Furthermore, the silica moiety of Fe3O4@SiO2-OCMCS-FA nanovehicle could be extended to fabricate mesoporous nanovehicle selleck chemicals llc which may increase surface area and pore volume. Thus, we believe that this strategy may provide a safe and efficient platform for antitumor drug delivery. Acknowledgements We gratefully acknowledge the assistance of Professor Zheng Xu from the State Key Laboratory of Coordination Chemistry in Nanjing University. The work was financially supported by the Fundamental Research Funds for the Central Universities (JKZD2013003). References 1. Shen JM, Yin T, Tian XZ, Gao FY, Xu S: Surface charge-switchable polymeric magnetic nanoparticles for the controlled release of anticancer

drug. ACS Appl Mater Interfaces 2013, 5:7014–7024.CrossRef 2. Lee JH, Lee K, Moon SH, Lee YH, Park TG, Cheon J: All-in-one target-cell-specific magnetic nanoparticles for simultaneous molecular imaging and siRNA delivery. Angew Chem Int Ed 2009, 4:4174–4179.CrossRef 3. Lu AH, Salabas EL, Schüth F: Magnetic nanoparticles: synthesis, protection, functionalization, and application. Angew Chem Int Ed 2007, 46:1222–1244.CrossRef 4. Tassa C, Shaw SY, Weissleder R: Dextran-coated iron oxide nanoparticles: a versatile platform for targeted molecular imaging, molecular diagnostics, and

therapy. Acc Chem Res 2011, 44:842–852.CrossRef 5. Thomas CR, Ferris DP, Lee JH, Choi E, Cho MH, Kim ES, Stoddart JF, Shin JS, Cheon J, Zink JI: Noninvasive remote-controlled release of drug molecules in vitro using magnetic actuation of mechanized nanoparticles. J Am Chem Soc 2010, 132:10623–10625.CrossRef 6. Yong KT, Roy I, Swihart MT, Prasad PN: Multifunctional nanoparticles as biocompatible targeted TPCA-1 in vitro probes for human cancer diagnosis PRKACG and therapy. J Mater Chem 2009, 19:4655–4672.CrossRef 7. Kim E, Lee K, Huh YM, Haam S: Magnetic nanocomplexes and the physiological

challenges associated with their use for cancer imaging and therapy. J Mater Chem B 2013, 1:729–739.CrossRef 8. Hui C, Shen CM, Tian JF, Bao LH, Ding H, Li C, Tian Y, Shi XZ, Gao HJ: Core-shell Fe 3 O 4 @SiO 2 nanoparticles synthesized with well-dispersed hydrophilic Fe 3 O 4 seeds. Nanoscale 2011, 3:701–705.CrossRef 9. Safi M, Courtois J, Seigneuret M, Conjeaud H, Berret JF: The effects of aggregation and protein corona on the cellular internalization of iron oxide nanoparticle. Biomaterials 2011, 32:9353–9363.CrossRef 10. Ling DS, Hyeon T: Chemical design of biocompatible iron oxide nanoparticles for medical applications. Small 2013, 9:1450–1466.CrossRef 11. Na HB, Palui G, Rosenberg JT, Ji X, Grant SC, Mattoussi H: Multidentate catechol-based polyethylene glycol oligomers provide enhanced stability and biocompatibility to iron oxide nanoparticles. ACS Nano 2012, 6:389–399.CrossRef 12. Huang CC, Tsai CY, Sheu HS, Chuang KY, Su CH, Jeng U, Cheng FY, Su CH, Lei HY, Yeh CS: Enhancing transversal relaxation for magnetite nanoparticles in MR imaging using Gd 3+ -chelated mesoporous silica shells.

