At the same time, we compared these biological behaviors with tra

At the same time, we compared these biological behaviors with traditional endothelial cell, human umbilical vein endothelial Selleck P505-15 cell (HUVEC) and the original cancer cells. Further, we tried to explore the underlying mechanisms

by detection the expression of some relative genes. Methods Cell culture Human epithelial ovarian carcinoma cell lines SKOV-3 and ES-2 were purchased from American Type Culture Collection (ATCC, Manassas, VA), and were maintained in McCoy’s 5a. Primary human umbilical vein endothelial cells (HUVEC) were isolated from umbilical vein and cultured as described previously [14] Three-dimensional cultures and hypoxic treatment Thirty microliters of Matrigel (B&D, Bedford, MA) were dropped onto each glass coverslip in a 12-well culture plate and polymerized for 1 h at room

temperature, followed by 30 min’s incubation at 37°C in a humidified 5% CO2 incubator, as described previously [15]. Tumor cells (1 × 104) were seeded onto the three-dimensional gel. The medium supplied with 15% FBS was changed every 36 h. Hypoxic condition was created by flushing 5% CO2 and 95% N2 through a modified chamber (Mitsubishi, Japan), until O2 concentration was reduced to 1%, measured with a Mini oxygen meter. The culture system was sealed and incubated at 37°C [16]. The cells were treated with 50 nM Sirolimus (Sigma, St. Louis, MO) in DMSO to inhibit the role of HIF-1α under hypoxia when necessary. Proliferation assay For the proliferation assay, 1 × 104 SKOV-3, ES-2 and HUVEC cells, were seeded into a flat bottom 96-well

plate and incubated at 37°C for 3 and 7 d under normoxia or hypoxia (1% O2) respectively, prior to the addition of 20 μL of MTT solution (5 mg/ml in PBS). After incubated for additional 4 h at 37°C, absorbance at 490 nm was measured with a multi-function reader (Tecan GENios, Zurich, Switzerland) to determine cell viability. Cell cycle and apoptosis assay Cell cycle and apoptosis assay were performed on cells with or without hypoxia treatment (for 3 or 7 d) to determine whether hypoxia regulates the growth phase and apoptosis of epithelial ovarian cells. Cells were MYO10 trypsinized and centrifuged at 300 × g (1000 rpm) for 5 min, then resuspended (1 × 106 cells/ml) and fixed with 70% ice-cold ethanol for 30 min, followed by centrifuged, washed and resuspended in 500 μl PBS contained 10 μl of DNase free RNase (final concentration is 1‰). After 30 min incubation, pyridine iodide (PI, 0.05 mg/ml) was added to the solution to incubate for an additional 15 min in the dark and filtered by a nylon mesh to remove cell clusters. The fluorescence of PI was measured using FACS Calibur Flow Cytometer (Becton-Dickinson, San Jose, CA). Cell subpopulations in G0/G1, S and G2/M phases and apoptosis were calculated by gating analysis based on differences in DNA content.

For the films with 13% and 21% Cu (c and e), the dealloying proce

For the films with 13% and 21% Cu (c and e), the dealloying procedure decreased the copper

content in the film and resulted in surface pits where copper was removed (d and f). The pits formed in the sample with the smaller initial Cu concentration (d) are smaller than those formed in the sample with the larger initial Cu concentration (f). This can be seen more clearly in the higher resolution SEM images of the post-dealloyed films in Figure 4. Figure 3 SEM images of NiCu films before (a, c, e) and after (b, d, f) the dealloying procedure. The initial copper content in the films are (a) 9.0±0.5%, AZD5582 molecular weight (c) 12.6±0.6%, and (e) 21.4±1.1%. The copper content in the dealloyed films are (b) 9.5±0.5%, (d) 11.4±0.6%, and (f) 13.9±0.7%. The scale bar is 5 μm for all the images. Figure 4 Higher resolution SEM images of the dealloyed NiCu films in (a) Figure 3 d find more and (b) Figure 3 f. The scale bar is 1 μm for both images. To compare the resulting electrochemically accessible surface areas of the samples, the electrochemical double-layer capacitance was measured for each sample both before and after

