Hypoxic induction of interleukin-8 gene expression in human endothelial cells.

Because leukocyte-mediated tissue damage is an important component of the pathologic picture in ischemia/reperfusion, we have sought mechanisms by which PMNs are directed into hypoxic tissue. Incubation of human endothelial cells (ECs) in hypoxia, PO2 approximately 14-18 Torr, led to time-dependent release of IL-8 antigen into the conditioned medium; this was accompanied by increased chemotactic activity for PMNs, blocked by antibody to IL-8. Production of IL-8 by hypoxic ECs occurred concomitantly with both increased levels of IL-8 mRNA, based on polymerase chain reaction analysis, and increased IL-8 transcription, based on nuclear run-on assays. Northern analysis of mRNA from hypoxic ECs also demonstrated increased levels of mRNA for macrophage chemotactic protein-1, another member of the chemokine superfamily of proinflammatory cytokines. IL-8 gene induction was associated with the presence of increased binding activity in nuclear extracts from hypoxic ECs for the NF-kB site. Studies with human umbilical vein segments exposed to hypoxia also demonstrated increased elaboration of IL-8 antigen compared with normoxic controls. In mice exposed to hypoxia (PO2 approximately 30-40 Torr), there was increased pulmonary leukostasis, as evidenced by increased myeloperoxidase activity in tissue homogenates. In parallel, increased levels of transcripts for IP-10, a murine homologue in the chemokine family related to IL-8, were observed in hypoxic lung tissue. Taken together, these data suggest that hypoxia constitutes a stimulus for leukocyte chemotaxis and tissue leukostasis.

Pre-operative assessment of cancer in the elderly (PACE): a comprehensive assessment of underlying characteristics of elderly cancer patients prior to elective surgery.

BACKGROUND: Cancer is a disease that particularly affects the elderly and, although surgery is the first treatment choice, many elderly cancer patients do not receive standard surgery because they are considered unfit for treatment due to an inaccurate estimation of operative risk. Pre-operative Assessment of Cancer in the Elderly (PACE) was developed in order to address the need to provide detailed information about the functional reserve of the elderly cancer patient to aid individualised management. METHODS: PACE incorporates a battery of validated instruments including the Comprehensive Geriatric Assessment (CGA), Brief Fatigue Inventory (BFI), Eastern Cooperative Oncology Group Performance Status (ECOG-PS), and American Society Anesthesiologists (ASA) grade. An international prospective study was conducted with 460 consecutive elderly cancer patients (216 breast, 146 GIT, 71 GUT, 27 other) receiving PACE prior to receiving elective surgery. RESULTS: Three hundred and eighty four patients (83.4%) were observed to have at least one co-morbidity; the most common being hypertension (n=246, 53.5%). More than two thirds of the patients had good functional and mental status according to PACE. After adjusting for age, sex and type of cancer, six of the seven items of PACE were found to be significantly associated with co-morbidities (according to the Satariano's Index of Co-morbidities (SIC)). A multivariate analysis identified IADL, BFI and ASA to be the most important instruments in explaining SIC. DISCUSSION: PACE has been effectively used to describe the functional capacity and health status in an international cohort of elderly cancer patients. The majority of PACE instruments have been found to be significantly associated with co-morbidities (SIC) and can distinguish between type and severity of cancer. PACE represents a useful tool in evaluating onco-geriatric fitness for surgery.

[Bronchial reactivity to methacholine and serotonin in six inbred mouse strains].

We examined six mouse strains, A/J, DBA/2, WBB6F1-W/Wv, WBB6F1- +/+, C3H/HeN, C57BL/6, for their bronchial reactivity to methacholine and serotonin. The mice were anaesthetized with intraperitoneal sodium pentobarbital. The trachea was intubated and one jugular vein was cannulated. Then, the mice were ventilated on air, using a small animal ventilator at a rate of 60 strokes/min with a tidal volume of 0.5 ml. Methacholine and serotonin diluted to various concentrations were administered intravenously. Bronchoconstriction was assessed by the modified Konzett-Rossler method. According to their bronchial responsiveness to methacholine and serotonin, the strains were grouped into high- (A/J, DBA/2), middle- (WBB6F1-W/Wv, -+/+) or low- (C3H/HeN, C57BL/6) responder groups.

