Up-regulation of autophagy is a mechanism of resistance to chemotherapy and can be inhibited by pantoprazole to increase drug sensitivity.

BACKGROUND: Autophagy is a survival mechanism that allows recycling of cellular breakdown products, particularly in stressed cells. Here we evaluate the hypotheses that up-regulation of autophagy is a common mechanism of resistance to chemotherapy, and that drug resistance can be reversed by inhibiting autophagy with a proton pump inhibitor. METHODS: We exposed human PC3, LNCaP and MCF7 cells to seven clinically-used chemotherapy drugs ± pantoprazole, examined the up-regulation of autophagy and the effect on cellular proliferation by Western Blots, MTS assay and colony-forming assay. The distribution of drug effects and of autophagy was quantified in LNCaP tumor sections in relation to blood vessels and hypoxia by immunohistochemistry using γH2AX, cleaved caspase-3 and p62. RESULTS: All anticancer drugs led to up-regulation of autophagy in cultured tumor cells. Pantoprazole inhibited the induction of autophagy in a time- and dose-dependent manner, and sensitized cancer cells to the seven anti-cancer drugs. Treatment of LNCaP xenografts with paclitaxel induced both DNA damage and autophagy; autophagy was inhibited and markers of toxicity were increased by pantoprazole. CONCLUSIONS: Induction of autophagy is a general mechanism associated with resistance to anticancer drugs and that its inhibition is a promising therapeutic strategy to enhance the effects of chemotherapy and improve clinical outcomes.

Lymphangiogenesis in abdominal aortic aneurysm.

BACKGROUND: Ongoing angiogenesis is implicated in the inflammatory environment that characterizes abdominal aortic aneurysm (AAA). Although lymphangiogenesis has been associated with chronic inflammatory conditions, it has yet to be demonstrated in AAA. The aim was to determine the presence of lymphangiogenesis and to delineate the relationship between inflammation and neovascularization in AAA tissue. METHODS: AAA samples and preoperative computed tomography images were obtained from patients undergoing elective AAA repair. Control samples were age-matched abdominal aortic tissue. Specific immunostains for blood vessels (CD31, CD105), lymphatic vessels (D2-40), vascular endothelial growth factor (VEGF) A and VEGF receptor (VEGFR) 3 allowed characterization and quantitation of vasculature. RESULTS: The AAA wall contained high levels of inflammatory infiltrate; microvascular densities of blood (P < 0·001) and lymphatic (P = 0·003) vessels were significantly increased in AAA samples compared with controls. Maximal AAA vascularity was observed in inflammatory areas, with vessels that stained positively for CD31 (ρ = 0·625, P = 0·017), CD105 (ρ = 0·692, P = 0·009) and D2-40 (ρ = 0·675, P = 0·008) correlating positively with the extent of inflammation. Increased VEGFR-3 and VEGF-A expression was also evident within inflammatory AAA areas. CONCLUSION: These findings demonstrated lymphatic vessel involvement in end-stage AAA disease, which was associated with the degree of inflammation, and confirmed the involvement of neovascularization.

Fine-needle aspiration biopsy versus core-needle biopsy in diagnosing lung cancer: a systematic review.

BACKGROUND: Lung cancer leads cancer-related mortality in the world. The objective of the present systematic review was to compare fine-needle aspiration biopsy (fnab) with core-needle biopsy (cnb) for diagnostic characteristics and yields for diagnosing lung cancer in patients with lung lesions. METHODS: The medline and embase databases (from January 1, 1990, to September 14, 2009), the Cochrane Library (to Issue 4, 2009), and selected guideline Web sites were searched for relevant articles. RESULTS: For overall diagnostic characteristics (benign vs. malignant) of fnab and cnb, the ranges of sensitivity were 81.3%-90.8% and 85.7-97.4% respectively; of specificity, 75.4%-100.0% and 88.6%-100.0%; and of accuracy, 79.7%-91.8% and 89.0%-96.9%. For specific diagnostic characteristics of fnab and cnb (identifying the histologic subtype of malignancies or the specific benign diagnoses), the ranges of sensitivity were 56.3%-86.5% and 56.5-88.7% respectively; of specificity, 6.7%-57.1% and 52.4%-100.0%; and of accuracy, 40.4%-81.2% and 66.7%-93.2%. Compared with fnab, cnb did not result in a higher complication rate (pneumothorax or hemoptysis). No study has yet compared the diagnostic yields of fnab and of cnb for molecular predictive-marker studies in patients with lung lesions. DISCUSSION AND CONCLUSIONS: The evidence is currently insufficient to support a difference between fnab and cnb in identifying lung malignancies in patients with lung lesions. Compared with fnab, cnb might have a higher specificity to diagnose specific benign lesions. Well-designed, good-quality studies comparing fnab with cnb for diagnostic characteristics and yields in diagnosing lung cancer should be encouraged.

