhibits production and release of polymorphonuclear leukocytes and monocytes from the bone marrow.

OBJECTIVE: CCL23/Myeloid progenitor inhibitory factor-1 is a human CC chemokine with potent in vitro suppressor effects on both human and murine myeloid progenitor cells. This study concerns in vivo inhibitory effect of CCL23 on production of polymorphonuclear leukocytes (PMNs) and monocytes in the bone marrow and their release into the circulation. METHODS: 5'-Bromo-2'-deoxyuridine (BrdU; 100 mg/kg) was used to label dividing PMNs and monocytes in the bone marrow, and BrdU-labeled cells were followed for 10 days in the circulation and identified using immunocytochemistry. Rabbits were given CCL23 (100 mug/kg, n = 5) or saline (control: n = 5) intravenously daily for 3 days before labeling with BrdU. Turnover of PMNs and monocytes in the bone marrow and their transit times through the bone marrow were calculated. RESULTS: CCL23 treatment tended to prolong transit time of PMN (98.4 +/- 4.3 hours vs 111.2 +/- 3.8 hours, control vs CCL23, p = 0.06) through the bone marrow and decreased the size of the bone marrow mitotic pool of PMN (p < 0.01). CCL23 treatment also prolonged the transit time of monocyte (43.4 +/- 3.1 hours vs 54.2 +/- 1.3 hours, control vs CCL23, p < 0.05) through the bone marrow and decreased turnover and pool size of monocytes in the bone marrow (p < 0.05). CONCLUSION: We conclude that CCL23 suppresses PMN and monocyte progenitors, decreases the pool size and slows their turnover in the bone marrow.

Expression of basic fibroblast growth factor is associated with a good outcome in patients with squamous cell carcinoma of the esophagus.

Angiogenesis is an essential step in tumor growth and metastasis, but rather than being controlled by means of a simple mechanism, the control of tumor angiogenesis may be mediated by several angiogenic factors. We investigated the expression of basic fibroblast growth factor (b-FGF) and platelet-derived endothelial cell growth factor (PD-ECGF) in squamous cell carcinoma of the esophagus in order to clarify the mechanism of angiogenesis. Expression of b-FGF and PD-ECGF was immunohistochemically investigated in tissue specimens from the tumors of 79 patients with squamous cell carcinoma of the esophagus who underwent curative esophagectomy without preoperative chemotherapy or radiation therapy, and the relationship between expression of b-FGF/PD-ECGF, microvessel density (MVD), and clinicopathological background factors was assessed. Tumor cells that expressed b-FGF were found in 41 patients (51.9%), and tumor cells that expressed PD-ECGF were found in 57 patients (72.2%). Although the mean vascular density (47.9/mm(2)) of b-FGF-positive tumors was significantly lower than that (67.2/mm(2)) of b-FGF-negative tumors (p=0.014), the difference between the 56.0/mm2 in PD-ECGF-positive tumors and 60.3/mm2 in PD-ECGF-negative tumors was not significant. Although the survival rate of patients with b-FGF-positive tumors was significantly higher than those with b-FGF-negative tumors (p=0.033), there was no significant difference between the survival rates of patients with PD-ECGF-positive and -negative tumors (p=0.580). Expression of b-FGF may be associated with promotion of angiogenesis and a good prognostic factor in squamous cell carcinoma of the esophagus.

Flow cytometric method for enumeration and characterization of newly released polymorphonuclear leukocytes from the bone marrow using 5'-bromo-2'-deoxyuridine.

Inflammation accelerates polymorphonuclear leukocyte (PMN) release from the bone marrow, and these PMNs are implicated in inappropriate tissue injury. We have previously developed a method using 5'-bromo-2'-deoxyuridine (BrdU) to study PMN kinetics using an immunocytochemical grading system of PMN on cytospin slides. The aim of this study was to develop a flow cytometric method to quantify the number of positively stained PMN and grade the intensity of staining for the transit time calculation of PMN through the marrow. Dividing myeloid progenitors in the marrow of rabbits were labeled with a pulse dosage of intravenous BrdU. BrdU-labeled PMN (PMN(BrdU)) were detected in the circulation using a FITC-conjugated anti-BrdU monoclonal antibody. The PMN(BrdU) were assigned to five groups according to their FITC intensity, and the transit times of PMN at different stages of development in the marrow were calculated. Results were compared using parallel immunocytochemical analysis of the same samples. In control animals, PMN(BrdU) in the circulation peaked at 72 h after BrdU labeling with 36.0% of PMN labeled. In normal rabbits, the transit times of PMN through the mitotic pool (49.5 +/- 4.2 h) and maturation pool (65.5 +/- 3.1 h) correlated well with immunocytochemical analysis and previously published values. Using this method, we demonstrated that exposure to air pollution particles accelerates the release of PMN(BrdU) from the marrow. We conclude that a flow cytometric approach for identifying BrdU-labeled leukocytes provides an objective and accurate method for studying leukocyte kinetics and behavior.

Particulate matter air pollution stimulates monocyte release from the bone marrow.

Particulate air pollution (PM(10)) stimulates alveolar macrophages (AMs) to release immature granulocytes from the bone marrow (BM) into the circulation. This study was designed to determine the effect of PM(10) (ambient EHC-93 or inert carbon [CC]) instillation exposure on the monocyte release from the BM and the role of AM in this response. Monocyte precursors were labeled in the BM of rabbits in vivo by an intravenous injection of 5'-bromo-2'-deoxyuridine, and the effects of PM(10) were determined by instillation either particles or supernatants of AM exposed to particles into the lungs. Instillation of EHC-93 (500 microg/ml) or supernatants from AM incubated with EHC-93 (100 microg/ml) increased circulating band cell counts (p < 0.05) and shortened the transit time of monocytes through the BM (35.5 +/- 2.2 to 25.0 +/- 1.5 hours or 36.2 +/- 2.6 to 25.7 +/- 1.8 hours, p < 0.05) compared with the control subject. CC (1%) instillation also shortened the monocyte BM transit time to 28.4 +/- 1.9 hours (p < 0.05), but supernatants of AM incubated with CC did not. We conclude that exposure to atmospheric PM(10) stimulates the production of mediators by AM, and these cytokines accelerate the monocyte release from the BM.