[Endoscopic treatment of colorectal cancer].

Endoscopic polypectomy should be applied only for early colorectal carcinomas. Intramucosal carcinoma do not have a risk of lymph node metastases. However, there is an about 10% risk of lymph node metastases among carcinomas showing submucosal invasion (sm carcinoma). When risk factors revealed to be positive after polypectomy, subsequent surgical resection of the large bowel with lymph nodes dissection is needed, because these sm carcinomas are considered to have a high risk of lymph node metastases. Therefore, accurate diagnosis of depth of invasion is essential to prevent subsequent surgical resection following endoscopic polypectomy. Endoscopy, barium enema and endoscopic ultrasonography (EUS) are all considered to be effective for an accurate diagnosis of depth of invasion. Endoscopic polypectomy includes hot biopsy, snare polypectomy and endoscopic mucosal resection (EMR). Appropriate maneuver must be chosen, considering the characteristics of the lesion. Major complications after endoscopic polypectomy are bleeding and perforation of the large bowel. Including an establishment of a new risk factors, further efforts must be made to prevent unnecessary additional surgical resection of the large bowel following endoscopic polypectomy.

[Ozone-induced airway hyperresponsiveness in guinea pigs].

We studied the time related changes of airway hyperresponsiveness induced by ozone inhalation (2.9 ppm, 30 min) in guinea pigs. In unanesthetized and spontaneously breathing guinea pigs, the respiratory resistance was measured by a forced oscillation technique. The respiratory resistance and respiratory frequency were unchanged until 24 hours after ozone inhalation. In mechanically ventilated guinea pigs, airway responsiveness to inhaled methacholine was determined using a modification of the Konzett-Rössler technique, and after methacholine challenge bronchoalveolar lavage (BAL) was performed for cell yield. At 1 hour and 3 hours after ozone inhalation, airway responsiveness was increased significantly, but returned to the control level at 6 hours. In the BAL fluid, there was a significant increase in neutrophils at 3 hours after ozone inhalation and thereafter. In the separated groups, before air or ozone inhalation, human serum albumin (HSA) was administered intravenously, and BAL was performed 1 hour after inhalation. In the ozone inhalation group, the concentration of HSA in BALF was increased significantly compared to the air inhalation group. These results suggest that airway hyperresponsiveness induced by ozone inhalation may occur before the influx of neutrophils into the airways and may depend on some structural changes such as submucosal and mucosal edema induced by the enhancement of capillary permeability.

[Feasibility of DNA analysis for prognostic evaluations of colorectal cancer using touch preparations and image cytometry].

The purpose of this study is to demonstrate the applicability of DNA ploidy analysis by using touch preparations obtained from fresh and frozen tissues on the prognostic evaluations of colorectal cancer, especially the development of hepatic metastases. The specimens obtained from surgically resected colorectal carcinomas (32), and hepatic metastases (4), were lightly touched with the slide glasses. Among the 32 patients with colorectal cancers, 7 had synchronous liver metastases, and 5 metastases developed metachronously. After formalin-fixation and Feulgen staining, the DNA analysis was performed by image cytometer (ICM, CAS200R). The data were expressed as the ploidy patterns and DNA indices (p-DNAI: peak DNA index, m-DNAI: mean DNA index), and were correlated very well with those obtained from cytophotometry. The DNA ploidy pattern among the 32 colorectal cancer specimens were Aneuploid(A) in 16, Polyploid (P) in 7, Diploid (D) in 4 and D+P in 5. The cases with synchronous or metachronous liver metastases had a higher m-DNAI (1.74 +/- 0.28) and were Aneuploid in 8, P in 3, and D+P in 1 case. Aneuploidy or Polyploidy predominated among the cases with metachronous liver metastases (4 in 5 cases), with m-DNAI higher than 1.5. Thus, "m-DNA > 1.5" could be a useful prognostic indicator of the high risk group of liver metastases. The liver metastases specimens had Aneuploid and Polyploid patterns in 2 cases each, with m-DNAI ranging from 1.88 to 2.31. DNA ploidy pattern analysis using touch preparations with image cytometry may be useful in determining the prognosis for colorectal cancer.