Efficacy assessment of sustained intraperitoneal paclitaxel therapy in a murine model of ovarian cancer using bioluminescent imaging.

We evaluated the pre-clinical efficacy of a novel intraperitoneal (i.p.) sustained-release paclitaxel formulation (PTX(ePC)) using bioluminescent imaging (BLI) in the treatment of ovarian cancer. Human ovarian carcinoma cells stably expressing the firefly luciferase gene (SKOV3(Luc)) were injected i.p. into SCID mice. Tumour growth was evaluated during sustained or intermittent courses of i.p. treatment with paclitaxel (PTX). In vitro bioluminescence strongly correlated with cell survival and cytotoxicity. Bioluminescent imaging detected tumours before their macroscopic appearance and strongly correlated with tumour weight and survival. As compared with intermittent therapy with Taxol, sustained PTX(ePC) therapy resulted in significant reduction of tumour proliferation, weight and BLI signal intensity, enhanced apoptosis and increased survival times. Our results demonstrate that BLI is a useful tool in the pre-clinical evaluation of therapeutic interventions for ovarian cancer. Moreover, these results provide evidence of enhanced therapeutic efficacy with the sustained PTX(ePC) implant system, which could potentially translate into successful clinical outcomes.

Effect of obesity on airway inflammation: a cross-sectional analysis of body mass index and sputum cell counts.

BACKGROUND: Several observational studies have demonstrated an association between obesity and asthma. Studies evaluating exhaled nitric oxide levels and obesity have revealed that a higher body mass index (BMI) is associated with elevated exhaled nitric oxide levels. Airway inflammation using sputum cell counts has not been assessed in obese patients with airway diseases. OBJECTIVE: The primary aim of this study was to determine whether obesity (based on BMI) is associated with eosinophilic or neutrophilic bronchitis. METHODS: The results from a database of induced sputum cell counts were compared with BMI and analysed using correlation statistics, regression and parametric and non-parametric analysis. RESULTS: Seven-hundred and twenty-seven adult participants with an equal number of sputum samples were included in the analysis. BMI varied from 14.5 to 55 kg/m(2). Sputum total cell count (mean+/-SD: 12.9 x 10(6) cell/g+/-21.5), eosinophil percent (median; min to max: 0.3%; 0-89.0), and neutrophil percent (mean+/-SD: 63.5+/-26.6%) were within normal limits. Participants with asthma had a higher percentage of sputum eosinophils than those without asthma (P=0.01). However, there was no difference in the total or differential cell counts among the obese and non-obese participants, when the data were analysed according to BMI category, gender, dose of inhaled corticosteroid, and presence or absence of asthma. CONCLUSION: In this large sample of adult asthmatic and non-asthmatic participants, there was no association between BMI and airway inflammation measured by sputum cell counts. Other mechanisms to explain the relationship between obesity and asthma will need to be explored if this association is to be better understood.

Patient education materials for mental health problems in family practice: does location matter?

INTRODUCTION: This study examines how frequently family physicians display printed educational materials about mental health problems in their practices and where these materials should be located for optimal effect. METHODS: A cross-sectional observational study of pamphlet display practices in 13 family physicians' offices in Hamilton, Ontario, Canada was followed by an intervention which placed selected mental health educational materials in waiting rooms and examining rooms, and monitored the pick up rate from each location by patients. MAIN RESULTS: The study found that few mental health pamphlets were displayed by the participating physicians, that when a range of these pamphlets was made available, pamphlets on mood disorders were the most popular, and that significantly more pamphlets were picked up from examining rooms than from waiting rooms. CONCLUSIONS: We conclude that patients are interested in having access to printed materials about mental health problems, and that the optimal location is in display racks in examining rooms.