The zone-center optical phonon in the zinc-blende structure is split into a doubly degenerate transverse optical (TO) mode and a longitudinal optical (LO) mode, and the Raman tensor elements are different for the TO and LO SIS3 cost modes. As calculated, the TO mode can be observed in backscattering

from the (110) and (111) surfaces, while the LO mode is allowed from the (100) and (111) surfaces [16]. In this work, we investigated single InAs NWs grown in the [111] (zinc-blende) direction. We set representing the basis of the NW crystal coordinate system. When an optical phonon is polarized along the direction , , or , its Raman tensors , , and will have only two nonzero components (d), which can be represented by a (3 × 3) matrix: (2) respectively [23]. In order to calculate the selection rules for the zinc-blende structure, the Raman tensors are transformed in two steps. First, the Raman tensors are transformed into the laboratory coordinate system with the basis . Secondly, they are rotated around the z axis by the angle θ (see Figure 1) in order to account for the additional degree of freedom of the top surface of the NWs.

this website The two transformations can be described by the matrices (3) where T denotes the transformation into the basis and S is the rotation about the NW z axis. For reasons of simplicity, we define M = ST. The Raman tensors for displacements along the directions x′ i in the basis can now be written as (4) and the Raman tensors in the basis AMP deaminase can be described by (5) Here, we have considered a backscattering

configuration along the x axis. In laboratory coordinates, the polarization of the incident radiation and the polarization of the scattered light take the form (see Figure 1) (6) depending on whether the scattered radiation is analyzed perpendicular ( ) or parallel ( ) to the wire axis, respectively. By inserting the obtained Raman tensors (Equation 5) in Equation 1, the Raman intensities of the zinc-blende structure for different configurations can be obtained. As shown in Figure 2, the theoretical intensities of the scattered light polarized perpendicular (I ⊥, polarized in the y direction) or parallel (I ∥, polarized in the z direction) to the [111] direction as a function of the angle ϕ of the incident polarization with respect to [111] are shown for TO (Figure 2a) from a bulk InAs substrate (110) in polar plots taking into account only the contribution of the Raman tensors.

Plant Soil 2003,254(2):317–327.CrossRef 65. Yang CH, Crowley DE: Rhizosphere microbial STA-9090 cell line community structure in relation to root location and plant iron nutritional status. Appl Environ Microbiol 2000,66(1):345–351.PubMedCrossRef 66. Wang Y, Ohara Y, Nakayashiki H, Tosa Y, Mayama S: Microarray analysis of the gene expression profile induced by the endophytic plant growth-promoting rhizobacteria, Pseudomonas fluorescens FPT9601-T5 in Arabidopsis. Mol Plant Microbe Interact 2005,18(5):385–396.PubMedCrossRef 67. Mathesius U, Mulders S, Gao M, Teplitski M, Caetano-Anolles G, Rolfe BG, Bauer WD: Extensive and specific responses of a eukaryote to bacterial quorum-sensing signals. Proc Natl Acad Sci U S A

2003,100(3):1444–1449.PubMedCrossRef 68. Dennis PG, Miller AJ, Hirsch PR: Are root exudates more important than other sources of rhizodeposits in structuring rhizosphere bacterial communities? FEMS Microbiol Ecol 2010, 72(3):313–327.PubMedCrossRef 69. Kuzyakov Y: Priming effects: Interactions between living and dead organic matter. Soil Biol Biochem 2010,42(9):1363–1371.CrossRef 70. Haichar FZ, Marol C, Berge O, Rangel-Castro JI, Prosser JI, Balesdent J, Heulin

T, Achouak W: Plant host habitat and root exudates shape soil bacterial community structure. ISME J 2008,2(12):1221–1230.PubMedCrossRef 71. Carvalhais LC, Dennis PG, Fedoseyenko D, Hajirezaei MR, Borriss R, von Wiren N: Root exudation of sugars, amino acids, and organic acids by maize see more as affected by nitrogen, phosphorus, potassium, and iron deficiency. Journal of Plant Nutrition and Soil Science 2011,174(1):3–11.CrossRef 72. Brune I, Becker A, Paarmann D, Albersmeier A, Kalinowski J, Puhler A, Tauch A: Under the influence of the active deodorant ingredient 4-hydroxy-3-methoxybenzyl Fenbendazole alcohol, the skin bacterium Corynebacterium jeikeium moderately responds with differential gene expression. J Biotechnol 2006,127(1):21–33.PubMedCrossRef 73. DeRisi JL, Iyer VR, Brown PO: Exploring the metabolic and genetic control of gene expression on a genomic scale. Science 1997,278(5338):680–686.PubMedCrossRef