[The changes in cell population and chemical mediators in BALF following local allergen challenge].

In eight subjects who showed dual asthmatic response (DAR) in bronchial provocation tests (BPT) with specific allergens, local allergen challenge (LAC) was conducted using a flexible bronchofiberscope. We examined the concentrations of histamine, leukotriene B4 and C4 (LTB4 and LTC4) and cell populations in bronchoalveolar lavage fluid (BALF) collected before LAC (control), during the immediate response phase (IR), and the late response phase (LR). Control BALF was collected from the left lingula (B4 or B5), and BALF in the IR or LR phase from the right middle lobe (B4 or B5). Each BAL was conducted with 50 ml of saline at 37 degrees C and repeated three times in succession. It was noted that histamine increased significantly (p less than 0.05) in IR-BALF from the control level. In addition, the concentrations of LTC4 and the numbers of eosinophils increased in IR-BALF. In LR-BALF, the numbers of eosinophils (p less than 0.01), and the concentrations of histamine (p less than 0.05), LTC4 (p less than 0.05) and LTB4 increased. From these results, it was suggested that infiltration of eosinophils and various chemical mediators in the bronchial mucosa play important roles in the development of late asthmatic response.

[The mechanism of airway hyperresponsiveness following immediate bronchial response in ovalbumin (OA)-sensitized guinea pigs].

We have previously demonstrated that airway responsiveness was enhanced following a late bronchial response (LBR) after an allergen challenge in ovalbumin (OA)-sensitized guinea pigs. The purpose of the present studies was to evaluate whether airway responsiveness to methacholine increased after an immediate bronchial response (IBR) and the possible involvement of the beta-adrenoceptor dysfunction in OA-sensitized guinea pigs. Guinea pigs were actively sensitized by aerosolized OA. Following OA exposure, IBR appeared. After IBR when specific airway resistance returned to the base line value, airway responsiveness to methacholine increased significantly. Before OA exposure, propranolol induced bronchoconstriction (PIB) was not provoked, however, after IBR, PIB was provoked and the guinea pigs died because of severe bronchoconstriction. These results suggest that airway responsiveness to methacholine increases significantly after IBR. Furthermore, the dysfunction of the beta-adrenoceptor may be a mechanism of this hyperresponsiveness in OA-sensitized guinea pigs.

How should patients 80 years of age or older with colorectal carcinoma be treated? Long-term and short-term outcome and postoperative cytokine levels.

PURPOSE: The aim of this study was to determine how extensive resection affects operative morbidity, mortality, and long-term survival in elderly patients with colorectal cancer. METHODS: A total of 119 patients 80 years of age or older were given a diagnosis of colorectal carcinoma at our hospital between 1985 and 1997. Eleven patients who did not undergo surgery were excluded. The remaining 108 patients underwent laparotomy and were reviewed. Serum levels of interleukin-6 were measured perioperatively in 22 patients to assess the degree of operative stress. RESULTS: Potentially curative resection was performed in 64 (88.9 percent) of the 72 patients in the active performance status group and 13 (36.1 percent) of the 36 patients in the sedentary performance status group (P < 0.001). The in-hospital mortality rate was 8.3 percent in group the active performance status group and 38 percent in the sedentary performance status group (P = 0.007). Patients in the sedentary performance status group and those who underwent emergency operations had higher levels of IL-6 than patients in the active performance status group or those who underwent elective operations. CONCLUSIONS: Preoperative performance status, operative curability, and tumor stage have a significant impact on outcome in patients with colorectal cancer who are 80 years of age or older. Knowledge of early postoperative response of IL-6 is useful in predicting postoperative mortality and morbidity in this subgroup of patients.