Does laparoscopic fundoplication provide long-term control of gastroesophageal reflux related cough?

BACKGROUND: Of patients with chronic cough, 21% have GERD. Up to half of these patients may not respond adequately to medical, but the long-term results of antireflux surgery for cough is unknown. METHODS: A total of 905 patients (209 with respiratory symptoms, mainly cough) underwent laparoscopic Nissen fundoplication. Preoperatively patients underwent esophageal motility studies, 24-h pH monitoring, and symptom evaluation using a validated scale. Of eligible patients, 81% were followed at 6 months, 73% at 2 years, and 60% at 5 years. RESULTS: Before surgery, 83% of respiratory patients (RP) and 51% of nonrespiratory patients (NRP) had cough. RP had higher cough scores ( p < 0.0001), but improvement in cough compared to baseline was similar in the RP and NRP ( p = 0.1105 at 6 months, 0.4206 at 2 years, and 0.1348 at 5 years). Cough improved in 83% at 6 months, 74% at 2 years, and 71% at 5 years. CONCLUSIONS: Laparoscopic Nissen fundoplication is successful in the long-term control of GERD-related cough, even in patients who fail medical therapy.

Does laparoscopic antireflux surgery prevent the occurrence of transient lower esophageal sphincter relaxation?

BACKGROUND: Transient lower esophageal sphincter relaxation (TLESR) is the most common mechanism underlying gastroesophageal reflux disease (GERD), causing 70% to 100% of the reflux episodes in normal subjects and 63% to 74% of the reflux episodes in patients with reflux disease. This study aimed to evaluate the effect of laparoscopic Nissen fundoplication on TLESR in patients with proven GERD. METHODS: We prospectively followed 73 consecutive patients (13 men and 60 women; mean age, 43.7 +/- 1.72 years) with proven diagnosis of GERD and reported TLESRs found during a 40-min esophageal manometric study. These patients had repeat testing 6 months after undergoing laparoscopic Nissen fundoplication. RESULTS: Laparoscopic Nissen fundoplication increased the basal and nadir lower esophageal sphincter (LES) pressure and significantly reduced the number of TLESRs during the manometric study. No patients after surgery exhibited TLESR with nadir less than 2 mmHg. However, 8 of the 73 patients (11%) exhibited TLESR to a nadir exceeding 50% of basal pressure (mean nadir, 5.0 +/- 1.07 mmHg). CONCLUSIONS: The number of TLESRs is reduced significantly by antireflux surgery. Even accounting for increased basal and nadir pressures, the incidence of TLESR is reduced, suggesting that there may be additional mechanisms involved in this process.

Preoperative symptom evaluation and esophageal acid infusion predict response to laparoscopic Nissen fundoplication in gastroesophageal reflux patients who present with cough.

BACKGROUND: Most patients with cough and gastroesophageal reflux disease (GERD) improve on medical treatment with proton pump inhibitors (PPI). Nonresponders may be considered for antireflux surgery, but the selection of patients is difficult. METHODS: We have performed laparoscopic Nissen fundoplications (LNF) in 677 patients. Of these patients, 81% have undergone 6-month follow-up assessment with 24-h pH testing, esophageal manometry, symptom scores, and quality-of-life scores. RESULTS: LNF controlled heartburn in 93% and improved cough in 81%. Stepwise multiple regression showed that the preoperative cough score (r = 0.620, p <0.0001) and change in cough on and off PPI (r = 0.296, p = 0.0002) predicted improvement after surgery. A positive result on a randomized acid infusion test was associated with a greater improvement in cough after surgery (p = 0.0243). CONCLUSION: An acid infusion test and assessment of cough on and off PPI may be useful preoperative tools for the selection of patients with cough for LNF.

Sputum cell counts and exhaled nitric oxide in patients with gastroesophageal reflux, and cough or asthma.