the dealloying step. In the simplest model, this capacitance is proportional to the surface area of the sample accessible via electrochemistry and thus provides a semi-quantitative measure of that area. Figure 5 shows the ratio of the measured capacitance after the dealloying step to before the dealloying step as a function of the amount of copper selectively removed. In the figure, negative Cu removed indicates that Ni was

selectively removed in the dealloying step; for these samples, when the uncertainties are taken into account, the Cu removed amounts are statistically equivalent to zero. The dashed line indicates identical measured capacitances before and after dealloying. Figure 5 Ratio of measured capacitance after to before the dealloying step. The capacitance ratio as a function of the copper composition (at.%) removed in the dealloying step. Negative Cu removed indicates that Ni was selectively removed in the dealloying step rather than Cu. The dashed line indicates identical measured capacitances before and after Tolmetin dealloying. For all the samples studied, the capacitance either stayed statistically the same or increased, suggesting that the dealloying procedure either did not change the effective surface area of the sample or caused it to increase. For the samples with between 3% and 15% Cu removed, the capacitance ratio decreases as the amount of copper removed increases. This observation is consistent with the SEM images in Figures 3 and 4. The samples with larger initial copper content tended to have rougher initial topography, such as that in Figure 3e, and thus had higher initial capacitance measurements. In addition, those samples tended to have larger pits seen in the post-dealloy topography, such as in Figure 3f, which increased the measured capacitance only modestly.

5) supplemented with 0 5 ml of 0 25 g/ml TMAO solution The resul

5) supplemented with 0.5 ml of 0.25 g/ml TMAO solution. The resulting clear bands on the blue background indicate the presence of active TMAO reductase in the gel. Growth assessment of strains in M9-TMAO media The overnight cultures of different tat genes deletion mutants and complement strains (listed in Table 1) and the wild type strain N16961 were diluted 1:100 and incubated in fresh LB MK-2206 solubility dmso to OD600 more than 0.8. Then the culture of each was adjusted with LB to OD600 of 0.8. Then they were then diluted 1:100, and 50 μl of each culture was transferred into M9-TMAO media and subsequently cultured statically at 37°C in the anaerobic jar (Oxoid). The vacuum extractor was used to extract the air in the anaerobic

jar to lower atmospheric pressure (-10 millimeters of mercury), and then H2 and CO2 were inflated to normal atmospheric pressure. The culture was grown for 24 h, and then the OD600 of each culture was determined. Pritelivir Motility assay Motility of N16961 and N169-dtatABC cells was tested on 0.3% minimal motility agar containing 1% peptone and 0.5% NaCl (wt/v). Briefly, cell cultures grown in LB broth overnight at 37°C were diluted 1:1000. Cell cultures were then grown to OD600 0.2. Subsequently, each strain was inoculated

onto the surface of the motility U type tubes. Motility was examined after 12 h and 16 h of incubation at 37°C. The percentage of the length of growth diffusion in the agar of the mutant strain N169-dtatABC compared to the wild type strain was calculated. At least five independent motility assays were carried out for each strain and condition. Outer membrane integrity assay We detected the outer membrane