Randomized, controlled trial of lateral node dissection vs. nerve-preserving resection in patients with rectal cancer after preoperative radiotherapy.

PURPOSE: The effectiveness of preoperative radiation therapy for advanced lower rectal carcinoma to preserve the function of pelvic organs and reduce local recurrences was examined in a prospective, randomized, controlled study. METHODS: Fifty-one patients with a diagnosis of localized and resectable adenocarcinoma of the lower rectum undergoing 50 Gy of preoperative radiotherapy were recruited into the trial between April 1993 and March 1995. The patients were randomly allocated to complete autonomic nerve-preserving surgery without lateral node dissection (D1), or surgery with dissection of the lateral lymph nodes including autonomic nerves (D2) followed by oral administration of carmofur for one year. RESULTS: No difference was observed in either survival or disease-free survival between D1 and D2 groups. There was no difference between the two groups in terms of recurrence rate. A significant difference was observed in urinary and sexual function (P = 0.02 and 0.02, respectively) one year after surgery between D1 and D2 groups. CONCLUSION: This study suggests that lateral node dissection is not necessary in terms of curability for patients with advanced carcinoma of the lower rectum who undergo preoperative radiotherapy.

Local immunity and metastasis of colorectal carcinoma.

The subsets of tumor-infiltrating lymphocytes (TIL) and prostaglandin (PG) E2 were measured in the resected tissues of 32 colorectal cancers without metastasis and 14 with metastasis in order to investigate the local immunity in metastasis of colorectal carcinoma. Subsets of TIL (Leu 1, Leu 2a, Leu 3a, Leu 10, Leu 11b, IL-2 receptor) were detected by immunohistochemical staining of frozen tissues. The number of positive cells was counted and expressed as number positive per 250 x 250 microns 2. The numbers of T cells (Leu 1) and natural killer cells (Leu 11b) were larger in early cancers and decreased in parallel with the presence of metastasis (control [n = 9]: 89 +/- 28, 6 +/- 4; early cancers [n = 9]: 269 +/- 112*, 76 +/- 56*; advanced cancers without metastasis [n = 11]: 182 +/- 80*, 56 +/- 59*; advanced cancers with metastasis [n = 11]: 76 +/- 42*, 26 +/- 21; values are mean +/- SD; * P less than 0.05, ANOVA). The level of PG E2 from the draining vein (V) measured by radioimmunoassay was higher than that from the feeding artery (A) (119.1 +/- 14.3 vs. 15.4 +/- 1.9 pg/ml; P less than 0.001). The PG E2 V/A ratio of cancers with metastasis was significantly higher than that of those without metastasis (13.2 +/- 2.4 vs. 5.6 +/- 0.8; P less than 0.001). TIL was decreased in parallel with the increase of PG E2 V/A ratio. We conclude that TIL and PG E2 may play an important role in metastasis of colorectal carcinoma and that PG E2 has an adverse effect in suppressing local immunity and enhancing metastasis.

Limitation of endorectal ultrasonography: what does a low lesion more than 5 mm in size correspond to histologically?

PURPOSE: The accuracy of diagnosis of metastatic lymph nodes for rectal carcinoma has not improved. A low echoic lesion more than 5 mm in size detected by endorectal ultrasonography is preoperatively assessed as a metastatic lymph node. What does a low echoic lesion more than 5 mm in size correspond to histologically? METHODS: Forty-seven patients with lower rectal carcinoma (Term I, 1986-1990) and 48 patients with lower rectal carcinoma (Term II, 1991-1995) undergoing endorectal ultrasonography were reviewed. Surgically resected rectal specimens from 40 patients with rectal carcinoma underwent ultrasonography. Low echoic lesions more than 5 mm in size were taken from resected specimens and were stained with hematoxylin, followed by histologic examination. RESULTS: The accuracy of diagnosis of metastatic lymph nodes of Term II was not higher than that of Term I. Twenty-five low echoic lesions were detected by ultrasonography using 40 resected specimens. Seventeen of these 25 lesions (68 percent) were histologically confirmed to be metastatic lymph nodes. One of 25 (4 percent) was found to be lymphadenitis with no metastasis. Among the 25 lesions, 5 (20 percent) were histologically confirmed to be tumor deposits, 2 (8 percent) fatty tissue, and 22 (88 percent) carcinoma. CONCLUSIONS: Low echoic lesions were histologically confirmed to be tumor deposits (20 percent) and metastatic lymph nodes (68 percent). Although these data show deficiencies and limitations of the accuracy of endorectal ultrasonography, they compare favorably with other series, and no other current technology can show similar accuracy.

