First urinary tract infections in children: the role of the risk factors proposed by the Italian recommendations.

AIM: In 2009, the Italian society for paediatric nephrology suggested the need for cystography, following a first febrile urinary tract infection (UTI), only in children at high risk for dilating vesicoureteral reflux or in the event of a second infection. The aim of this study was to evaluate the adequacy of the risk factors proposed by the Italian guidelines. METHODS: Children aged 2-36 months, managed by 10 Italian hospitals between 2009 and 2013, with a first febrile UTI were retrospectively evaluated. RESULTS: Four hundred and fourteen children were included: 51% female, mean age eight months. Escherichia coli was responsible of 84% UTIs. 269 children (65%) presented at least one risk factor, thus were further investigated: 44% had a reflux. The presence of a pathogen other than E. coli significantly predicted high-grade reflux, both in the univariate (Odd Ratio 2.52, 95% Confidence Interval 1.32-4.81, p < 0.005) and multivariate analysis (OR 2.74, 95% CI: 1.39-5.41, p: 0.003). 26/145 children (18%) with no risk factors experienced a second UTI, which prompted the execution of cystography, showing a dilating reflux in 11. CONCLUSION: Among the risk factors proposed by the Italian guidelines, only the presence of a pathogen other than E. coli significantly predicted reflux. Cystography can be postponed in children with no risk factors.

Bortezomib-induced peripheral neurotoxicity in human multiple myeloma-bearing mice.

The proteasome inhibitor bortezomib is an antineoplastic drug mainly used for the treatment of multiple myeloma (MM). Despite its effectiveness, bortezomib clinical use is often limited by the onset of peripheral neuropathy (BiPN). To better understand the mechanisms of BiPN several rat and mice models have been proposed, but no studies in MM-bearing animals allowing to test the antitumor activity of the selected schedules and the role of MM by itself in peripheral nervous system damage have been reported to date. Here, we carried out a study using immunodeficient C.B-17/Prkdcscid (SCID) mice injected with RPMI8266 human MM cells and treated with bortezomib 1 mg/kg once a week for five weeks. Animals were assessed with neurophysiological, behavioral and pathological methods and tumor volume measurement was performed along the study. At the end of the study BiPN was evident in bortezomib-treated animals, and this neurotoxic effect was evident using a schedule able to effectively prevent tumor growth. However, neurophysiological and pathological evidence of MM induced peripheral nervous system damage was also reported. This model based on MM-bearing animals is more reliable in the reproduction of the clinical setting and it is, therefore, more suitable than the previously reported models of BiPN to study its pathogenesis. Moreover, it represents an optimal model to test the efficacy of neuroprotective agents and at the same time their non-interference with bortezomib antineoplastic activity.

LH, progesterone, and TSH can stimulate aldosterone in vitro: a study on normal adrenal cortex and aldosterone producing adenoma.

Endocrine factors different from ACTH or angiotensin II can stimulate aldosterone secretion and have a role in the pathophysiology of hyperaldosteronism. Aldosterone may increase in luteotropic/progestogenic and in hypothyroid states; LH and, occasionally, TSH receptors have been detected in normal adrenal cortex and aldosterone-producing adenoma. The aim of the study was to compare adrenal contents of LH and TSH receptors between normal cortex and aldosterone-producing adenoma and to evaluate the ability of LH, its product progesterone, and TSH to stimulate aldosterone secretion in vitro from primary adrenocortical cells. Surgical aldosterone-producing adenoma fragments from 19 patients and adrenal cortex fragments from 10 kidney donors were used for Western blotting and cell cultures. LH (n=26), TSH (n=19) and progesterone (n=8) receptor proteins were investigated; LH receptor-mRNA was also tested in 8 samples. Aldosterone responses in vitro to LH, progesterone, and TSH stimulation were assayed. LH and TSH receptors were more expressed in adenoma than normal cortex (p<0.01, p<0.05, respectively); progesterone receptor was observed in 6/8 samples. Aldosterone increased after in vitro stimulation with LH (5/12 adenoma, 1/7 normal cells), progesterone (4/5 adenoma, 5/6 normal cells), and TSH (3/5 adenoma and 3/5 normal cells). LH and TSH receptors were more expressed in aldosterone producing adenoma than normal adrenal cortex. LH, progesterone, and TSH can stimulate aldosterone in vitro. Similar mechanisms could participate in vivo in the aldosterone increase in lutheotropic, progestogenic, or hypothyroid states and may exist in both normal adrenal cortex and adenoma in responsive individuals.

