They bind to DNA [3, 5] preferring AT-rich DNA-sequences [11] as

They bind to DNA [3, 5] preferring AT-rich DNA-sequences [11] as well as to laminin, hyaluronic acid, heparin, and chondroitin sulphate [5, 6, 12]. The data available so far portray Hlp as multi-faceted proteins, and accordingly a NVP-HSP990 variety of possible functions have been ascribed to Hlp. Hlp were suggested to impact DNA packaging, protection of DNA from enzymatic and non-enzymatic strand breakage [11], gene regulation [1], nucleic acid metabolism, non-homologous-end-joining repair [13], adaptation to Thiazovivin hypoxic conditions [2],

induction of dormancy [2], adaptation to cold shock [14], adhesion [6, 9, 12, 15–17], cell wall biogenesis [10] and regulation of growth rate [1, 5, 10]. A role in transition to the non-culturable state and in resuscitation from the non-culturable state was shown in M. smegmatis[18]. Whiteford et al. [19] investigated the growth characteristics of an M. smegmatis with a deletion of hlp. They found that the mutant showed less aggregation in broth cultures. Furthermore, they observed an increased sensitivity towards Isoniazid. The M. smegmatis mutant also was affected in UV-resistance and resistance towards freezing/thawing. Takatsuka et al. [20] have recently shown that Hlp has a similar activity to ferritin superfamily proteins and protects DNA by ferroxidase activity. It furthermore captures iron molecules and functions selleck as iron storage protein. Approaches to elucidate the

functions of Hlp by mutagenesis did not always confirm the expected roles of Hlp [2, 15, 21]. Our own attempts to generate a MDP1 deletion mutant had failed. Furthermore and in line with our own experience, Sassetti et al. [22] had shown by high density mutagenesis that the gene Rv2986c from M. tuberculosis, which is homologous to MDP1 from BCG, is required for optimal growth of M. tuberculosis. We therefore followed the strategy BCKDHB to analyse Hlp functions by down-regulation of Hlp expression by antisense-technique. Advantages of this technique are the possibility to analyse essential genes and to repress genes present in several copies. In mycobacteria the antisense-technique

has been applied to down-regulate ahpC from M. bovis[23], dnaA from M. smegmatis[24], FAP-P from M. avium subsp. paratuberculosis[25] or pknF from M. tuberculosis[26]. In a previous study we described the generation of the antisense-strain M. bovis BCG (pAS-MDP1) which carries the plasmid pAS-MDP1 causing a reduction of MDP1 expression in BCG by about 50% [27]. We analysed BCG (pAS-MDP1) with respect to general growth characteristics. The down-regulated BCG grew faster in broth culture and achieved a higher cell mass in the stationary phase. Similarly, growth was enhanced in human and murine macrophage-like cell lines. A further important finding was the reduced protein synthesis occurring under hypoxic conditions [27]. These findings support a role of MDP1 in growth regulation of M. bovis BCG.

Only identities above 90% are shown (PDF 25 KB) References 1 Co

Only identities above 90% are shown. (PDF 25 KB) References 1. Cox ML: Red palm weevil, Rhynchophorus ferrugineus in Egypt. FAO Pl Prot Bul 1993, 41:30–31. 2. Butera G, Ferraro C, Colazza S, Alonzo G, Quatrini P: The selleck screening library culturable bacterial community of frass produced by larvae of Rhynchophorus

ferrugineus Olivier (Coleoptera: Curculionidae) in the Canary island date palm. Lett Appl Microbiol 2012, 54:530–536.PubMedCrossRef 3. RGFP966 mouse Dembilio Ó, Jacas JA: Basic bio-ecological parameters of the invasive Red Palm Weevil, Rhynchophorus ferrugineus (Coleoptera: Curculionidae), in Phoenix canariensis under Mediterranean climate. Bull Entomol Res 2011, 101:153–163.PubMedCrossRef 4. Faleiro JR: A review on the issues and management of red palm weevil Rhynchophorus ferrugineus (Coleoptera: Rhynchophoridae) in coconut and date palm during the last one hundred years. Internat J Trop Insect Sci 2006, 26:135–154. 5. Ferry M, Gomez S: The Red Palm Weevil in the Mediterranean area. Palms 2002, 46:172–178. 6. Engel P, Moran NA: The gut microbiota of insects – diversity in structure and function. FEMS Microbiol Rev 2013, 37:699–735.PubMedCrossRef 7. Hongoh Y: Diversity and genomes