74. Dondrup M, Albaum SP, Griebel T, Henckel K, Junemann S, Kahlke T, Kleindt CK, Kuster H, Linke B, Mertens D, et al.: EMMA 2–a MAGE-compliant system for the collaborative analysis and integration of microarray data. BMC Bioinforma 2009, 10:50.CrossRef 75. Dudoit S, Yang YH, Callow MJ, Speed TP: Statistical methods for identifying differentially expressed genes in replicated cDNA microarray experiments. Stat Sin 2002,12(1):111–139. 76. Serrania J, Vorholter FJ, Niehaus K, Puhler A, Becker A: Identification of Xanthomonas campestris pv. campestris galactose utilization genes from transcriptome data. J Biotechnol 2008,135(3):309–317.PubMedCrossRef 77. Benjamini Y, Hochberg Y: Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing.

Osteoporos Int. doi:10.​1007/​s00198-009-1052-5 2. Stöckl D, Sluss PM, Thienpont LM (2009) Specifications for trueness and precision of a reference measurement system for serum/plasma 25-hydroxyvitamin D analysis. Clin Chim Acta 408:8–13CrossRefPubMed”
“Introduction

The demonstrated efficacy of a therapy in a randomized clinical trial may not predict its actual effectiveness in clinical practice because of differences in characteristics of patients and level of medical care [1]. As a therapy for osteoporosis, the oral bisphosphonates have been widely utilized in recent years. These bisphosphonates include once-a-week alendronate (marketed in the USA since 2000), once-a-week risedronate (since 2002), and once-a-month ibandronate (since 2005). Since health data on large numbers of bisphosphonate patients buy Repotrectinib in clinical practice have now been collected (through administrative billing data, medical records, and registries), many recent observational studies have examined the effectiveness of oral bisphosphonates for reducing clinical fractures. The designs of these observational studies have included comparisons between patient populations with or without a fracture

[2, 3], with or without bisphosphonate use [4, 5], compliant or not compliant with bisphosphonate use [6–19], or between patient populations on different bisphosphonate molecules [20–23]. A key limitation in interpreting any of these comparisons is uncertainty if known or unknown differences in baseline SB525334 fracture risk between patient populations could account for some or all of the reported results. An approach to directly measure the baseline risk of an outcome within patient populations that has been used in effectiveness studies of other therapies may be applicable to the study of bisphosphonates. In a comparison of patients receiving a bare or drug-eluting stent,

the mortality 2 days after procedure was G protein-coupled receptor kinase used to assess risk of the mortality outcome independent of possible drug effect [24]. In a comparison of patients receiving influenza vaccine or not, the mortality after vaccination but before flu season was used to assess risk of mortality outcome independent of possible vaccination effect [25]. Likewise, following initiation of bisphosphonate therapy, the realization of fracture reduction is likely not immediate. Bone mineral density, a surrogate marker of therapeutic effect, begins to change after start of therapy though does not reach its maximum level of change until at least 1 year on therapy [26]. As changes in bone density and quality take time, correspondingly, fracture reductions have not been noted earlier than 6 months after start of therapy within post hoc, pooled analysis of clinical trials [27, 28].

