Serum endostatin levels are elevated in colorectal cancer and correlate with invasion and systemic inflammatory markers

Background:

Endostatin, a fragment of collagen XVIII, is an endogenous angiogenesis inhibitor with anti-tumour functions. However, elevated circulating endostatin concentrations have been found in several human cancers including colorectal cancer (CRC).

Methods:

Serum endostatin levels were measured by enzyme-linked immunoassay from a series of 143 patients with CRC and from 84 controls, and correlated with detailed clinicopathological features of CRC, serum leukocyte differential count and C-reactive protein (CRP) levels.

Results:

Patients with CRC had higher serum endostatin levels than the controls (P=0.005), and high levels associated with age, tumour invasion through the muscularis propria and poor differentiation, but not with metastases. Endostatin levels showed a positive correlation with the markers of systemic inflammatory response and a negative correlation with the densities of tumour-infiltrating mast cells and dendritic cells. Collagen XVIII was expressed in tumour stroma most strikingly in blood vessels and capillaries, and in the muscle layer of the bowel wall.

Conclusions:

Elevated endostatin levels in CRC correlate with systemic inflammation and invasion through the muscularis propria. Increased endostatin level may be a result of invasion-related cleavage of collagen XVIII expressed in the bowel wall. The negative correlations between serum endostatin and intratumoural mast cells and immature dendritic cells may reflect angiogenesis inhibition by endostatin.     

An improved image analysis method for cell counting lends credibility to the prognostic significance of T cells in colorectal cancer

Numerous immunohistochemically detectable proteins, such as immune cell surface (CD) proteins, vascular endothelial growth factor, and matrix metalloproteinases, have been proposed as potential prognostic markers in colorectal cancer (CRC) and other malignancies. However, the lack of reproducibility has been a major problem in validating the clinical use of such markers, and this has been attributed to insufficiently robust methods used in immunohistochemical staining or its assessment. In this study, we assessed how computer-assisted image analysis might contribute to the reliable assessment of positive area percentage and immune cell density in CRC specimens, and subsequently, we applied the computer-assisted cell counting method in assessing the prognostic value of T cell infiltration in CRC. The computer-assisted analysis methods were based on separating hematoxylin and diaminobenzidine color layers and then applying a brightness threshold using open source image analysis software ImageJ. We found that computer-based analysis results in a more reproducible assessment of the immune positive area percentage than visual semiquantitative estimation. Computer-assisted immune cell counting was rapid to perform and accurate (Pearson r > 0.96 with exact manual cell counts). Moreover, the computer-assisted determination of peritumoral and stromal T cell density had independent prognostic value. Our results suggest that computer-assisted image analysis, utilizing freely available image analysis software, provides a valuable alternative to semiquantitative assessment of immunohistochemical results in cancer research, as well as in clinical practice. The advantages of using computer-assisted analysis include objectivity, accuracy, reproducibility, and time efficiency. This study supports the prognostic value of assessing T cell infiltration in CRC.     

Asthma programme in Finland: comparison of adult asthma referral letters in 1994 and 2001

OBJECTIVES: The aim of the study was to evaluate the quality of asthma-related referral letters at the launch of the Asthma Programme in 1994 and 7 years later in 2001. METHODS: All referrals during 1 year (n=1121 in 1994, n=1136 in 2001) to one pulmonary department were screened in 2001. By the same inclusion criteria of asthma or suspicion of asthma, 624 letters (56% of all) from the year 1994 and 452 (40% of all) from the year 2001 were selected. The quality of study letters was assessed against the previously developed asthma referral letter criteria. Comparison of the referral letter quality in 1994 and 2001 was made. RESULTS: The proportion of poor letters decreased from 63% in 1994 to 44% in 2001, while that of good letters increased from 7% to 22%. Graphics of peak flow follow-up measurements (14% vs. 40%) and spirometry with bronchodilatation test (5% vs. 32%) were included significantly more often as an attachment. CONCLUSION: Lung functions are being measured more often in primary care, indication a more active detection of asthma. The number of asthma-related referrals in relation to all pulmonary consultation referrals decreased and their quality improved during the years of the Asthma Programme.     

CARDIOPROTECTIVE EFFECT OF l-GLUTAMATE IN OBESE TYPE 2 DIABETIC ZUCKER FATTY RATS

• 1 Because diabetic hearts have an increased threshold for cardioprotection by ischaemic preconditioning (IPC), we hypothesized that protection by l ‐glutamate during reperfusion is restricted in Type 2 diabetic hearts. Previously, we found that l ‐glutamate‐mediated postischaemic cardioprotection mimics IPC.
• 2 Rat hearts were studied in a Langendorff preparation perfused with Krebs’–Henseleit solution and subjected to 40 min global no‐flow ischaemia, followed by 120 min reperfusion. l ‐Glutamate (0, 15 and 30 mmol/L) was added to the perfusate during reperfusion of hearts from non‐diabetic (Wistar‐Kyoto) and diabetic (Zucker diabetic fatty (ZDF)) rats, studied at 16 weeks of age. The infarct size (IS)/area‐at‐risk (AAR) ratio was the primary end‐point. Expression of l ‐glutamate excitatory amino acid transporter (EAAT) 1 (mitochondrial) and EAAT3 (sarcolemmal) was determined by quantitative polymerase chain reaction and immunoblotting.
• 3 The ISS/AAR ratio did not differ between control hearts from Wistar‐Kyoto and ZDF rats (0.52 ± 0.03 and 0.51 ± 0.04, respectively; P = 0.90). l ‐Glutamate (15 mmol/L) significantly reduced the IS/AAR ratio in non‐diabetic hearts, but not in diabetic hearts, compared with their respective controls. The higher concentration of l ‐glutamate (30 mmol/L) reduced infarct size in diabetic hearts to the same degree as in non‐diabetic hearts (IS/AAR 0.35 ± 0.03 (P = 0.002) and 0.34 ± 0.03 (P = 0.004), respectively). The mitochondrial l ‐glutamate transporter EAAT1 was downregulated in hearts from ZDF rats at both the mRNA and protein levels (P < 0.0005 and P < 0.0001, respectively). However, there was no change in EAAT3 expression at the protein level. Myocardial l ‐glutamate content was increased by 43% in diabetic hearts (P < 0.0001).
• 4 Hearts from obese diabetic rats have an elevated threshold for metabolic postischaemic cardioprotection by l ‐glutamate. These findings may reflect underlying mechanisms of inherent resistance against additional cardioprotection in the diabetic heart.