of uncultured microbial symbionts in the termite gut. Biosci Biotechnol Biochem 2010, 74:1145–1151.PubMedCrossRef selleck chemical 8. Colman DR, Toolson EC, Takacs-Vesbach CD: Do diet and taxonomy influence insect gut bacterial communities? Mol Ecol 2012, 21:5124–5137.PubMedCrossRef 9. Rosenberg E, Zilber-Rosenberg I: Symbiosis and development: the hologenome concept. Birth Defects Res (Part C) 2011, 93:56–66.CrossRef 10. Dillon RJ, Dillon VM: The gut bacteria of insects: non-pathogenic interactions. Annu Rev Entomol 2004, 49:71–92.PubMedCrossRef 11. Kaufman MG, Klug MJ: The contribution of hindgut bacteria to dietary carbohydrate utilization by crickets (Orthoptera, Gryllidae). Comp Biochem Physiol A Physiol 1991, 98:117–123.CrossRef 12. Chakravorty S, Helb D, Burday M, Connell N, Alland D: A detailed analysis of 16S ribosomal RNA gene segments

for the diagnosis of pathogenic bacteria. Cisplatin concentration J Microbiol Methods 2007, 69:330–339.PubMedCentralPubMedCrossRef 13. Tang X, Freitak D, Vogel H, Ping L, Shao Y, Cordero EA, Andersen G, Westermann M, Heckel DG, Boland W: Complexity and variability of gut commensal microbiota in polyphagous lepidopteran larvae. PLoS One 2012,7(7):e36978. doi:10.1371/journal.pone.0036978CrossRefPubMedCentralPubMed 14. Shivaji S, Chaturvedi P, Suresh K, Reddy GS, Dutt CB, Wainwright M, Narlikar JV, Bhargava PM: Bacillus aerius sp. nov., Bacillus aerophilus sp. nov., Bacillus stratosphericus sp. nov. and Bacillus altitudinis sp. nov., isolated from cryogenic tubes used for collecting air samples from high altitudes. Int J Syst Evol Microbiol 2006, 56:1465–1473.PubMedCrossRef 15. Suzuki T, Yamasato K: Phylogeny of spore-forming lactic acid bacteria based on 16S rRNA gene sequences. FEMS Microbiol Lett 1994, 115:13–17.PubMedCrossRef 16.

We also performed ROC curve analysis for the three significant ge

We also performed ROC curve analysis for the three significant genes, singly or in combination, considered as continuous variables. Resultant AUCs were 0.5917 for HIC1, 0.6725 for RASSF1 and 0.5409 for GSTP1, the best AUC (0.6959) reached for the combination of the three genes (Figure 4). Figure 4 ROC curves relating to the three significant genes (HIC1, RASSF1, GSTP1) analyzed Vorinostat in vitro singly or in combination. Recurrence-free survival analysis of patients with methylated or unmethylated tumors highlighted a significantly higher recurrence-free survival (P = 0.0019) for those whose tumors showed the methylated phenotype (Figure 5). Figure 5 Recurrence-free survival in patients

with methylated phenotype (samples with at least one of the three significant genes methylated) or unmethylated phenotype (samples with none of the three genes methylated) . The recurrence free survival analysis performed considering only the recurrent patients, showed that patients with unmethylated tumors had a lower median recurrent free survival time (14.5 months), with the respect to patients with methylated ones (18 months). However, the two subgroups are not equal distributed to give a statistical significant result (P = 0.9392, data not shown). Multivariable analysis considering clinical and biological parameters (patient age and sex; tumor grade, stage and size; tumor multiplicity, methylated phenotype) showed that only age and

methylated phenotype were independent predictors Z-DEVD-FMK nmr of recurrence. Specifically, patients under 70 years of age showed a higher probability of relapsing than older ones (P = 0.028) and their methylation phenotype was significantly predictive of recurrence (P < 0.0001). Discussion The present study focused on evaluating the methylation status of tumor suppressor genes and on verifying its role in predicting recurrence