I fondly remember Govindjee’s

effort to learn Chinese phrases. The first phrase he wanted to learn was “You la you ma?” (Is there any chilli sauce?), so that he could spice up his food. Since that meeting in China, Govindjee and I have been in yearly contact, usually around Christmas time. Indeed he came to Canberra for a sabbatical in 1998, and we joined forces with other colleagues to publish a paper in the Indian Journal of Biochemistry and Biophysics, as he insisted (Chow et al. 2000). Roberta Croce Professor of Biophysics and Photosynthesis University of Amsterdam, Amsterdam, The Netherlands I am looking at my first picture with Govindjee; it was quite some time ago but it seems that find more I am the only one that is getting older here. I was very proud to get a picture with the famous Govindjee but I am even more happy to have many more pictures with him now, which

means that I have enjoyed his company and been overwhelmed by his enthusiasm for science many times. On top of his scientific achievements, Govindjee is a very strong catalyzer for the photosynthetic community, the guardian of our history and a great example of scientific passion. I am looking forward to our next picture! Henry Daniel Professor, Penn Dental Medicine University of Pennsylvania, EPZ-6438 cell line Philadelphia, PA Govindjee’s visit to Madurai many years ago changed my life and career! Because he dazzled us with his writing/editorial/presentation and scientific accomplishments, I wanted to follow his footsteps and so joined the University of Illinois at Urbana-Champaign, declining other offers from UK and Europe. Even though I didn’t pursue research in photosynthesis, I am still using the chloroplast system for various Cobimetinib biomedical applications. In addition, I am now the Editor-in-Chief of

the Plant Biotechnology Journal (ranked in the top 5 among 200 plant science journals), again following Govindjee’s footsteps. So, I thank him for his leadership that profoundly influenced my life and career. Mrinmoyee Das Retired Professor of Chemistry Kolkota, India Govindjee, as I know him I remember vividly the picture of my first meeting with Govindjee on the 14th of October 1965, around 10 pm, when my flight from Chicago landed at the Champaign airport. I had come from Kolkata, India, to join the research group of Eugene Rabinowitch at the University of Illinois at Urbana-Champaign; I was to be a post-doctoral research associate. The flight from London to Chicago was delayed by almost 7 h due to bad weather in London, and I was on the last flight for that day from Chicago to Champaign.

The PPy nanotube diameter can be enhanced by forming thicker

ZnO nanorod array core structure. However, this reduces the effective thickness of PPy tubular sheath and hence the effective mass of PPy which is an active component for charge storage. On the other hand, increasing thickness of PPy by electropolymerization for longer pulsed current cycles excessively covers the top of the ZnO nanorod arrays making it difficult to etch away the ZnO core which prevented realization of PPy nanotubular arrays. Figure 3 shows the ZnO core-PPy sheath structure with the thicker PPy layer deposited using 20 k unipolar pulsed current cycles. This results in formation of thick conjoined PPy sheath with thickly deposited PPy over the top of ZnO nanorods (Figure 3A). Figure 3B shows a cross-sectional view indicating the ZnO nanorods could still be coated with PPy along its length. The side panel Selleck Screening Library in Figure 3C shows conjoined PPy sheath over ZnO nanorods of average diameter approximately 985 nm to 1 μm. Morphology of the thick PPy deposit is like nodules. Figure 3D shows the top view of the PPy coated ZnO nanorods tips. Figure 3E shows the same view after ammonia etching for 4 h. It is evident that such ZnO nanorod core-PPy sheath

structure did not result in the PPy nanotube selleckchem structure after etching. The evolution of the PPy sheath and nanotube structure is schematically shown in Figures 4A, B, C, D, E, F. The vertical ZnO nanorod array (Figure 4A) is preferentially coated with PPy by pulsed