The effect of Photon Activation Therapy on cisplatin pre-treated human tumour cell lines: comparison with conventional X-ray irradiation.

Cisplatin is an antineoplastic drug widely used for the treatment of several solid tumours. However, the side effects related to cisplatin-based anticancer therapy often outweigh the benefits. Therefore, the identification of new anticancer strategies able to offer a better toxicity profile while maintaining the same level of efficacy as platinum-based treatments would be highly desirable. We assessed the efficacy of synchrotron radiation in triggering the Auger effect in human A549 non-small cell lung cancer and IGROV-1 ovarian cancer cells pre-treated with cisplatin. Cisplatin was chosen as the carrier of platinum atoms in the cells because of its alkylating-like activity and the irradiation was done with monochromatic beams above and below the platinum K-shell edge (78.39 keV). On cisplatin-treated cells, at concentrations allowing 80 percent of cell survival with respect to controls, no differences were observed in cell viability when they were irradiated either above or below the K-shell edge of platinum, suggesting that cisplatin toxicity can mask the enhancement of cell death induced by the irradiation. At lower cisplatin concentrations allowing 95-90 percent of cell survival, an enhancement in cellular death with respect to conventional irradiation conditions was clearly observed in all cancer types when cells were irradiated with beams either above or below the platinum K-shell edge. Our results lend additional support to the suggestion that the Photon Activation Therapy in combination with cisplatin treatment should be further explored in relevant in vivo models of glioma and non-glioma cancer models.

Impacts of standard wine-making process on the survival of Lobesia botrana larvae (Lepidoptera: Tortricidae) in infested grape clusters.

To determine the risk winery waste poses for the spread of Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae) in California, we evaluated the survival of larvae in artificially infested grape clusters (Vitis vinifera L.) processed for wine making. The trial consisted of five treatments: whole cluster pressing to 1 bar (100,000 Pa); whole cluster pressing to 2 bars (200,000 Pa); destemming and berry pressing to 1 bar; destemming and berry pressing to 2 bars; and control. Each treatment was replicated with the following five winegrape varieties: Chardonnay, Sauvignon Blanc, Gewürztraminer, Yellow Muscat, and Cabernet Sauvignon. All winery waste was inspected for larval survival. No live larvae were recovered from any of the treatments in all five varieties; therefore, the hypothesis that green winery waste contributes to the spread of L. botrana was rejected.

Prevention of recurrent respiratory tract infections: a literature review of the activity of the bacterial lysate Lantigen B.

OBJECTIVE: Lantigen B, a bacterial lysate, was developed in the 1960s and showed a prophylactic effect in patients with recurrent respiratory tract infections. The objective of this article is to review the literature to update the efficacy and safety profile of Lantigen B in preventing recurrent respiratory tract infections (RRTI). MATERIALS AND METHODS: Articles available from international data banks and producing company archives were used. Only clinical studies providing a control group were considered. The effects of Lantigen B on the number of infectious episodes or comparable parameters were analyzed. RESULTS: 22 randomized clinical trials on 4,571 patients published between 1963 and 2014, with different methodologic accuracy, consistently demonstrated that Lantigen B reduced RRTI vs. placebo (RR -0.47; 95% CI = -0.38 to -0.56). The RR always favored Lantigen B in all the other subsets analyzed in adults with RRTI (RR = -0.48; 95% CI = - 0.33 to -0.62) and children (RR = -0.490; 95% CI = - 0.36 to -0.61). Unfortunately, some studies performed in the past evaluated a small number of patients, and clinical procedures were not always performed according to the more recent good clinical practices. Despite these evident limitations of considered studies, the response frequency has remained almost unchanged since the first articles in the 1960s. CONCLUSIONS: These data confirm the efficacy of Lantigen B alone in the prophylaxis of acute respiratory infections in adults and children but also suggest that Lantigen B, used with novel therapeutic strategies, can further improve clinical outcomes.

