Using a guinea‐pig model of allergic asthma, we investigated the role of nitric oxide (NO) in allergen‐induced airway hyperreactivity after the early asthmatic reaction, by examining the effects of the NO‐synthase inhibitor Nω‐nitro‐L‐arginine methyl ester (L‐NAME) on the responsiveness to methacholine and histamine of isolated perfused tracheae from unchallenged (control) animals and from animals 6 h after ovalbumin challenge. All animals developed airway hyperreactivity to inhaled histamine at 6 h after ovalbumin challenge, with a mean 3.11 ± 0.45 fold increase in sensitivity to the agonist (P < 0.001). In perfused tracheal preparations from the ovalbumin‐challenged guinea‐pigs, the maximal responses (Emax) to methacholine and histamine were significantly enhanced compared to controls, both after intraluminal (IL) and extraluminal (EL) administration of the contractile agonists. In addition, a small but significant increase in the pD2 (‐log10 EC50) for IL and EL methacholine and for IL histamine was observed. As a consequence, the ΔpD2 (EL‐IL) for histamine was slightly decreased from 1.67 ± 0.13 to 1.23 ± 0.14 (P < 0.05). However, the ΔpD2 for methacholine was unchanged (1.85 ± 0.11 and 1.77 ± 0.12, respectively; NS). Incubation of control tracheae with 100 μm L‐NAME (IL) significantly enhanced the Emax for both IL and EL methacholine and histamine to approximately the same degree as observed after ovalbumin challenge, with no effect on the pD2 and ΔpD2 for both agonists. On the contrary, L‐NAME had no effect on Emax and pD2 values of tracheal preparations from ovalbumin‐challenged guinea‐pigs. L‐NAME (10 μm‐l mM) had no effect on methacholine‐induced contraction of isolated tracheal strip preparations obtained from control animals, indicating that L‐NAME has no antimuscarinic effect on tracheal smooth muscle. Histological examination of the intact tracheal preparations indicated epithelial and subepithelial infiltration of eosinophils after ovalbumin challenge. However, no apparent damage of the airway epithelium was observed in these preparations. The results indicate that a deficiency of NO contributes to allergen‐induced airway hyperreactivity after the early asthmatic reaction and that this deficiency appears not to be due to epithelial shedding.

Background-Nitric oxide (NO) is involved in inflammation and host defence of the lung. It has been found in increased concentrations in the airways in asthmatic subjects but its levels in patients with chronic obstructive pulmonary disease (COPD) have not been investigated. A study was undertaken to determine whether markers of NO metabolism (NO in exhaled air, iNOS expression in sputum cells, and nitrite + nitrate (NO 2 -/NO 3 -) in sputum supernatant) are increased in subjects with COPD, and whether they correlate with inflammatory indices in induced sputum. The associations of these markers with smoking were also assessed. Methods-Sixteen subjects with COPD (median age 66 years, median forced expiratory volume in one second (FEV 1 ) 63% predicted, eight current smokers) and 16 healthy subjects (median age 63 years, median FEV 1 113% predicted, eight current smokers) participated in the study. NO was measured during tidal breathing and sputum was induced by inhalation of hypertonic saline. Results-No

SUMMARYTen commercially available rabbit polyclonal anti-NOS antibodies were tested for their immunohistological applicability in normal human, guinea pig, rat, and mouse organs. Most antibodies reacted as expected and described in the literature with various tissues of the investigated species. Several antibodies did not react with the expected cell populations in a certain species, or reacted in previously unknown patterns. In addition, different antibodies to the same isoform rarely detected identical cell populations, even within one species. Most of these unexpected immunoreactivities were observed in bronchial epithelial, glomerular epithelial, and vascular smooth muscle cells. These unexpected results usually occurred when the antibodies were tested in other organs or species than that to which they were originally raised. We therefore strongly recommend the use of anti-NOS antibodies only after careful immunohistological and biochemical analysis of their reactivity in the organ and species to be studied.

BackgroundCapecitabine monotherapy is a treatment option for selected patients with metastatic colorectal cancer (mCRC) and is administered to up to 17% of patients. Data are limited with regard to adverse events and dosing practices associated with capecitabine monotherapy in real-world situations.ObjectivesThe aim of this study was to provide real-world data on adverse event rates and dose adjustments/discontinuations associated with capecitabine monotherapy in patients with mCRC.MethodsThis retrospective study analyzed data from CRC patients scheduled to receive up to eight planned cycles of capecitabine monotherapy between 2009 and 2013 at a single large community hospital in The Netherlands. Data on adverse events (hand-foot syndrome [HFS], gastrointestinal (GI) events, hematological adverse events, and cardiotoxicity), as well as relative dose intensities (RDIs), dose reductions, and discontinuations, were evaluated.ResultsData from 86 patients (45 females; mean age at the start of treatment, 69 years) were included. A total of 46.5% of patients experienced HFS and 44.2% experienced a GI event at some time during treatment. Hematological events and cardiotoxicity were rare. Most patients (77%) started at below the recommended dose, and patients at the lowest dose also had the lowest median RDIs. Dose reductions and discontinuations occurred in 15–25% of patients who experienced HFS or GI event over the course of eight cycles.ConclusionsHFS and GI events were very common in patients treated with capecitabine monotherapy in a real-world clinical setting. Most patients started treatment at below the recommended dose, and 15–25% of patients who had HFS or a GI event had a dose reduction or discontinuation.

SUMMARYWe examined immunopathological changes of podocytes in vivo which, based on in vitro studies, are thought to be relevant for the pathogenesis of renal diseases. We investigated the alterations of podocytes in local inflammation in a recently developed model of pauci-immune necrotizing crescentic glomerulonephritis (NCGN) in the rat. Frozen and plastic embedded kidney sections at different time points of the disease were incubated with antibodies directed to MHC class I, MHC class II, ICAM-1 and to relevant cytokines. Strong glomerular expression of MHC class I, II and ICAM-1 was found within 4 days, and plastic embedded sections clearly demonstrated increased cell membrane staining of podocytes. Increased glomerular interferon-gamma (IFN--y) was detected within 24 h of induction of NCGN, and IL-1,3 and tumour necrosis factor-alpha (TNF-a) were found from day 4. The potency of these cytokines to induce adhesion molecules on podocytes was investigated on rat glomerular epithelial cells in vitro. By using FACS analysis and electron microscopical techniques, we found that the in vivo expression of MHC class I, II and ICAM-l by podocytes could in vitro be simulated by IFN--y. IFN-a weakly induced MHC class I, while IL-lf and TNF-a were ineffective. We hypothesize that podocytes in this in vivo model are important to maintain the local inflammatory process in the glomerulus by expression of relevant adhesion molecules and MHC molecules upon stimulation with specific cytokines. Keywords glomerular visceral epithelial cells MHC intercellular adhesion molecule-l interferon-alpha interferon