electropolymerization process through surfactant action. Progressively, on continued pulsed current polymerization cycles, the PPy sheath thickness increases (Figure 4B) with possible merging of PPy sheath walls (Figure 4C). Figures 4D, E, F show the evolution of PPy nanotubes through etching of ZnO core starting at the nanorod tips which after short term etching results in the PPy nanotubes along with the inverted conical ZnO cladding (Figure 4D). The PPy nanotube arrays without the ZnO cladding are created by complete etching Rho of ZnO for longer periods as depicted in Figure 4E with an open pore structure as shown in the top view in Figure 4F. Figure 2 SEM images of ZnO nanorod arrays coated with pulsed current polymerized PPy sheath. (A) Initial stage of PPy oligomers cluster deposition, (B) ZnO core-PPy sheath structure after 10 k pulsed electropolymerization cycles, (C) PPy nanotube array after 2-h etch, and (D) open pore PPy nanotube array after 4-h etch. Figure 3 SEM images. (A) Thicker PPy deposited over ZnO nanorod array when electropolymerization was carried out for 20 k pulsed current cycles, (B) cross-sectional view of PPy sheath coated along the ZnO rod length, and (C) conjoined view of PPy sheath over ZnO nanorods with average diameter of 985 nm. Top view of ZnO nanorod tips with thick PPy sheath (D) before etch and (E) after ammonia etch.

Mutation of this gene produces a non-toxigenic phenotype relative to the wt check details strain. However, the relationship of desI with phaseolotoxin synthesis is still unknown [12]. Additionally, it has been observed that mutation in the desI gene decreases the growth rate at 18°C relative to the wt strain, suggesting a cold-sensitivity in the mutant strain (unpublished data). Another of the mechanisms reported to be involved in membrane lipid composition changes correspond to de novo synthesis. The fabF and lpxP genes induced by low temperature participate in this process [33]. β-ketoacyl-ACP synthase II, the fabF gene product, converts palmitoleic acid to cis-vaccenic acid, which is in turn transferred by an acyltransferase

(LpxP) into lipid A, a component of polysaccharides [33, 34]. Although these two genes were not found in our microarray, several genes involved in cell wall biogenesis and membrane synthesis were identified (Cluster 4). These include the murA gene (PSPPH_4139) that is involved in peptidoglycan synthesis (a major component of cell wall), the PSPPH_4682 gene involved in polysaccharide synthesis, as well as three genes PSPPH_4669, PSPPH_3226, see more and galU (PSPPH_2260) that encode an acetyl-, glycosyl- and uridyl- transferase, respectively, which are likely associated with the transfer of these groups during polysaccharides synthesis.

Additionally, it has been demonstrated that during cell envelope biogenesis, there is an increase in outer membrane lipoproteins, which increase connections with the cell wall [34, 35]. In our analyses four genes (PSPPH_ 1464, PSPPH_2654, PSPPH_2842, and PSPPH_3810) encoding lipoproteins were induced, which may be related to outer membrane synthesis. The microarray results suggest that membrane component synthesis is activated in the conditions of our study and these changes are likely related to cell envelope remodeling to adapt to low temperatures. Low temperature induces expression of motility genes in P. syringae pv. phaseolicola NPS3121 Cluster 5 comprises genes induced at 18°C that

are involved in bacterium motility. The data suggest that chemotaxis and rotation of flagella processes function in low temperatures on P. syringae pv. phaseolicola NPS3121. Two genes, PSPPH_3880 that encodes the membrane-bound methyl accepting chemotaxis Thymidine kinase protein (MCP)-like receptor WspA, and PSPPH_3881, that encodes the CheW-like scaffolding protein WspB, showed high transcripts levels at 18°C relative 28°C (Table 1). WspA and WspB are related to the chemotaxis process. Chemotaxis, as well as other types of taxis (e.g., thermotaxis), enables bacteria to approach beneficial environments and escape from hostile ones. Depending on the parameter monitored, bacteria will respond by either swimming toward attractants or retreating from repellants. Thus, the signal sensed by chemotaxis causes changes in flagellum motility [36].