Chest wall radiotherapy with volumetric modulated arcs and the potential role of flattening filter free photon beams.

PURPOSE: The goal of the work was to assess the role of RapidArc treatments in chest wall irradiation after mastectomy and determine the potential benefit of flattening filter free beams. METHODS AND MATERIAL: Planning CT scans of 10 women requiring post-mastectomy chest wall radiotherapy were included in the study. A dose of 50 Gy in 2 Gy fractions was prescribed. Organs at risk (OARs) delineated were heart, lungs, contralateral breast, and spinal cord. Dose-volume metrics were defined to quantify the quality of concurrent treatment plans assessing target coverage and sparing of OARs. Plans were designed for conformal 3D therapy (3DCRT) or for RapidArc with double partial arcs (RA). RapidArc plans were optimized for both conventional beams as well as for unflattened beams (RAF). The goal for this planning effort was to cover 100% of the planning target volume (PTV) with ≥ 90% of the prescribed dose and to minimize the volume inside the PTV receiving > 105% of the dose. The mean ipsilateral lung dose was required to be lower than 15 Gy and V(20 Gy) < 22%. Contralateral organ irradiation was required to be kept as low as possible. RESULTS: All techniques met planning objectives for PTV and for lung (3DCRT marginally failed for V(20 Gy)). RA plans showed superiority compared to 3DCRT in the medium to high dose region for the ipsilateral lung. Heart irradiation was minimized by RAF plans with ~4.5 Gy and ~15 Gy reduction in maximum dose compared to RA and 3DCRT, respectively. RAF resulted in superior plans compared to RA with respect to contralateral breast and lung with a reduction of ~1.7 Gy and 1.0 Gy in the respective mean doses. CONCLUSION: RapidArc treatment resulted in acceptable plan quality with superior ipsilateral tissue sparing compared to traditional techniques. Flattening filter free beams, recently made available for clinical use, might provide further healthy tissue sparing, particularly in contralateral organs, suggesting their applicability for large and complex targets.

Small field segments surrounded by large areas only shielded by a multileaf collimator: comparison of experiments and dose calculation.

PURPOSE: Complex radiotherapy fields delivered using a tertiary multileaf collimator (MLC) often feature small open segments surrounded by large areas of the beam only shielded by the MLC. The aim of this study was to test the ability of two modern dose calculation algorithms to accurately calculate the dose in these fields which would be common, for example, in volumetric modulated arc treatment (VMAT) and study the impact of variations in dosimetric leaf gap (DLG), focal spot size, and MLC transmission in the beam models. METHODS: Nine test fields with small fields (0.6-3 cm side length) surrounded by large MLC shielded areas (secondary collimator 12 × 12 cm(2)) were created using a 6 MV beam from a Varian Clinac iX linear accelerator with 120 leaf MLC. Measurements of output factors and profiles were performed using a diamond detector (PTW) and compared to two dose calculations algorithms anisotropic analytical algorithm [(AAA) and Acuros XB] implemented on a commercial radiotherapy treatment planning system (Varian Eclipse 10). RESULTS: Both calculation algorithms predicted output factors within 1% for field sizes larger than 1 × 1 cm(2). For smaller fields AAA tended to underestimate the dose. Profiles were predicted well for all fields except for problems of Acuros XB to model the secondary penumbra between MLC shielded fields and the secondary collimator. A focal spot size of 1 mm or less, DLG 1.4 mm and MLC transmission of 1.4% provided a generally good model for our experimental setup. CONCLUSIONS: AAA and Acuros XB were found to predict the dose under small MLC defined field segments well. While DLG and focal spot affect mostly the penumbra, the choice of correct MLC transmission will be essential to model treatments such as VMAT accurately.