integrity Rebamipide according to the method of reference [26]. The wild type strain N16961 and the Tat mutant strain N169-dtatABC were cultured overnight and then diluted 1:100 into fresh LB and grown to OD600 0.4. Five milliliters of fresh LB was added into each tube, and SDS or Gentamicin (Get) was added to final concentrations of 0 to 2.5% and 0 to 500 μg/ml, respectively. Experiments were performed in triplicate for N16961 and Get. After SDS or Get addition, all tubes were grown at 37°C for 3 h at 250 rpm, after which the OD600 of each culture tube was measured. We defined the OD600 of the wild type strain cultured in LB without SDS and Get as 100%. The OD600 values of the other conditions were converted to the percentage of OD600 of the wild type strain cultured in LB without SDS and Get. To determine whether the outer membrane of the mutants was destroyed, the results are plotted as SDS or Get dilution on the X-axis and OD600 percentage on the Y-axis [26]. Flagellum extraction and quantification Bacterial cells were recovered from a 600 ml LB culture of N16961 and N169-dtatABC incubated overnight at 37°C and then centrifuged for 5 min at 10,000 g. The pellets were resuspended in PBS buffer and vortexed for 5 min, with a 30 s interval after 2.5 min.

This cell suspension constituted the

standard starting in

This cell suspension constituted the

standard starting inoculum (S) as defined by CLSI guidelines for antimicrobial susceptibility testing [68]. Double (D) and half (H) the size of the standard inoculum were used to evaluate the effect of the initial cell AZD3965 clinical trial density on the activity of biocides towards S. algae. To check the actual starting cell number, a 200 μl sample of the inoculum was serially tenfold diluted from 10−1 to 10−8. Four 10 μl drops from each dilution were spotted on agar plates and incubated. Colony formation was assessed after 24 h. Microscopy: general procedures For microscopy experiments, the bottoms of the wells of a microtiter plate were mechanically sectioned with a computer numerical control milling machine (Fagor CNC 8055 M) in order to use exactly the same substrate as in previous tests. The sectioned discs thus obtained (5.86-5.98 mm in diameter, 1.00-1.08 mm in height, data from 15 random

PLX-4720 supplier measurements) were carefully disengaged and sterilised by a brief sonication in ethanol and UV irradiation before their use in the experiments. To develop the biofilms, the discs were placed at the bottom of a 24-well microtiter plate. Two-mililiter bacterial cultures were prepared in the appropriate medium following the same procedures as described previously. After the incubation period, discs were rinsed three times with FSW and kept immersed upon their use in the microscope. Confocal Laser Scanning Microscopy Biofilms formed on polystyrene discs were fluorescently stained with acridine orange (AO), a membrane permeant nucleic acid stain that intercalates dsDNA and binds to ssDNA as well as to ssRNA through dye-base stacking to give broad spectrum fluorescence when excited Ribose-5-phosphate isomerase at 476 nm [69]. This compound stains all cells in a biofilm, live or dead, and may

also bind to nucleic acids that are present in the extracellular matrix. To stain biofilms, discs were immersed in 0.1% w/v AO (Sigma-Aldrich) in PBS for 5 min at room temperature and washed with FSW. Fluorescently labelled biofilms were placed in two drops of 0.9% FSW on the surface of a glass coverslip and were examined using an Olympus Fluoview 1000 Confocal Laser Scanning Microscope. Each biofilm was scanned at 4 positions randomly selected at the microscope stage and confocal image series were generated by optical sectioning at each of these positions. Three independent biofilm experiments were performed, and image stacks of 512×512 pixels were collected for quantification. Image combining and processing were performed with the Imaris software package, version 4.0 (Bitplane AG, Zürich, Switzerland). The biofilm structure was quantified using the software program COMSTAT [70] available as free downloadable software at http://​www.​imageanalysis.​dk. COMSTAT converts pixels from confocal image stacks into numerical values, facilitating quantitative characterization of each structural component within 3D biofilm images [71].