In cases of uncertain preoperative diagnosis in septic and unstable patients, laparoscopy can shorten the observation period and avoid the need for imaging test [27]. Source control Source control encompasses all measures undertaken to eliminate the source of infection and to control ongoing contamination. The most common source of infection in community acquired

intra-abdominal infections is the appendix, followed by the colon, and then the stomach. Dehiscences complicate 5-10% of intra-abdominal bowel anastomoses, and are associated with a mortality increase [3]. Timing and adequacy of source control are the most important issues in the management of intra-abdominal infections, because inadequate and late operation may have a negative effect on the outcome. Early control of the septic source can be achieved either by nonoperative or operative means. Nonoperative interventional check details procedures include percutaneous drainages of abscesses. Ultrasound and CT guided percutaneous drainage of abdominal and extraperitoneal abscesses in selected patients are safe and effective. Numerous studies in the surgery and radiology literature have documented the effectiveness of percutaneous drainage in selected patients, with cure rates of 62%-91% and with

morbidity and mortality rates equivalent to FK228 ic50 those of surgical drainage [32–39]. The principal cause for failure of percutaneous drainage is misdiagnosis of the magnitude, extent, complexity, location of the abscess [40]. Surgery is the most important therapeutic measure to control intra-abdominal infections. Generally, the choice of the procedure depends on the anatomical source of infection, on the degree of peritoneal inflammation, on the generalized septic response and on the patient’s general conditions. Surgical source control entails resection or suture of a diseased or perforated viscus

(e.g. diverticular perforation, gastroduodenal perforation), removal of the infected organ (e.g. appendix, gall bladder), debridement of necrotic tissue, resection of ischemic see more bowel and repair/resection of traumatic perforations. Laparotomy is usually performed through a midline incision. The objectives are both to establish the cause of peritonitis and to control the origin of sepsis. Appendicitis Acute appendicitis is the most common intra-abdominal condition requiring emergency surgery. Acute appendicitis is the most common intra-abdominal condition requiring emergency surgery. Studies have demonstrated that antibiotics alone may be useful to treat patients with early, non perforated appendicitis, even if there is a risk of recurrence [41]. In 1995, Eriksson and Granstrom [42] published the results of a randomized trial of antibiotics versus surgery in the treatment of appendicitis. All patients treated conservatively were discharged within 2 days, except one who required surgery because of peritonitis secondary to perforated appendicitis.

Conclusions Our comparative XPS, TDS, and selleck chemicals llc AFM studies of Ag-covered L-CVD SnO2 nanolayers deposited on atomically clean Si(111) substrate and subsequently exposed to air showed the following: As deposited L-CVD SnO2 nanolayers (20-nm thickness) covered with 1 ML of Ag consisted a mixture of tin oxide SnO and tin dioxide SnO2 with the relative [O]/[Sn] concentration of approximately 1.3. After long-term dry air

exposure of the Ag-covered L-CVD SnO2 nanolayers, they were still a mixture of tin oxide (SnO) and tin dioxide (SnO2) phases with slightly increased [O]/[Sn] ratio of approximately 1.55, related to the adsorption of oxygen containing residual air gases from the air; moreover, an evident increase of C contamination was observed with [C]/[Sn] ratio at approximately 3.5, whereas surface Ag atoms concentration was twice smaller. After registration of TDS spectra, the non-stoichiometry of Ag-covered L-CVD SnO2 nanolayers goes back to 1.3, whereas C contamination evidently decreases (by factor of 3)

but cannot be completely removed in this process. Simultaneously, Ag Selonsertib concentration subsequently decreased by factor of approximately 2, which was related to the diffusion of Ag atoms into the subsurface layers related to the grain-type surface/subsurface morphology, as confirmed by XPS ion depth profiling studies. The variation of surface chemistry of Ag-covered L-CVD SnO2 nanolayers before and after registration of TDS spectra observed by XPS was