of non muscle invasive bladder cancer (NMIBC). The MS-MLPA technique has the advantage of requiring only a small quantity of DNA, is capable of rapidly determining the methylation status of numerous genes in the same experiment, and has also been shown to work well in formalin-fixed paraffin-embedded samples. However, an important limitation of our study was the lack of a sufficient Oxymatrine quantity of cancer tissue to confirm the methylation results using a second technique such as methylation specific PCR (MS PCR) or gene expression analyses. In agreement with results from other studies [18], we found a positive correlation between gene methylation and lack of recurrence, highlighting that putative tumor suppressor genes do not always act as tumor suppressors but may actually have different biological functions. Statistical analysis revealed 3 genes (HIC1, GSTP1, and RASSF1) capable of significantly predicting tumor recurrence. Their methylation was significantly indicative of a lack of recurrence at the 5-year follow up.

burnetii infected THP-1 cells regardless of ongoing bacterial pro

burnetii infected THP-1 cells regardless of ongoing bacterial protein synthesis. These results confirm that genes with significant mRNA expression changes by oligonucleotide microarrays analysis are differentially expressed when measured by RT-qPCR. Figure 4 selleck chemicals RT-qPCR of selected genes confirms microarray expression trends. A, shows the microarray data of the BI 10773 molecular weight genes used to confirm microarray expression trends. Fold difference (-CAM)

is the fold change of differentially expressed THP-1 genes in response to C. burnetii infection after mock treatment. Fold difference (+CAM) is the fold change of differentially expressed THP-1 genes in response to C. burnetii infection after CAM treatment. B, difference in mRNA levels in selected genes relative to β-actin. An equal amount of total RNA from each sample was analyzed by RT-qPCR. The Y-axis represents fold changes

in gene expression while X axis shows the conditions under which gene expression was observed (mock and CAM treated, and uninfected and C. burnetii infected THP-1 cells). U-CAM, uninfected THP-1 minus CAM. U+CAM, uninfected THP-1 plus CAM. I-CAM, infected THP-1 minus CAM. I+CAM, infected THP-1 plus CAM. The results represent the mean of three biological samples and three technical replicates of each sample. Error bars represent the s.e.m. Discussion Bacterial effector proteins are crucial to the survival and growth of intracellular pathogens within the eukaryotic cellular environment. These interactions may be at a myriad of pathways or Inhibitor Library screening at points within a single pathway. Moreover, the growth of C. burnetii within the lumen of the PV would require the mediation of interactions with the host cell using effector proteins, which are predicted to be delivered by the pathogen’s type IV secretion system [10, 11, 19]. The goal of this study was to identify host genes that are specifically manipulated by C. burnetii proteins. Our hypothesis was that the Calpain expression of host cell genes will be changed by infection with C. burnetii NMII and that the expression of a subset of these genes will be directly affected by ongoing

bacterial protein synthesis. Identification of such genes will aid in the understanding of host molecular mechanisms being targeted by C. burnetii during growth. In order to identify the host genes regulated by C. burnetii proteins, we compared CAM and mock treated mRNA profiles of THP-1 cells following a 72 h infection with C. burnetii. Microarray data analysis shows that the majority of host genes were up- or down regulated similarly in both the mock and CAM treated array sets, suggesting that most THP-1 genes were not differentially modulated at the RNA level by active C. burnetii protein synthesis. We had predicted that the majority of expression changes in the host cell would be in response to the physical presence of bacteria within the cell.