(1998) and Laestadius et al (2008) Furthermore, it has to be no

(1998) and Laestadius et al. (2008). Furthermore, it has to be noted that we used as reference the scores from a working selleck chemicals llc population in Germany to study functional impairment. There might be differences between the Dutch and the German population with respect to this issue, but we do not have indications for that. Aublet-Cuvelier et al. (2006) performed a follow-up study on the course of work-related upper extremity disorders during three consecutive years at a household appliance assembly company (n = 459). They found a relatively stable annual prevalence of 20–24% and a high annual incidence

(9.8–13.5%) of cases and of annual recoveries (37.0–44.3%). The number of annual recoveries compares well with the favourable course in our study. Feleus et al. (2007)

reported that 42% of a working population (n = 473) with non-traumatic complaints of the arm, neck and shoulder still reported complaints after 6 months. This compares to our finding that complaints had decreased in 33% of the patients after 6 months of follow-up. Cheng et al. (2002) found significant improvements in the SF-36 physical functioning and bodily pain scores after a physical therapy (PT) intervention, but noted a variation in outcomes across injury regions. Patients with elbow disorders needed more physiotherapy care and did not improve in the SF-36 physical role domain compared to shoulder and Selleckchem MAPK Inhibitor Library wrist/hand groups (Cole and Hudak 1996). We concluded that the results of several studies on the course C1GALT1 of work-related upper

extremity disorders seem to be generally comparable to our findings. An interesting finding in our study was that the average VAS score of the general quality of life did not change, but the VAS quality of life scores with respect to health did increase. This might indicate that the functionality of the upper extremity does not have a major contribution to general quality of life. Reitsma (1999) considered the possibility of follow-up studies linked to registries. He concluded that in most registries follow-up or historical information is not recorded, is short term or is missing and that the role of registries can be extended by creating longitudinal data. This can be done either by record linkage of existing data or by sample projects. This type of information is important in order to set priorities for preventive policy and to monitor the effects of policy interventions. The impact of diseases in terms of severity and duration has to be taken into account in policy making. Furthermore, trends can be monitored not only on the incidence of diseases but also on their course and consequences. If appropriate data can be obtained, the monitoring of economic costs could be added to the set of monitoring instruments. Further research can be performed on the use of registries and related sample projects for preventive policy.

[34] and Malm et al [35] was used to induce the eccentric muscle

[34] and Malm et al. [35] was used to induce the eccentric muscle injury. After a 10-min warm-up at a speed chosen by the subject, subjects ran downhill (treadmill grade -10%) at a constant speed for 45 minutes. Running speed during the 45-min exercise was to be maintained at the anaerobic threshold, which was determined prior

to the test by measurement of lactate concentration in capillary blood during a 5-min run at a treadmill inclination Ro 61-8048 ic50 of 3%. A speed corresponding to a lactate concentration of 3.5-5 mmol/L was considered appropriate and therefore maintained throughout the exercise protocol. Subjects performed ten-min exercise bouts during the week prior to the study day (days -7 and -5) to familiarize with the exercise protocol and to break down more susceptible muscle fibres, in order to achieve similar

fibre composition and standardize the baseline level in all subject [36, 37]. One hour before the eccentric injury protocol all subjects received an oral nutritional supplement containing 25 to 30 g of carbohydrates and 2–4 g of protein. Also, hydration was assured by consumption of approximately 500 mL of mineral water from 30 min. prior to the start of the test. Subjects were allowed to drink water during the test. Magnetic resonance CX 5461 imaging (MRI) A high magnetic field system was used (Signa 1.5 T, G.E. Milwaukee, WI, USA). Images were acquired 48 hours after exercise, with the subjects in the supine decubitus position. Both thighs were explored. The diagnosis was based on MRI signal alterations in any muscular group both in the flexor and the extensor compartment, as well as on signal asymmetry as compared with the contralateral homonymous muscular group. The radiologist was blinded to the treatment group. Five non-contiguous axial