BACKGROUND: Gastroesophageal reflux (GER) is commonly associated with chronic cough and asthma, but there is little or no information on the nature of any associated airway inflammation. OBJECTIVE: To observe whether the association with GER worsens airway inflammation in patients with chronic cough or asthma. PATIENTS AND METHODS: The airway inflammatory indexes in induced sputum and exhaled air were examined in a cross-sectional study of 11 patients with cough and GER, nine patients with mildly symptomatic asthma and GER, nine patients with mildly symptomatic asthma without GER and nine normal, healthy control subjects. GER was shown objectively by 24 h ambulatory pH recording. RESULTS: The sputum total cell count, the proportion of neutrophils and macrophages, and the fibrinogen level were normal in all four groups, with no significant differences among the groups. The sputum eosinophil and metachromatic cell percentages, and eosinophil cationic protein levels were normal in patients with cough and GER. They were significantly increased in patients with asthma compared with healthy subjects (P<0.01) and patients with cough (P<0.01), but were not different between groups with and without GER. Exhaled nitric oxide levels showed similar results (P<0.01). The correlations between the number of episodes of reflux and the proportion of sputum eosinophils, neutrophils or exhaled nitric oxide were modest but not significant. CONCLUSIONS: GER, when associated with cough or mildly symptomatic asthma, does not cause or aggravate existing airway inflammation as measured by induced sputum cell counts and fibrinogen level, or by exhaled nitric oxide.

Genetic variation in the recognition of Porphyromonas gingivalis antigens in mice.

T-cell cytokine profiles, anti-Porphyromonas gingivalis antibodies and Western blot analysis of antibody responses were examined in BALB/c, CBA/CaH, C57BL6 and DBA/2J mice immunized intraperitoneally with different doses of P. gingivalis outer membrane antigens. Splenic CD4 and CD8 cells were examined for intracytoplasmic interleukin (IL)-4, interferon (IFN)-gamma and IL-10 by FACS analysis and levels of anti-P. gingivalis antibodies in the serum samples determined by enzyme-linked immunosorbent assay. Western blot analysis was performed on the sera from mice immunized with 100 microg of P. gingivalis antigens. The four strains of mice demonstrated varying degrees of T-cell immunity, although the T-cell cytokine profiles exhibited by each strain were not affected by different immunizing doses. While BALB/c and DBA/2J mice exhibited responses that peaked at immunizing doses of 100-200 microg of P. gingivalis antigens, CBA/CaH and C57BL6 demonstrated weak T-cell responsiveness compared with control mice. Like the T-cell responses, serum antibody levels were not dose dependent. DBA/2J exhibited the lowest levels of anti-P. gingivalis antibodies followed by BALB/c with CBA/CaH and C57BL6 mice demonstrating the highest levels. Western blot analysis showed that there were differences in reactivity between the strains to a group of 13 antigens ranging in molecular weight from 15 to 43 kDa. Antibody responses to a number of these bands in BALB/c mice were of low density, whereas CBA/CaH and C57BL6 mice demonstrated high-density bands and DBA/2J mice showed medium to high responses. In conclusion, different immunizing doses of P. gingivalis outer membrane antigens had little effect on the T-cell cytokine responses and serum anti-P. gingivalis antibody levels. Western blot analysis, however, indicated that the four strains of mice exhibited different reactivity to some lower-molecular-weight antigens. Future studies are required to determine the significance of these differences, which may affect the outcome of P. gingivalis infection.

Noninvasive methods for quantitative assessment of transfusional iron overload in sickle cell disease.

Because optimal management of iron chelation therapy in patients with sickle cell disease and transfusional iron overload requires accurate determination of the magnitude of iron excess, a variety of techniques for evaluating iron overload are under development, including measurement of serum ferritin iron levels, x-ray fluorescence of iron, magnetic resonance imaging, computed tomography, and measurement of magnetic susceptibility. The most promising methods for noninvasive assessment of body iron stores in patients with sickle cell anemia and transfusional iron overload are based on measurement of hepatic magnetic susceptibility, either using superconducting quantum interference device (SQUID) susceptometry or, potentially, magnetic resonance susceptometry.

Identification of HLA-A*2306.