Definition of parameters for quality assurance of flattening filter free (FFF) photon beams in radiation therapy.

PURPOSE: Flattening filter free (FFF) beams generated by medical linear accelerators have recently started to be used in radiotherapy clinical practice. Such beams present fundamental differences with respect to the standard filter flattened (FF) beams, making the generally used dosimetric parameters and definitions not always viable. The present study will propose possible definitions and suggestions for some dosimetric parameters for use in quality assurance of FFF beams generated by medical linacs in radiotherapy. METHODS: The main characteristics of the photon beams have been analyzed using specific data generated by a Varian TrueBeam linac having both FFF and FF beams of 6 and 10 MV energy, respectively. RESULTS: Definitions for dose profile parameters are suggested starting from the renormalization of the FFF with respect to the corresponding FF beam. From this point the flatness concept has been translated into one of "unflatness" and other definitions have been proposed, maintaining a strict parallelism between FFF and FF parameter concepts. CONCLUSIONS: Ideas for quality controls used in establishing a quality assurance program when introducing FFF beams into the clinical environment are given here, keeping them similar to those used for standard FF beams. By following the suggestions in this report, the authors foresee that the introduction of FFF beams into a clinical radiotherapy environment will be as safe and well controlled as standard beam modalities using the existing guidelines.

Salbutamol: how does it enter smooth muscle cells?

Polyspecific organic cation transporters (OCTs) in human cell membranes are involved in the uptake, distribution and excretion of cationic compounds. Although their relevance to drug disposition in the liver, small intestine and kidney has been investigated previously, less is known about the influence of these transporters on the pharmacokinetics and pharmacodynamics of inhaled drugs. Drugs that are commonly administered by inhalation for the treatment of respiratory diseases, such as glucocorticoids and cationic ß(2)-agonists, might interact with several of these transporters, which are strongly expressed on the surfaces of airway epithelial cells. We evaluated the expression of OCT3 and measured the in vitro uptake of the short-acting ß(2)-agonist salbutamol (SALB), alone or in combination with corticosterone (CS) and beclomethasone dipropionate (BDP), by bronchial smooth muscle cells. Our results showed that these cells express the OCT3 transporter and that SALB enters the cell in a transporter-independent fashion. Moreover, CS and BDP have different activities on SALB transport inside the cell. CS increases SALB transport and BDP decreases SALB transport, although neither of these effects are statistically significant. A better understanding of these mechanisms might lead to the improved treatment of airway diseases.

Sr isotope measurements in beef-analytical challenge and first results.

The strontium isotope ratio ((87)Sr/(86)Sr) in beef, derived from 206 European cattle, has been measured. These cattle were located in 12 different European regions within France, Germany, Greece, Ireland, Italy, Spain and the UK. As animal protein is known to be a difficult material on which to conduct Sr isotope analysis, several investigations were undertaken to develop and improve the sample preparation procedure. For example, Sr isotope analysis was performed directly on freeze-dried meat and defatted dry mass from the same samples. It was found that enormous differences-sometimes exceeding the measurement uncertainty-could occur between the fractions and also within one sample even if treated in the same manner. These variations cannot be definitely allocated to one cause but are most likely due to inhomogeneities caused by physiological and biochemical processes in the animals as post mortem contamination during analytical processing could be excluded. For further Sr isotope measurements in meat, careful data handling is recommended, and for the authentic beef samples within this project, it was decided to use only freeze-dried material. It can be demonstrated, however, that Sr isotope measurements in beef proteins are a valuable tool for authentication of geographic origin. Although partly overlapping, some of the European sampling sites could be discriminated even by only using (87)Sr/(86)Sr.

Tubulin binding potentially clears up Bortezomib and Carfilzomib differential neurotoxic effect.