I coefficient is the product I = C c   × C E   × C M   × C R , wh

I coefficient is the product I = C c   × C E   × C M   × C R , where C c is the cross-correlation coefficient pertaining to whole thermograms, termed “p-t curves”. Other

factors, termed “specific coefficients”, pertain to different parameters of the thermogram: E is “a measurement of the total energy dissipated by the culture during its growth”; M is “the maximum value of the dissipated power”; R, “the maximum metabolic rate”, is the maximum value of the time-derivative of the heat flow. Selleck AZD8186 The initial approach [26] was further developed [27] with the inclusion of the thermogram time-derivative, called “t-d curve” into a more complex “discriminant analysis” that was able to objectively evidence differences between strain growth patterns. One may easily notice the equivalence of some of the above parameters with quantities utilized in the present paper: E ↔ ΔH tot and M ↔ HF max , respectively. There is another natural similarity between the two approaches which involves the well-defined growth conditions, a normal requirement for comparing the growth of different cultures. Besides the differences in statistical/mathematical processing, one may outline several differences between the two methods. One may use the term “overall” for the method of Bermúdez, López et al., with a double-meaning: (i)

the whole growth thermogram is needed for all key quantities C c , C E , C M , C R ; (ii) the raw thermal signal, consisting of several overlapping metabolic processes is subject to statistical analysis. In fact, the authors seek for maximum complexity of growth (by adjusting the culture medium) as a necessary selleck chemicals llc condition for discrimination between species. The present study involves both “overall” and “local” aspects: (i)’ the whole thermogram is mafosfamide needed for decomposition and ΔH tot evaluation; (ii)’ discrimination parameters are looked for in component (local) features of the thermogram, with some (possible) metabolic significance. The present study may be regarded as a start for further, extended investigations for other species and strains. Optimization

of the advanced procedure for different thermal data is straightforward. As obtaining of sufficient data is time-consuming with single-channel microcalorimeters, the presented analysis was intended to avoid Lamprecht’s [28] caveat: “In our high-tech time of stream-lined instruments with black-box character, we experience automatic inputs, outputs, and computer calculations that do not allow getting to the roots of the thermal data”. Conclusions Bacterial populations of Staphylococcus aureus and Escherichia coli exhibit different microcalorimetric growth patterns in both qualitative and check details quantitative assessments. The devised experimental routine (based on thermograms obtained from samples kept in cold storage, sealed in the measuring batch cells [7]) is sufficiently reproducible and accurate.

Figure 2 Schematic diagram showing the process of fabricating the

Figure 2 Schematic diagram showing the process of fabricating the sTNP tip. (a, b) Etching process for reflective metal layer on Olympus RC-800 Si3N4 tip. (c) The vertex of the tip was flattened by scanning the tip across a polished Si3N4 wafer. (d, e) Present SEM images of the Si3N4 AFM tip before and after the scanning process, respectively. (f) A small quantity of adhesive was applied to the flat top of the AFM tip. (g) Attached sTNP to the vertex of the flattened tip with adhesive followed

by curing. (h) Schematic diagram of fabricated sTNP tip. (i, j) SEM images of the sTNP tip. The experimental setup of the deposition of OSI-906 charge to the sTNP tip The experimental setup used for the deposition of charge to the sTNP tip is presented in Figure 3. The back side of the sTNP tip was affixed to the 30-nm Au/ 20-nm Ti-coated glass slide using conductive copper tape (3 M, St. Paul, MN, USA). A 50-nm Ti-coated tipless cantilever (CSC12, MikroMasch, Tallinn, Estonia) was mounted on the JPK AFM scanner as the top electrode. The end of the tipless cantilever was positioned precisely on the sTNP at the vertex of the Si3N4 tip by aligning the JPK AFM scanner under an inverted optical microscope (IX 71, Olympus; Figure 3b). DC voltage (−2.5 kV) was applied to the tipless cantilever for 90 s under air, and the 30-nm Au/20-nm Ti-coated glass slide was used as the

ground for the deposition of the negative GS-1101 manufacturer charge to the sTNP tip. The force-distance (f-d) curves of the