Small spot sign of rectal carcinoma by endorectal ultrasonography: histologic relation and clinical impact on postoperative recurrence.

PURPOSE: We observed small spots at the margin of rectal carcinomas on endorectal ultrasonography. Our aim was to study the relationship between ultrasonographic evidence of these spots and histologic characteristics of disease and postoperative recurrence. PATIENTS AND METHODS: The study group comprised 55 patients, 36 men and 19 women, with rectal carcinoma as confirmed by biopsy. The patients were followed up at three-month intervals for six months to three years and six months after the operation. Endorectal ultrasonography was performed in the usual manner. Surgically resected specimens were stained with hematoxylin and eosin and histologically examined. Vessel invasion was graded from 0 (not invasive) to 3 (most invasive). RESULTS: Among the 55 patients studied, 3 had Stage T2,N0,M0 rectal carcinomas and 35 had Stage T3,N0,M0 carcinomas, 5 (14.3 percent) of whom had echographic evidence of small spots. Thirteen patients had Stage T3,N1/2,M0 carcinomas, comprising 12 (92.3 percent) with small spots, and four patients had T3,N1/2,M1 carcinomas, all with small spots. Vessel invasion of Grade 2 or higher was observed around the carcinomas in 20 of 21 patients who had small spots. Ten of 13 patients with many spots at the margin of the carcinoma (a spot grade of ++) histologically had marked venous or lymphatic invasion (an invasion Grade 3). The presence of small spots was closely associated with massive venous or lymphatic invasion (a vessel invasion grade of 2 or more). Four patients had synchronous liver metastases, and small spots were found in all four. Distant metastases and local recurrence were found in 11 of 21 patients with small spots. We found no recurrence in any patient without small spots on endorectal ultrasonography. CONCLUSIONS: Small spots indicate the presence of massive venous or lymphatic invasion and a high risk of postoperative recurrence.

Overexpression of p53 protein and histologic grades of dysplasia in colorectal adenomas.

PURPOSE: To clarify the relation between tumor-suppressor gene p53 expression and histologic grades of dysplasia in colorectal adenomas, we performed immunohistochemical analysis in a series of 59 colorectal polyps and 40 advanced carcinomas. METHODS: Adenomatous polyps were stained by hematoxylin and eosin and classified into mild, moderate, and severe dysplasia (intramucosal carcinoma), according to the World Health Organization's classification. RESULTS: p53 was positive in 7.1 percent (2/28) of mild, 29.4 percent (5/17) of moderate, and 62.5 percent (5/8) of severe dysplasia. In submucosal and advanced carcinomas, positivity rates were 75 percent (3/4) and 47.5 percent (19/40), respectively. Different staining patterns were found, according to grades of dysplasia. In the adenomas with mild or moderate dysplasia, a few focal crypts showed localized p53-positive staining. Adenomas with severe dysplasia had two different staining types. One was a focal staining type as shown in mild or moderate dysplasia; the other was a diffuse staining type, in which glands with mild or moderate dysplasia, surrounding severe dysplasia area, were also stained. Submucosal and advanced carcinomas showed a strong positive staining in cancer cells only. CONCLUSIONS: Overexpression of p53 protein in adenomas with mild or moderate dysplasia and existence of two types of expression in adenomas with severe dysplasia were observed. These facts suggested the possible existence of different pathways in the adenoma to carcinoma progression.