Proteasome inhibitors (PIs) represent the gold standard in the treatment of multiple myeloma. Among PIs, Bortezomib (BTZ) is frequently used as first line therapy, but peripheral neuropathy (PN), occurring approximately in 50% of patients, impairs their life, representing a dose-limiting toxicity. Carfilzomib (CFZ), a second-generation PI, induces a significantly less severe PN. We investigated possible BTZ and CFZ off-targets able to explain their different neurotoxicity profiles. In order to identify the possible PIs off-targets we used the SPILLO-PBSS software that performs a structure-based in silico screening on a proteome-wide scale. Among the top-ranked off-targets of BTZ identified by SPILLO-PBSS we focused on tubulin which, by contrast, did not turn out to be an off-target of CFZ. We tested the hypothesis that the direct interaction between BTZ and microtubules would inhibit the tubulin alfa GTPase activity, thus reducing the microtubule catastrophe and consequently furthering the microtubules polymerization. This hypothesis was validated in a cell-free model, since BTZ (but not CFZ) reduces the concentration of the free phosphate released during GTP hydrolysis. Moreover, NMR binding studies clearly demonstrated that BTZ, unlike CFZ, is able to interact with both tubulin dimers and polymerized form. Our data suggest that different BTZ and CFZ neurotoxicity profiles are independent from their proteasome inhibition, as demonstrated in adult mice dorsal root ganglia primary sensory neurons, and, first, we demonstrate, in a cell free model, that BTZ is able to directly bind and perturb microtubules.

T cell receptor (TCR) structure of autologous melanoma-reactive cytotoxic T lymphocyte (CTL) clones: tumor-infiltrating lymphocytes overexpress in vivo the TCR beta chain sequence used by an HLA-A2-restricted and melanocyte-lineage-specific CTL clone.

HLA-A2+ melanomas express common melanoma-associated antigens (Ags) recognized in vitro by autologous cytotoxic T lymphocytes (CTL). However, it is not known whether tumor Ags can drive in vivo a selective accumulation/expansion of Ag-specific, tumor-infiltrating T lymphocytes (TIL). Therefore, to evaluate this possibility, 39 CTL clones isolated from several independent mixed lymphocyte tumor cultures (MLTC) of TIL and peripheral blood lymphocytes (PBL) of an HLA-A2+ melanoma patient and selected for T cell receptor (TCR)-dependent, HLA-restricted tumor lysis, were used for analysis of TCR alpha and beta chain structure by the cDNA polymerase chain reaction (PCR) technique with variable gene-specific primers followed by sequencing. Despite absence of oligoclonality in fresh TIL and PBL, as well as in T cells of day 28 MLTC (day of cloning), sequence analysis of TCR alpha and beta chains of TIL clones revealed a dominance of a major category of melanoma-specific, HLA-A2-restricted T cells expressing a V alpha 8.2/J alpha AP511/C alpha and V beta 2.1/D beta 1/J beta 1.1/C beta 1 TCR. The same TCR was also found in 2 out of 14 PBL clones. The other PBL clones employed a V alpha 2.1 gene segment associated with either V beta 13.2, 14, or w22. Clones A81 (V alpha 2.1/J alpha IGRJ alpha 04/C alpha and V beta 14/D beta 1/J beta 1.2/C beta 1) and A21 (V alpha 8.2/J alpha AP511/C alpha and V beta 2.1/D beta 1/J beta 1.1/C beta 1), representative of the two most frequent TCR of PBL and TIL, respectively, expressed different lytic patterns, but both were HLA-A2 restricted and lysed only HLA-A2+ melanomas and normal melanocytes, thus indicating recognition of two distinct HLA-A2-associated and tissue-related Ags. Finally, by the inverse PCR technique, the specific TCR beta chain (V beta 2.1/D beta 1/J beta 1.1/C beta 1) expressed by the dominant TIL clone was found to represent 19 and 18.4% of all V beta 2 sequences expressed in the fresh tumor sample and in the purified TIL, respectively, but < 0.19% of V beta 2+ sequences expressed in PBL. These results are consistent with the hypothesis that a clonal expansion/accumulation of a melanocyte-lineage-specific and HLA-A2-restricted T cell clone occurred in vivo at the site of tumor growth.