sTNP tip on the grounded gold surface were used to verify whether the charge was deposited [17]. Figure 3 Schematic diagram of experimental setup for the deposition of charge to the sTNP tip. (a) Schematic diagram of experimental setup for the deposition of charge to the sTNP at the vertex of the Si3N4 AFM tip and (b) × 40 optical microscope image of the charging setup. Measurement of the electrostatic fields The charged sTNP tip was then used for the measurement of f-d curves to determine the electrostatic field beside the top electrode of the parallel plate condenser (Figure 1). The sTNP tip is located slightly inward at the end of the AFM cantilever; therefore, the end of PAK5 the AFM cantilever is susceptible to striking the edge of the top electrode when the distance between the AFM tip and the electrode is within 10 μm. To overcome this situation, 21 spots spaced at 0.25 μm along the X-axis at a distance of 10 to 15 μm are selected for the measurement of the f-d curves in order to derive the electrostatic field. As shown in Figure 1, the edge center of the condenser was plotted as the origin of the X- and Z-axes. DC voltage (V app) of ±25 V was applied on the top electrode, and the bottom electrode was left grounded. Each curve measurement was conducted for distances of 15 μm along the Z-axis, from 6 μm below to 9 μm above the top electrode. The ramp rate and the ramp size of each f-d curve were 2 Hz and 15 μm, respectively.

CXCR3 has now been identified in many cancers including osteosarc

CXCR3 has now been identified in many cancers including osteosarcoma and CXCR3 ligands were expressed by lungs which selleck inhibitor are the primary sites to which this tumor metastasize. This study tested the hypothesis that disruption of the CXCR3/CXCR3 ligands complexes could lead to a decrease in lungs metastasis. The experimental design involved the use of the CXCR3 antagonist, AMG487, and two murine models of osteosarcoma lung metastases.

Following tail vein injection of osteosarcoma cells, mice that were systematically treated with AMG487 according to preventive or curative protocols had a significant reduction in metastatic disease. Treatment of osteosarcoma cells in vitro with AMG487 led to decreased migration, decreased matrix metalloproteinase activity, decreased proliferation/survival and increased caspase-independent death. Taken together, our results support the hypothesis that CXCR3 and their ligands intervene in the initial dissemination of the osteosarcoma cells to the lungs and stimulate the growth and expansion of the metastatic foci in later stages. Moreover, these studies indicate that targeting CXCR3 may specifically inhibit tumor Selleckchem MI-503 metastasis without adversely affecting antitumoral

host response. Poster No. 200 Systems Biology: A Therapeutic Target for Tumor Therapy Albrecht Reichle 1 , Thomas Vogt1 1 Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany Tumor-related activities that seem to be operationally Akt inhibitor induced by the division of function, such as inflammation, neoangiogenesis, Warburg effect, immune response, extracellular matrix remodeling, cell proliferation rate, apoptosis, coagulation effects, present itself from a systems perspective

as an enhancement of complexity. We hypothesized, that tumor systems-directed therapies might have the capability to use aggregated action effects, as adjustable sizes to therapeutically modulate the tumor systems’ stability, homeostasis, and robustness. We performed a retrospective analysis of recently published data on 266 patients with advanced and heavily pre-treated (10% to 63%) vascular sarcoma, melanoma, renal clear cell, cholangiocellular, and hepatocellular carcinoma, hormone-refractory prostate cancer, gastric cancer, and multivisceral Langerhans’ Cediranib (AZD2171) cell histiocytosis enrolled in ten multi-center phase II trials (11 centers). Each patient received a multi-targeted systems-directed therapy that consisted of metronomic low-dose chemotherapy, a COX-2 inhibitor, combined with one or two transcription modulators, pioglitazone +/− dexamethason or IFN-alpha. These treatment schedules may attenuate the metastatic potential, tumor-associated inflammation, may exert site-specific activities, and induce long-term disease stabilization followed by prolonged objective response (3% to 48%) despite poor monoactivity of the respective drugs. Progression-free survival data are comparable with those of reductionist-designed standard first-line therapies.