     

Frequent mutations of KRAS in addition to BRAF in colorectal serrated adenocarcinoma

Stefanius K, Ylitalo L, Tuomisto A, Kuivila R, Kantola T, Sirniö P, Karttunen T J & Mäkinen M J
(2011) Histopathology 58 , 679–692
Frequent mutations of KRAS in addition to BRAF in colorectal serrated adenocarcinoma Aims: To define the occurrence of KRAS and BRAF mutations, microsatellite instability (MSI), and MGMT and hMLH1 methylation and expression in colorectal serrated adenocarcinoma. Methods and results: KRAS codon 12/13 and 59/61 and BRAF V600E mutations, MSI, and MGMT and hMLH1 methylation and expression in 42 serrated adenocarcinomas and 17 serrated adenomas were compared with those in 59 non‐serrated colorectal carcinomas (CRCs) and nine adenomas. KRAS and BRAF mutations were observed in 45% and 33% of serrated adenocarcinomas and in 27% and 0% of non‐serrated CRCs (P < 0.001). The KRAS c12G→A transition was the predominant type of mutation in serrated adenocarcinomas. Forty‐two per cent of BRAF‐mutated serrated adenocarcinomas showed high‐level MSI (MSI‐H) (P = 0.075), 100% showed hMLH1 methylation (P = 0.001) and 90.9% showed MGMT methylation (P = 0.019). Fifty‐six per cent of serrated adenocarcinomas with microsatellite stability/low‐level microsatellite instability harboured KRAS mutations. In non‐serrated cancers, KRAS mutations were not associated with MSI status. Conclusions: A high combined mutation rate (79–82%) of KRAS and BRAF in serrated adenomas and adenocarcinomas indicates that mitogen‐activated protein kinase activation is a crucial part of the serrated pathway. BRAF mutations are specific for serrated adenocarcinoma and identify a subset of serrated adenocarcinomas with gene methylation and a tendency for MSI‐H. A high frequency of KRAS mutations in serrated adenocarcinomas suggests that a significant proportion of KRAS‐mutated CRCs originate from serrated precursors, thus challenging the traditional model of Vogelstein.     

Predicting arterial stiffness with ambulatory blood pressure: an 11-year follow-up

No prospective data have been published on whether ambulatory blood pressure (BP) works better than casual measurements in predicting arterial stiffness. This study with 11-year follow-up was launched to evaluate the usefulness of ambulatory intra-arterial BP in predicting pulse wave velocity (PWV). Ninety-seven previously healthy men were recruited from a routine physical check-up at baseline. BP was measured with standard cuff and intra-arterial ambulatory methods. Sixty-seven subjects with no antihypertensive medication were enrolled for a visit after a follow-up of 11 years. Arterial stiffness was estimated with PWV derived with impedance cardiography. Ambulatory 24-h systolic blood pressure (SBP) (r = 0.30, P = 0.01), 24-h mean arterial pressure (r = 0.27, P = 0.03), 24-h pulse pressure (r = 0.27, P = 0.03) and daytime SBP (r = 0.26, P = 0.03) were the best BP variables in predicting future PWV. Casual BP values did not bear significant correlations with future PWV. In hierarchical regression analysis, the best predictive value for future PWV was achieved with the model including ambulatory 24-h SBP, smoking (number of cigarettes) and age (adjusted R(2) = 0.26). In conclusion, to our knowledge, this is the only prospective follow-up study to show that ambulatory BP is superior to casual BP measurement in predicting future PWV.     

注射用右兰索拉唑治疗急性胃和/或十二指肠溃疡引起的上消化道出血的有效性及安全性

目的：以注射用兰索拉唑为对照,评价注射用右兰索拉唑15 mg q12 h治疗急性胃和/或十二指肠溃疡引起的上消化道出血的有效性及安全性。方法：选取全国31家研究中心的急性胃和/或十二指肠溃疡引起的上消化道出血患者共202例,按照1∶1随机分配至试验组（注射用右兰索拉唑组）和对照组（注射用兰索拉唑组）。主要疗效终点为72 h有效止血率。对主要疗效终点采用非劣效评价,非劣效性界值δ是10%。结果：有效性方面,全分析数据集分析结果显示：用药72 h后,注射用右兰索拉唑组有效止血率为96.08%(98/102);注射用兰索拉唑组有效止血率为98.00%(98/100),两组率差为-1.92%(95%CI-6.58,2.74)。两组72 h有效止血率差异无统计学意义（P=0.682 9）。两组率差的双侧界值均低于δ(10%),注射用右兰索拉唑非劣于注射用兰索拉唑。安全性方面,试验组的不良事件及不良反应发生率与对照组差异无统计学意义,无非预期不良反应和严重不良反应。主要的不良反应为白细胞计数降低、中性粒细胞计数降低等。结论：注射用右兰索拉唑15 mg q12 h在治疗急性胃和/或十二指肠溃疡引起的...