PRKD3 imaging slices (2-mm thickness, 2-mm gap) were selected. In order to quantify muscle injury, each thigh was divided into three compartments (anterior, posterior, medial) (Figure 1). A compartment was considered positive for muscular injury when an area of high signal intensity on T2-weighted and STIR sequences was observed in at least one muscle. Figure 1 STIR sectional image of both thighs in the middle third. Asterisk marks muscle area with increased uptake. Muscle biopsies Muscle biopsies were performed 48 hours after exercise to obtain samples for the analysis of markers of cellular injury (muscle myeloperoxidase [MPO] activity, immunohistochemical analysis of albumin [38] and CD3 positive cells). A skin incision was performed with a 5 mm blade. The same skin incision was used for both muscle biopsies, changing the needle direction [34, 39] . Two biopsies were carried out from the middle third of each vastus lateralis, under ultrasound control. Muscle samples were obtained using a Vacora System Biopsy gun (Bard Medical Systems, Tempe, AZ, USA), with a coaxial needle of 10G × 140 mm.

It has also been suggested that the two components of this partic

It has also been suggested that the two components of this particular regulatory system do not always act in tandem specifically in response to acid stress. From the results obtained in this study, we cannot speculate on the overexpression of CpxA in PA adapted cultures-as CpxA is a membrane localized protein and this study focused on soluble proteins. It may be informative, however, to examine the expression profile of CpxA in PA adapted cultures in order to decipher if CpxR works in a concerted manner with CpxA to protect cells from acid stress following the onset of PA-induced acid resistance. Conclusion

It is apparent that long this website term PA adaptation of S. Enteritidis is associated with differential protein expression, with the synthesis of

certain proteins being significantly upregulated. Sapitinib Of these proteins, Dps and CpxR are those commonly associated with virulence and we have not only demonstrated that they are inducible by PA, but also that they are crucial for PA-induced acid resistance in S. Enteritidis. These results clearly demonstrate that Dps and CpxR play an important role in PA-induced acid resistance. It is also apparent that overexpression of either Dps or CpxR alone in PA adapted cultures is not sufficient to confer increased acid resistance. Acknowledgements This study was supported by a USDA Food Safety Consortium grant. Electronic supplementary material

Additional file 1: Protein Report C. Mass spectrometry report for RplE (PDF 370 KB) Additional file 2: Protein Report B. Mass spectrometry report for RplF (PDF 262 KB) Additional file 3: Protein Report A. Mass spectrometry report for SodA (PDF 343 KB) Additional file 4: Protein Report D. Mass spectrometry report for CpxR and Dps (PDF 345 KB) References 1. Callaway TR, Edrington TS, Anderson RC, Byrd JA, Nisbet DJ: Gastrointestinal microbial ecology and the safety of our food supply as related to Salmonella . J Anim Sci 2008,86(E suppl):E163-E172.PubMed 2. Foster JW, Hall HK: Adaptive Acidification Cepharanthine Tolerance Response of Salmonella typhimurium . J Bacteriol 1990, 172:771–778.PubMed 3. Lee IS, Slonczewski JL, Foster JW: A Low-pH-Inducible, Stationary-Phase Acid Tolerance Response in Salmonella typhimurium . J Bacteriol 1994, 176:1422–1426.PubMed 4. Lin J, Lee IS, Frey J, Slonczewski JL, Foster JW: Comparative Analysis of Extreme Acid Survival in Salmonella typhimurium , Shigella flexneri , and Escherichia coli . J Bacteriol 1995, 177:4097–4104.PubMed 5. Kwon YM, Ricke SC: Induction of acid resistance of Salmonella typhimurium by exposure to short-chain fatty acids. Appl Environ Microbiol 1998, 64:3458–3463.PubMed 6. Gahan CG, Hill C: The relationship between acid stress response and virulence in Salmonella typhimurium and Listeria monocytogenes . Int J Food Microbiol 1999, 50:90–100.CrossRef 7.