We describe a novel allele encoding HLA-A23: A*2306, discovered in an African-American individual, whose DNA was HLA typed as part of a quality control exercise. Direct sequencing typing identified A*2301 and A*6601 with an unexpected heterozygous peak at position 331. As position 331 is at the end of exon 2, near the priming site for the B3.6 anti-sense sequencing primer, the sequencing data is not optimal in this region and sequencing from the sense primer is relied on. In addition the new polymorphism was not at an expected polymorphic position and could easily have been missed, leading to the assignment of A*2301. However, data from reference strand mediated conformation analysis showed distinct new mobilities from those expected for A*2301 with two different fluorescent-labelled references, leading to the conclusion that the heterozygous peak seen at position 331 was a true variant of the A*2301 allele. A*2306 is most similar to A*2301 with 1 nucleotide difference at position 331 in exon 2 which was previously a conserved position. This mutation results in an amino acid substitution of glutamine for glutamate at residue 87.

Notification of infectious diseases by general practitioners: a quantitative and qualitative study.

OBJECTIVE: To examine general practitioners' knowledge and practices concerning reporting of notifiable conditions, and to examine barriers to notification of infectious diseases by general practitioners and identify strategies for improving the notification process. DESIGN AND SETTING: Audit of the 100 most recent notifications received by the South Eastern Sydney Public Health Unit of cases of each of hepatitis A, pertussis and measles; and focus groups with GPs practising in Sydney's eastern and southern suburbs, some of whom were selected on the basis of their notification practices. RESULTS: Although these diseases are notifiable on clinical suspicion, only about 40% of the hepatitis A and pertussis cases and 80% of measles cases (54% overall) had been notified by GPs. Delays between doctor and laboratory notifications were an average of seven days for hepatitis A, 19 days for pertussis and seven days for measles. Focus groups showed that at least some GPs have poor understanding of the process of notification, most felt uncomfortable notifying an unconfirmed case, many preferred to leave notification to the laboratory because of concerns about damaging the doctor-patient relationship, and that there is need for financial or other incentives. CONCLUSIONS: There are deficiencies in the completeness and timeliness of notification by GPs which may adversely affect the timing of prophylaxis and outbreak control. Notification by GPs may be improved by such strategies as better notification forms and better feedback to doctors on the outcomes.

Reproducibility, validity, and responsiveness of a disease-specific symptom questionnaire for gastroesophageal reflux disease.

The purpose of this study was to establish the reproducibility, validity, and responsiveness of a symptom questionnaire to assess patients with gastroesophageal reflux disease (GERD). A total of 300 patients with GERD completed questionnaires before and 6 months after laparoscopic Nissen fundoplication. Forty-six GERD patients who continued on omeprazole served as controls. Lower esophageal sphincter pressure, 24-h pH, and quality of life (SF36) were measured at baseline and follow-up. Reproducibility was calculated as an intraclass correlation coefficient (ICC) from a repeated-measures analysis of variance on symptom scores (SS) on two consecutive days. Validity was established by correlating SS with 24-h pH and SF36 scores. Responsiveness was calculated as the the ratio of the mean paired difference in score in the surgical group to the within-subject variability in control subjects. Reproducibility was very high, as revealed by an ICC of 0.92. Strong correlations between SS and SF36 scores at baseline and after surgery demonstrated high cross-sectional validity. Correlation between change in SS and change in pH, SF36 pain, general health, and physical health scores demonstrated longitudinal validity. The mean (95% confidence interval) paired differences in SS were 25.6 (23.7, 27.5) in the study and 2.0 (-3.2, 7.3) in the control groups, and the responsive index was 1.0. The estimated minimally important clinical difference was 7. We conclude that the symptom score is a reproducible, valid, and responsive instrument for assessing symptoms caused by GERD.

Lipid-laden macrophages in induced sputum are a marker of oropharyngeal reflux and possible gastric aspiration.