Translation of a retained intron in tyrosinase-related protein (TRP) 2 mRNA generates a new cytotoxic T lymphocyte (CTL)-defined and shared human melanoma antigen not expressed in normal cells of the melanocytic lineage.

We report here the identification of a new shared human melanoma antigen recognized by a human leukocyte antigen (HLA)-A*68011-restricted cytotoxic T lymphocyte clone (CTL 128). The cDNA encoding this antigen is composed of a partially spliced form of the melanocyte differentiation antigen tyrosinase-related protein (TRP)-2, containing exons 1-4 with retention of intron 2 and part of intron 4 (TRP-2-INT2). The sequence coding for the antigenic epitope is located at the 5' end of intron 2 and is available for translation in the same open reading frame of the fully spliced TRP-2 mRNA. This peptide is also recognized by CTL 128 when presented by the HLA-A*3301, a member of the HLA-A3-like supertype that includes the HLA-A*68011. Quantitative reverse transcription PCR analysis carried out on total and/or cytoplasmic mRNA demonstrated that, in contrast to the fully spliced TRP-2 mRNA expressed in melanomas, normal skin melanocytes, and retina, the TRP-2-INT2 mRNA could be detected at significant levels in melanomas but not in normal cells of the melanocytic lineage. Instead, in these normal samples, both the spliced and the unspliced transcript of gp100 were expressed at high levels. Absence of endogenous TRP-2-INT2 expression in melanocytes was also confirmed by lack of recognition of HLA-A*68011-transduced, TRP-2(+) melanocyte lines by CTL 128. These results indicate that a partially spliced form of a differentiation antigen mRNA, present in the cytoplasmic compartment of neoplastic but not normal cells of the melanocytic lineage, can be the source of a melanoma-restricted T cell epitope.

Effects of extra-fine inhaled beclomethasone/formoterol on both large and small airways in asthma.

BACKGROUND: Airway inflammation in asthma involves both large and small airways, and the combination of inhaled corticosteroids (ICS) and long acting beta-2 agonists (LABA) is the mainstay of therapy. Available inhaled combinations differ in terms of drug delivery to the lung and the ability to reach small airways. AIM: To evaluate whether treatment with an extra-fine inhaled combination provides additional effects vs a nonextra-fine combination on airway function. METHODS: After a 1- to 4-week run-in period, patients with asthma were randomized to a double blind, double dummy, 12-week treatment with either extra-fine beclomethasone/formoterol (BDP/F) 400/24 microg daily or fluticasone propionate/salmeterol (FP/S) 500/100 microg daily. Methacholine (Mch) bronchoprovocation challenge and single breath nitrogen (sbN2) test were performed. RESULTS: Thirty patients with asthma (15 men), mean age 43, mean forced expiratory volume in the first second (FEV(1)) 71.4% of predicted, were included. A significant increase (P < 0.01) versus baseline was observed in predose FEV(1) in both BDP/F and FP/S groups (0.37 +/- 0.13 l and 0.36 +/- 0.12 l, respectively). PD(20)FEV(1) Mch improved significantly from 90.42 (+/-30.08) microg to 432.41 (+/-122.71) microg in the BDP/F group (P = 0.01) but not in the FP/S group. A trend toward improvement vs baseline was observed for BDP/F in closing capacity (CC), whereas no differences were recorded in other sbN(2) test parameters. CONCLUSION: The findings of this pilot study suggest that an extra-fine inhaled combination for the treatment of asthma has beneficial effects on both large and small airways function as expressed by Mch and sbN(2) tests.

The Swiss IMRT dosimetry intercomparison using a thorax phantom.