Exposure to ambient particles accelerates monocyte release from bone marrow in atherosclerotic rabbits.

Exposure to air pollution [particulate matter, particles <10 microm (PM(10))] causes a systemic inflammatory response that includes stimulation of the bone marrow (BM) and progression of atherosclerosis. Monocytes are known to play a key role in atherogenesis by migration into subendothelial lesions where they appear as foam cells. The present study was designed to quantify the BM monocyte response in Watanabe heritable hyperlipidemic (WHHL) rabbits after PM(10) exposure. WHHL rabbits were given twice weekly intrapharyngeal instillations of 5 mg of PM(10) for 4 wk to a total of 40 mg and compared with control WHHL or New Zealand White (NZW) rabbits. The thymidine analog 5'-bromo-2'-deoxyuridine was used to label dividing cells in the BM and a monoclonal antibody to identify monocytes in peripheral blood. The transit time of monocytes through the BM was faster in WHHL than in NZW rabbits (30.4 +/- 1.9 h vs. 35.2 +/- 0.9 h, WHHL vs. NZW; P < 0.05). PM(10) instillation exposure increased circulating band cell counts, caused rapid release of monocytes from the BM, and further shortened their transit time through the BM to 23.2 +/- 1.6 h (P < 0.05). The percentage of alveolar macrophages containing particles in the lung correlated with the BM transit time of monocytes (r(2) = 0.45, P <0.05). We conclude that atherosclerosis increases the release of monocytes from the BM, and PM(10) exposure accelerates this process in relation to the amount of particles phagocytosed by alveolar macrophages.

Loss of p16INK4a expression is associated with vascular endothelial growth factor expression in squamous cell carcinoma of the esophagus.

Vascular endothelial growth factor (VEGF) expression has been suggested to correlate with intratumoral microvessel density, tumor advancement and prognosis in esophageal squamous cell carcinoma (ESCC). Previous studies have showed that disruption of cell cycle regulator p16 is related to oncogenesis and tumor progression in ESCC. We hypothesized that VEGF expression in ESCC is reflected by abnormalities in the p16(INK4a) gene. To clarify the regulatory role of p16(INK4a) in VEGF expression in vitro, we transferred the p16(INK4a) gene into a p16(INK4a)-deleted ESCC cell line and observed changes in VEGF expression. Furthermore, we immunohistochemically assessed the expression of the cell cycle regulators (p16, p53 and RB) and VEGF in 90 surgically resected specimens of ESCC. Introduction of p16(INK4a) cDNA by the p16 expression vector significantly suppressed cell proliferation in the p16(INK4a)-deleted cell line TE8 (p < 0.0001). VEGF secretion by TE8 cells transfected with the p16(INK4a) vector was significantly suppressed as compared to non-transfected TE8 cells (p < 0.0001) and TE8 cells transfected with a control vector (p = 0.0015). The immunohistochemical studies of ESCC primary tumor specimens showed that loss of p16 expression was significantly correlated with VEGF-positive expression (p = 0.0004). The cumulative postoperative survival rate in the group with p16-positive and VEGF-negative expression was significantly higher than in the other groups. Neither p53 nor RB expression had any impact on outcome. Aberrant p53 expression tended to be associated with VEGF expression, but the trend did not reach statistical significance. Our study demonstrated that VEGF expression was correlated with p16 expression in ESCC. Our results suggest that p16 may have a regulatory role in VEGF expression in ESCC.

Monocyte recruitment into the lungs in pneumococcal pneumonia.

The recruitment of monocytes into the alveolar spaces is crucial for clearing infections and resolving the inflammatory response. We have previously reported the effect of acute pneumonia on monocyte transport through the bone marrow, and the present study concerns their clearance from the blood and migration into the lung airspaces. Dividing monocytes were labeled with the thymidine analog, 5'-bromo-2'-deoxyuridine (BrdU). Whole blood containing the labeled monocytes (MO(BrdU)) was transfused from either donor rabbits with pneumonia or from uninfected controls into recipients with pneumonia, where they were detected in blood and tissues using a double immunostaining method. The results show that MO(BrdU) from infected animals rapidly disappeared from the circulation (P < 0.05), preferentially sequestered in the infected lung tissue within 1 h (22.0 +/- 3.3% versus 6.0 +/- 0.4%, pneumonic region versus peripheral blood, P < 0.05), and accumulated to a greater degree in pneumonic airspaces than control monocytes 48 h after transfusion (3.9 +/- 0.5% versus 1.1 +/- 0.1%, P < 0.05). We conclude that immature monocytes released from the marrow by pneumonia sequester rapidly in lung microvessels but their migration in pneumonic airspaces is delayed.

[Chemotherapy for the patients with esophageal cancer].

The combination of cisplatin and continuous-infusion 5-fluorouracil is the standard regimen for the treatment of both squamous cell carcinoma and adenocarcinoma. Paclitaxel has shown favorable results as a single agent or in combination with cisplatin. The efficacy of neoadjuvant chemotherapy in terms of survival benefit remains controversial despite large-scale, randomized, controlled trials comparing it with surgery alone. The disease-free survival benefit of postoperative adjuvant chemotherapy was recognized in a Japan Clinical Oncology Group randomized controlled trial in comparison with surgery alone.