Mol Syst Biol 2007, 3:91 PubMedCrossRef 8 Kohanski


Mol Syst Biol 2007, 3:91.PubMedCrossRef 8. Kohanski

MA, Dwyer DJ, Hayete B, Lawrence CA, Collins JJ: A common mechanism of cellular death induced by bactericidal antibiotics. Cell 2007,130(5):797–810.PubMedCrossRef 9. Wang X, Zhao X: Contribution of oxidative damage to antimicrobial lethality. Antimicrob Agents Chemother 2009,53(4):1395–1402.PubMedCrossRef 10. Cheng B, Annamalai T, Sorokin E, Abrenica M, Aedo S, Tse-Dinh YC: Asp-to-Asn substitution at the first position of the DxD TOPRIM motif of recombinant bacterial topoisomerase I is extremely lethal BIBW2992 to E. coli . J Mol Biol 2009,385(2):558–567.PubMedCrossRef 11. Cheng B, Shukla S, Vasunilashorn S, Mukhopadhyay S, Tse-Dinh YC: Bacterial cell killing mediated by topoisomerase I DNA cleavage activity. J Biol Chem 2005,280(46):38489–38495.PubMedCrossRef 12. Sutherland JH, Cheng B, Liu IF, Tse-Dinh YC: SOS induction by stabilized topoisomerase IA cleavage complex occurs via the RecBCD

pathway. J Bacteriol 2008,190(9):3399–3403.PubMedCrossRef 13. Liu IF, Annamalai T, Sutherland JH, Tse-Dinh YC: Hydroxyl radicals are involved in cell killing by the bacterial topoisomerase I cleavage complex. J Bacteriol 2009,191(16):5315–5319.PubMedCrossRef 14. Partridge JD, Scott C, Tang Y, Poole RK, Green J: Escherichia coli Transcriptome Dynamics during the Transition from Anaerobic to Aerobic Conditions. J Biol Chem 2006,281(38):11230–11237.CrossRef 15. Partridge JD, Sanguinetti G, Dibden DP, Roberts RE, Poole RK, Green J: Transition of Escherichia coli from Aerobic to Micro-aerobic Conditions Involves Fast and Slow Reacting Regulatory Components. J Biol Chem 2007,282(15):11230–11237.PubMedCrossRef 16. Kang Y, Weber KD, Qiu Y, Kiley PJ, Blattner FR: Genome-wide expression analysis indicates that FNR of Escherichia coli K-12 regulates a large number of genes of unknown function. J Bacteriol 2005,187(3):1135–1160.PubMedCrossRef 17. Salmon K, Hung SP, Mekjian Resminostat K, Baldi P, Hatfield GW, Gunsalus RP: Global gene expression profiling in Escherichia coli K12. The effects of oxygen availability and FNR. J Biol Chem 2003,278(32):29837–29855.PubMedCrossRef 18. Scott C, Partridge JD,

Stephenson JR, Green J: DNA target sequence and FNR-dependent gene expression. FEBS Lett 2003,541(1–3):97–101.PubMedCrossRef 19. BAY 11-7082 Grainger DC, Aiba H, Hurd D, Browning DF, Busby SJ: Transcription factor distribution in Escherichia coli : studies with FNR protein. Nucleic Acids Res 2007,35(1):269–278.PubMedCrossRef 20. Eiglmeier K, Honore N, Iuchi S, Lin EC, Cole ST: Molecular genetic analysis of FNR-dependent promoters. Mol Microbiol 1989,3(7):869–878.PubMedCrossRef 21. He B, Shiau A, Choi KY, Zalkin H, Smith JM: Genes of the Escherichia coli pur regulon are negatively controlled by a repressor-operator interaction. J Bacteriol 1990,172(8):4555–4562.PubMed 22. Spiro S, Guest JR: FNR and its role in oxygen-regulated gene expression in Escherichia coli .

Similar results were obtained with WT MEFs infected with B melit

Similar results were obtained with WT MEFs infected with B. melitensis-mCherry (Figure 5B). However, in this case, we observed a significant decrease (p < 0.01) in the number of bacteria per infected cell but only at 24 h p.i. Next, we examined the impact of a pre-treatment with 3MA on Brucella replication in host cells using the gentamicin survival assay. Our results show that a pre-incubation of WT MEFs with 3MA does not impair the replication of both B. abortus and B. melitensis (Figure 6 A-B). Figure