Either 5 or 10 μL of the supernatant was injected for tissue or p

Either 5 or 10 μL of the supernatant was injected for tissue or plasma samples, respectively. Calibration curves and QC samples were prepared

in both brain and liver, for tissue sample analysis. The Selleckchem JNK inhibitor working ranges for liver and brain were 0.125–100 and 0.125–25 ng/mL, respectively. Equipment High performance liquid chromatography was carried out on an Agilent 1100 system (Agilent Technology, Palo Alto, CA), coupled with a single-quadrupole mass spectrometer, utilizing electrospray ionization in positive mode. Samples were cooled to 4°C in a thermostated autosampler and the column compartment, containing a Waters SymmetryShield RP8 column (2.1 × 50 mm, 3.5 μm), was maintained at 35°C. Samples were eluted using a gradient mobile phase, comprised of 10 mM ammonium acetate with 0.1% formic acid and methanol, running at a flow rate of 0.35 mL/min for 10 min, including re-equilibration. Mass spectrometric conditions were as follows: fragmentor, 150 V; gain, 2; drying gas flow, 10 L/min; drying gas temperature, 300°C; nebulizer pressure, 40 OSI-906 molecular weight psi; and capillary voltage, 1500 V. Selected-ion monitoring

was accomplished at m/z 494.2 for imatinib and m/z 213.1 for the internal standard. The chromatographic data were acquired and analyzed using the Chemstation software package (Agilent). Validation procedures Calculation of accuracy and precision was carried out according to procedures reported in detail previously [17]. Calibration samples were prepared fresh each

day in the relevant matrix and frozen QC samples were defrosted and analyzed. A 1/x2 weighting scheme was employed in the generation of standard curves to account for concentration dependent variance. Detector response for plasma was found to be linear in the imatinib concentration range of 10–1000 ng/mL. Plasma accuracy and precision were evaluated with QC samples. Overall, the assay was found to be accurate (deviation of less than 10% for QCs) and precise (within run precision <10%, between run precision <12.6%) for plasma, liver, and brain. Animals All experiments were performed on six-week old, male, Fludarabine order Balb/C mice obtained from Charles River Laboratories (Wilmington, MA). The mice weighed approximately 15 to 20 g at the time of study. All mice were allowed unlimited access to water and rodent chow prior to, and during the experiment. Blank mouse liver and brain samples were harvested from surplus mice following euthanasia. NCI-Frederick is accredited by AAALAC International and follows the Public Health Service Policy for the Care and Use of Laboratory Animals. Animal care was provided in accordance with the procedures outlined in the “”Guide for Care and Use of Laboratory Animals”" (National Research Council; 1996; National Academy Press; Washington, DC). The study design and protocol were approved by the NCI Animal Care and Use Committee (Bethesda, MD).

Visible biofilm remained after draining the tubing for the refere

Visible biofilm remained after draining the tubing for the reference strain (DAY286) and the hwp1/hwp1 mutant, while no visible biofilm remained for the bcr1/bcr1 mutant. There was some residual NVP-BSK805 datasheet biofilm left after draining the tubing colonized by the als3/als3 mutant (before the ethanol rinse steps), but the adhesion to the surface was clearly much less than the reference strain. SEM images of the tubing

in the second row indicated that multilayer biofilm remained on the surface of the tubing for the reference strain and the hpw1/hpw1 mutant, while very few cells could be found for the bcr1/bcr1 and als3/als3 mutants. The most heavily colonized regions that were found are shown. (The ethanol dehydration removed all visible biofilm from the tubing for bcr1/bcr1 and als3/als3 mutant strains). Comparison of the firmly and loosely attached biofilm suggests that glycosylation, vesicle trafficking and transport contribute to the adhesive phenotype As shown in Figure (2d and 2e) a visible multilayered biofilm structure withstands MEK inhibitor the substantial shear force applied by draining the tubing for biofilms cultured for 1 h. A portion of the 1 h biofilms is typically removed from the surface

by this procedure. These two subpopulations are referred to as the 1 h firmly (1h F) and 1 h loosely (1h L) attached biofilm. We reasoned that comparing the transcriptional profiles of these two subpopulations might uncover genes that were subsequently differentially regulated to mediate detachment in our flow model. The comparison of 1h F and 1h L biofilms revealed 22 upregulated and 3 repressed transcripts (see Additional file 1). Upregulated genes fell into process ontological categories of vesicular trafficking, glycosylation