PURPOSE: In 2008, a national intensity modulated radiation therapy (IMRT) dosimetry intercomparison was carried out for all 23 radiation oncology institutions in Switzerland. It was the aim to check the treatment chain focused on the planning, dose calculation, and irradiation process. METHODS: A thorax phantom with inhomogeneities was used, in which thermoluminescence dosimeter (TLD) and ionization chamber measurements were performed. Additionally, absolute dosimetry of the applied beams has been checked. Altogether, 30 plan-measurement combinations have been used in the comparison study. The results have been grouped according to dose calculation algorithms, classified as "type a" or "type b," as proposed by Kntis et al. ["Comparison of dose calculation algorithms for treatment planning in external photon beam therapy for clinical situations," Phys. Med. Biol. 51, 5785-5807 (2006)]. RESULTS: Absolute dosimetry check under standard conditions: The mean ratio between the dose derived from the single field measurement and the stated dose, calculated with the treatment planning system, was 1.007 +/- 0.010 for the ionization chamber and 1.002 +/- 0.014 (mean+/- standard deviation) for the TLD measurements. IMRT Plan Check: In the lung tissue of the planning target volume, a significantly better agreement between measurements (TLD, ionization chamber) and calculations is shown for type b algorithms than for type a (p <0.001). In regions outside the lungs, the absolute differences between TLD measured and stated dose values, relative to the prescribed dose, [(Dm-Ds)/Dprescribed], are 1.9 +/- 0.4% and 1.4 +/- 0.3%, respectively. These data show the same degree of accuracy between the two algorithm types if low-density medium is not present. CONCLUSIONS: The results demonstrate that the performed intercomparison is feasible and confirm the calculation accuracies of type a and type b algorithms in a water equivalent and low-density environment. It is now planned to offer the intercomparison on a regular basis to all Swiss institutions using IMRT techniques.

Bronchial vascular remodelling in patients with COPD and its relationship with inhaled steroid treatment.

BACKGROUND: Only a few studies have evaluated microvascular changes and proangiogenetic mediators in the bronchial mucosa of patients with chronic obstructive pulmonary disease (COPD), and the results have been discordant. Furthermore, the role of inhaled corticosteroids (ICS) in COPD has not been extensively studied. A study was undertaken to evaluate vascular remodelling, its relationship with inflammatory cells and treatment effects in the bronchial mucosa of patients with COPD. METHODS: The study comprised three groups: (1) 10 non-treated patients with COPD (COPD); (2) 10 patients with COPD treated with nebulised beclomethasone dipropionate 1600-2400 mug daily (equivalent to 800-1200 mug via metered dose inhaler) (COPD/ICS); and (3) 8 control subjects (CS). Bronchial biopsies were evaluated for number and size of vessels and vascular area. Specimens were also examined for vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and transforming growth factor beta (TGF-beta) expression and inflammatory cell counts were performed. RESULTS: Vascular area, vessel size, VEGF+ cells, bFGF+ cells and TGF-beta+ cells were significantly increased in the COPD group compared with the COPD/ICS and CS groups (all p<0.05). In addition, bFGF+ cells were significantly increased in the COPD/ICS group compared with the CS group, and CD8+ and CD68+ cells were significantly increased in the COPD group compared with the COPD/ICS and CS groups (p<0.05). In the COPD group the VEGF+ cells correlated with the number of vessels (p<0.05), vascular area (p<0.01) and vessel size (p<0.05), and TGF-beta+ cells correlated significantly with vascular area (p<0.05). CONCLUSION: Bronchial vascular remodelling in patients with COPD is mainly related to morphological changes of the mucosal microvessels rather than to new vessel formation, and may be reduced in patients treated with steroids.

Regular vs prn nebulized treatment in wheeze preschool children.