in a good correlation with the desorption of residual gases like H2, H2O, O2, and CO2 from these nanolayers observed in TDS experiments. All the observed experimental facts testified the limited sensing application of L-CVD SnO2 nanolayers, corresponding to the long response/recovery times, for instance, in NO2 atmosphere, as was observed some years ago by group of Larciprete [13]. However, their electronic and sensing properties are still currently under investigation in our group. Acknowledgements This work was realized within the Statutory Flavopiridol (Alvocidib) Funding of Institute of Electronics, Silesian University of Technology, Gliwice, and partially financed within the Operation Program of Innovative Economy project InTechFun: POIG.01.03.01-00-159/08. References 1. Göpel W, Schierbaum K-D: SnO 2 sensor: current status and future progress. Sensors Actuators 1995, B26–27:1–12.CrossRef 2. Comini E, Faglia G, Sberveglieri G (Eds): Electrical based gas sensors In Solid State Gas Sensing. New York: Springer; 2009:47–108. 3. Carpenter MA, Mathur S, Kolmakov A: Metal Oxide Nanomaterials for Chemical Sensors. New York: Springer; 2013.CrossRef 4. Lantto V, Mizsei J: H 2 S monitoring as an air pollutant with silver-doped SnO 2 thin-film sensors. Sensors Actuators 1991, B5:21–25.CrossRef 5.

TNF and IL-12 assays For the TNF secretion assays, 2

× 105 bone marrow-derived macrophages in DMEM infection media were seeded onto each well of 12 well plates and infected with bacteria as indicated above. The culture supernatants were then collected 20 h after incubation in infection media supplemented with 100 μg/ml gentamycin. The amount of TNF in supernatants was then measured via ELISA (BD Bioscience). The RAW IL-12 promoter cell line was created and used to measure IL-12 p40 induction as described in great detail in our previous publication [12]. TLR interaction assay The Chinese hamster ovary (CHO) cells transfected with the inducible membrane protein CD25 under control of a region from the human E-selectin promoter containing nuclear fact-kB Selleck MM-102 binding sites and expressing CD14 and either human Toll-like receptor 2 (TLR-2) or human TLF-4 were created as described in [28] kindly provided by Dr. D.T. Golenbock. Cells were used exactly as described previously by our group [12]. Apoptosis assays In most of the experiments the flow cytometry-based, hypodiploid assay was used for the detection of apoptosis after infection of bone marrow-derived macrophages and dendritic Epacadostat in vivo cells. Cells were collected

after infection, pelleted and resuspended in propidium iodine (PI)/RNase buffer (BD Pharmingen) for 20 min and the percentage of hypodiploid positive cells was determined by flow cytometry in duplicates in the FL-2 channel at 580 nm (FACS-Calibur, BD Biosciences). The TUNEL assay was performed as suggested by the manufacturer (Roche) and described previously [8]. The apoptosis induction mediated by lipoglycanes was analyzed via AnnexinV-Alex488 (Molecular Probes) and PI double staining and flow cytometry as described previously [12]. Caspase inhibition and TNF neutralization assays BMDMs from BALB/c mice were treated with a pan-caspase-3/6/7 inhibitor (100 μM), caspase-3 inhibitor negative control (100 μM) (both from Calbiochem), anti murine TNF neutralizing antibody (5 μg/ml), isotype control antibody (5 μg/ml) (both from BD Bioscience), or pentoxifylline (Sigma, 100 μg/ml) for 1 h at 37°C/5% CO2 then infected with Meloxicam M. smegmatis at MOI 10:1 for 2 h as described above. Cells

were then washed 3 times in PBS and incubated for an additional 20 h in DMEM infection media supplemented with the appropriate inhibitors and controls mentioned above and the apoptosis assay was performed. ROS detection assay Reactive oxygen species in BMDM and BMDD cells were detected at 2 h post infection using the ROS sensitive dye dihydroethidium (DHE) (Invitrogen). BMDM or BMDD cells were deprived of L929 supernatant or rGM-CSF respectively 16 hrs prior to infection and maintained in cytokine free media without phenol red for the length of the experiment. Post infection, cells were washed once in HBSS and then incubated in 2 μM DHE for 15 minutes. Cells were washed 3 times with HBSS, fixed with 4% paraformaldehyde and analyzed by flow cytometry.