and transport. RT-qPCR confirmed Fenbendazole the changes in transcript levels of some genes enriched in glycosylation and vesicle trafficking functions that exhibited relatively small fold changes (Table 2). The distinct pattern of expression of these genes within the context of the time course analysis is discussed in the next section. Table 2 Genes up regulated in the 1hF/1hL comparison Gene Orf Microarray1 RT Q-PCR2 Vesicular trafficking SSS1 orf19.6828.1 1.56 1.63 ± 0.01 ERV29 orf19.4579 1.60 3.73 ± 0.41 SEC22 orf19.479.2 1.44 2.24 ± 0.1 EMP24 orf19.6293 1.44 1.24 ± 0.1 CHS7 orf19.2444 1.44 1.65 ± 0.12 YOP1 orf19.2168.3 1.55 1.67 ± 0.15 Glycosylation PMT4 orf19.4109 1.63 ND3 DPM2 orf19.1203.1 1.61 2.33 ± 0.11 DPM3 orf19.4600.1 1.48 2.12 ± 0.2 WBP1 orf19.2298 1.44 4.75 ± 0.11 Transport ADP1 orf19.459 1.68 ND CTR1 orf19.3646 1.54 ND ADY2 orf19.1224 1.69 ND TNA1 orf19.2397 1.68 ND ALP1 orf19.2337 1.58 ND 1Average fold change 2Log2 ratios. Each value is the mean ± standard deviation of two independent experiments each with three replicates.

​pdf [Accessed 2011 Dec 13] 60 European Medicines Agency Withd

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Sciences Working Group. Introductory guide for Standardised MedDRA Queries (SMQs) Version 13.0. Chantilly (VA): MedDRA Maintenance and Support Services Organization, 2010. 64. International Conference on Harmonisation of Technical Requirements for Registration Crenolanib mouse of Pharmaceuticals for Human Use. ICH harmonized tripartite guideline. Statistical principles for clinical trials: E9 [online]. Available from URL: http://​www.​ich.​org/​fileadmin/​Public_​Web_​Site/​ICH_​Products/​Guidelines/​Efficacy/​E9/​Step4/​E9_​Guideline.​pdf Branched chain aminotransferase [Accessed 2012 Jan 28]. 65. Greenland S, Robins JM. Estimation of a common effect parameter from sparse follow-up data. Biometrics 1985; 41 (1): 55–68.PubMedCrossRef 66. Miravitlles M. Moxifloxacin in respiratory tract infections. Expert Opin Pharmacother 2005; 6 (2): 283–93.PubMedCrossRef 67. Craig WA. Overview of newer antimicrobial

formulations for overcoming pneumococcal resistance. Am J Med 2004; 117 Suppl. 3A: 16S–22S.PubMed 68. File TM, Garau J, Jacobs MR, et al. Efficacy of a new pharmacokinetically enhanced formulation of amoxicillin/clavulanate (2000/125mg) in adults with community-acquired pneumonia caused by Streptococcus pneumoniae, including penicillin-resistant strains. Int J Antimicrob Agents 2005; 25 (2): 110–9.PubMedCrossRef 69. Aspa J, Rajas O, de Castro FR. Pneumococcal antimicrobial resistance: therapeutic strategy and management in community-acquired pneumonia. Expert Opin Pharmacother 2008; 9 (2): 229–41.PubMedCrossRef 70. Croom KF, Goa KL. Levofloxacin: a review of its use in the treatment of bacterial infections in the United States. Drugs 2003; 63 (24): 2769–802.PubMedCrossRef 71. Klugman KP. Bacteriological evidence of antibiotic failure in pneumococcal lower respiratory tract infections. Eur Respir J Suppl 2002; 36: 3s–8s.PubMedCrossRef 72. Odenholt I, Cars O.