BACKGROUND: International guidelines recommend regular treatment with inhaled glucocorticoids for children with frequent wheezing; however, prn inhaled bronchodilator alone or in combination with glucocorticoid is also often used in practice. We aimed to evaluate whether regular nebulized glucocorticoid plus a prn bronchodilator or a prn nebulized bronchodilator/glucocorticoid combination is more effective than prn bronchodilator alone in preschool children with frequent wheeze. METHODS: Double-blind, double-dummy, randomized, parallel-group trial. After a 2-week run-in period, 276 symptomatic children with frequent wheeze, aged 1-4 years, were randomly assigned to three groups for a 3-month nebulized treatment: (1) 400 microg beclomethasone bid plus 2500 microg salbutamol prn; (2) placebo bid plus 800 microg beclomethasone/1600 microg salbutamol combination prn; (3) placebo bid plus 2500 microg salbutamol prn. The percentage of symptom-free days was the primary outcome measure. Secondary outcomes included symptom scores, use of relief medication and exacerbation frequency. RESULTS: As compared with prn salbutamol (61.0 +/- 24.83 [SD]), the percentage of symptom-free days was higher with regular beclomethasone (69.6%, SD 20.89; P = 0.034) but not with prn combination (64.9%, SD 24.74). Results were no different in children with or without risk factors for developing persistent asthma. The effect of prn combination was no different from that of regular beclomethasone on the primary and on several important secondary outcomes. CONCLUSIONS: Regular inhaled glucocorticoid is the most effective treatment for frequent wheezing in preschool children. However, prn bronchodilator/glucocorticoid combination might be an alternative option, but it requires further study.

Anti-proliferative and anti-remodelling effect of beclomethasone dipropionate, formoterol and salbutamol alone or in combination in primary human bronchial fibroblasts.

BACKGROUND: Bronchial asthma is characterized by lower airway inflammation and remodelling. Anti-inflammatory treatment with inhaled corticosteroids (ICS) provides the mainstay of asthma therapy together with bronchodilation induced by short- and long-acting inhaled beta(2)-agonists. Lower airway fibroblasts may play a critical role in airway inflammation and remodelling, suggesting that they might represent an important target for the major anti-asthmatic drugs. The aim of our study was to investigate the effects of beclomethasone dipropionate (BDP), salbutamol and formoterol either alone or in combination on in vitro cultures of human bronchial fibroblasts. METHODS: Fibroblasts were cultured in the presence of pro-inflammatory and proliferative stimuli, BDP, salbutamol and formoterol. The effects of drugs on cell proliferation were ascertained by (3)H-thymidine incorporation. CD90 and CD44 expression were detected by flow cytometry and fibronectin secretion using the enzyme-linked immunosorbent assay technique. RESULTS: This study showed that BDP alone has significant anti-proliferative effects on lung fibroblasts treated with basic fibroblast growth factor and the combination of BDP with formoterol or salbutamol strengthen these effects. Short-acting beta(2)-agonist (SABA) or long-acting beta(2)-agonist (LABA) by themselves did not show any significant effect on the different cultures. CD44 and CD90 expression and fibronectin production were modulated by pro-inflammatory and proliferative stimuli; the addition of the drugs brought them back near to the basal level. CONCLUSIONS: From this in vitro study, we can conclude that BDP, when combined with salbutamol or formoterol, exhibits enhanced anti-remodelling activity in bronchial fibroblasts, providing new insights on the additive effects of ICS and SABAs and LABAs for asthma therapy.

Glycosylated simple phenolic profiling of food tannins using high resolution mass spectrometry (Q-Orbitrap).

Tannins are polyphenolic compounds extensively present in plants and used by food industry as processing aids. Due to the heterogeneity of plant sources, actions involved in food processing and tannin commercial costs can be different. In the last years different approaches aimed at correctly identifying the tannin botanical origin have been developed, in order to satisfy the industry's request to verify product labels. This work aimed to define the glycosidic simple phenolic profile of a large selection of monovarietal commercial tannins of different origin, using a high-resolution untargeted approach. Using accurate mass, isotopic pattern and MS/MS fragmentation, 167 precursors, 89 as monoglycosylated and 78 as diglycosylated derivatives were tentatively identified in tannins, validating the untargeted approach with 3 custom-synthesized glycosidic precursors. Almost all tannin botanical varieties were shown to be characterised by a specific glycosylated phenolic profile, providing possible tools for tannin classification in the case of glycosylphenol standard availability.