The diagnostic properties of a "lipid index" of macrophages in induced sputum as a noninvasive marker of aspiration of acidic gastric contents were evaluated. In a cross-sectional study, 33 subjects (17 with symptoms suggestive of gastrooesophageal reflux) with normal chest radiographs and no symptoms of aspiration or sinus disease, underwent dual-channel 24-h ambulatory oesophageal pH recording and sputum induction. Oropharyngeal reflux, defined as at least one episode of a fall in pH to <4 at the upper oesophageal electrode, was considered indicative of aspiration of acidic gastric contents ("gold standard"). An index for the presence of intracellular lipid in sputum macrophages, detected by oil red O stain, was obtained. The sensitivity, specificity and predictive values of this "lipid index" were calculated. The "lipid index" could be calculated in 29 of 33 samples with high interobserver repeatability (intraclass correlation coefficient 0.96). Twenty subjects showed oropharyngeal reflux and nine did not. The median "lipid index" in subjects with oropharyngeal reflux (24.5) was significantly greater than that in those without reflux (1.0) (p<0.001). A "lipid index" of 7.0 had a sensitivity of 90%, a specificity of 89%, a positive predictive value of 95% and a negative predictive value of 80%. A "lipid index" of 7.0 in the macrophages of induced sputum is a good marker of oropharyngeal reflux.

Gastro-oesophageal reflux related cough and its response to laparoscopic fundoplication.

BACKGROUND: This study was designed to determine prospectively the rate of cough before and after laparoscopic Nissen fundoplication performed for the control of gastro-oesophageal reflux disease. METHODS: One hundred and ninety five consecutive patients (76 men) of mean (SD) age 46.9 (14.1) years with proven gastro-oesophageal reflux disease, who were either on long term omeprazole (n = 187) or who had not responded to a trial of omeprazole (n = 8), took part in the study which was carried out in a university teaching hospital that included a regional respiratory referral centre. Patients underwent oesophageal manometry, 24 hour oesophageal pH testing, and symptom score evaluation by an independent observer before and six months after laparoscopic Nissen fundoplication. RESULTS: One hundred and thirty three patients presented with reflux symptoms and 62 with respiratory symptoms; 68% of patients complained of cough before surgery (86% with respiratory symptoms, 60% with gastrointestinal symptoms). The percentage reflux time in 24 hours fell significantly (p < 0.0001) from a mean (SD) of 9.38 (10.99)% to 1.22 (2.92)%, lower oesophageal sphincter tone rose significantly (p < 0.0001) from a mean (SD) of 7.71 (5.90) mm Hg to 21.74 (10.84) mm Hg, and the cough score fell from a median value of 8.0 (IQR 12.0) to 0 (IQR 3) following surgery. Of the patients with cough, 51% were cough free after surgery and 31% improved. The patients with respiratory symptoms had a higher cough score before (median 12.0 (IQR 5.5) versus 4.0 (IQR 8.75), p < 0.0001) and after surgery (median 1 (7.5) versus 0.0 (IQR 1.0), p = 0.0045) than those with gastrointestinal symptoms. CONCLUSIONS: Patients who present to gastroenterologists with severe reflux commonly complain of cough. Laparoscopic Nissen fundoplication is effective in the control of cough in patients with gastro-oesophageal reflux disease, with or without primary respiratory disease.

Prospective evaluation of dysphagia before and after laparoscopic Nissen fundoplication without routine division of short gastrics.

The incidence and severity of dysphagia before and 6 months after laparoscopic Nissen fundoplication without routine division of short gastric vessels are presented. Laparoscopic Nissen fundoplication was undertaken in 195 patients over 32 months with 116 patients who had prospective follow-up longer than 6 months. Patients underwent a 24-h pH recording, esophageal manometry, and symptom score assessment before and 6 months after surgery. There was a significant (p < 0.0001) improvement in the percent of reflux in 24-h (8.61 +/- 0.74 to 0.68 +/- 0.12), lower esophageal pressure (8.53 +/- 0.51 to 23.11 +/- 1.1 mm Hg), and reflux symptom scores (40.97 +/- 1.13 to 12.11 +/- 1.1) at 6 months. A similar improvement (p < 0.0001) was also observed in the dysphagia symptom score (4.58 +/- 0.38 to 1.96 +/- 0.32), with more than half the patients reporting improvement after surgery. No correlation was observed between the change in dysphagia score and the postoperative lower esophageal pressure or esophageal motor function. These data suggest that the incidence of clinically significant dysphagia after laparoscopic Nissen fundoplication, even without division of short gastric vessels, is low. Improvement in the dysphagia score after surgery is interesting and warrants further investigation.