5 Impact of 3MA on the infection of WT MEFs with B. abortus -mCherry (A) or with B. melitensis- mCherry (B). The number of bacteria per infected cell was measured on at least 57 infected cells coming from two independent experiments. Values represent means ± SEM. Statistical significance #S3I-201 randurls[1|1|,|CHEM1|]# was calculated using the Mann–Whitney Rank Sum Test. # and ## indicate a significant difference with p <0.05 and p <0.01, respectively. NS stands for non significant difference. Figure 6 Impact of 3MA on the infection of WT MEFs with B. abortus S2308 (A) or with B. melitensis 16M (B). Results represent log CFUs (means ± SD) measured at various times postinfection in at least three independent experiments made in triplicates. Discussion KPT-8602 After internalisation, B. abortus is found inside individual vacuoles that interact transiently with endosomes and perhaps lysosomes [6]. Then, Brucella evades the endocytic pathway and reaches its replicative niche,

an check ER-derived compartment, by a still unknown mechanism.

It is also unclear whether Brucella transits through the autophagic pathway before its replication. Based on the appearance of B. abortus in multilamellar structures looking like autophagosomes and on the decrease of its replication rate after autophagy inhibition with 3MA, Pizarro-Cerda et al. [11] proposed that this bacterium passed through the autophagy pathway before reaching its niche of replication [13]. In agreement with this assumption, Guo et al. (2012) noticed that inoculation of macrophages with B. melitensis stimulated autophagy and that a pre-treatment with 3MA reduced its growth rate [22]. In contrast, using macrophages derived from KO mice or HeLa cells incubated in the presence of siRNA targeting the autophagic machinery, Starr et al. [12] showed that B. abortus does not use the conventional macroautophagic pathway either for its intracellular trafficking between the endocytic compartments and the ER derived-vesicles or for its replication [12]. In our study, we sought to compare the fate of B. abortus and B. melitensis in Atg5-deficient MEFs, i.e. in cells that are unable to set up the conventional pathway of macroautophagy even under starvation conditions. Our results show that both Brucella strains are able to invade and replicate in Atg5−/− MEFs, indicating that Atg5 is dispensable for the intracellular survival and replication not only of B. abortus but also of B. melitensis.

Acknowledgments We would like to thank David L Hawksworth for en

Acknowledgments We would like to thank David L. Hawksworth for enabling and continuously supporting this special issue. We are very grateful to all the participating colleagues of our conference and to all the authors and reviewers for their valuable contributions

to this special issue. Finally, we want to acknowledge the German Federal Ministry for the Environment, Nature Conservation and Nuclear Safety (BMU) for funding the conference on “Forest Biodiversity in a Changing Climate: Understanding Conservation Strategies and Policies” see more and the research project “Selonsertib molecular weight Forests and Climate Change” (FKZ 3508 83 0600) through the German Federal Agency for Nature Conservation (BfN). References Berkes F (2007) Understanding uncertainty and reducing vulnerability: lessons from resilience thinking. Nat Hazards 41(2):283–295. doi:10.​1007/​s11069-006-9036-7 CrossRef Buse J, Griebeler

EM, Niehuis M (2013) Rising temperatures explain past immigration of the thermophilic oak-inhabiting beetle Coraebus florentinus (Coleoptera: Buprestidae) in south-west Germany. Biodivers Conserv 22. doi:10.​1007/​s10531-012-0395-y Caparros A, Jacquemont F (2003) Conflicts between biodiversity and carbon sequestration programs: economic and legal implications. Ecol Econ 46(1):143–157. doi:10.​1016/​S0921-8009(03)00138-1 Smad inhibitor CrossRef Chrysopolitou V, Apostolakis A, Avtzis D, Avtzis N, Diamandis S, Kemitzoglou D, Papadimos D, Perlerou C, Tsiaoussi V, Dafis S (2013) Studies on forest health and vegetation changes in Greece under

the effects of climate changes. Biodivers Conserv 22. doi:10.​1007/​s10531-013-0451-2 Easterling DR, Meehl GA, Parmesan C, Changnon SA, Karl TR, Mearns LO (2000) Climate extremes: observations, modeling, and impacts. Science 289(5487):2068–2074. doi:10.​1126/​science.​289.​5487.​2068 PubMedCrossRef Entenmann SK, Schmitt CB (2013) Actors’ PIK-5 perceptions of forest biodiversity values and policy issues related to REDD+ implementation in Peru. Biodivers Conserv 22. doi:10.​1007/​s10531-013-0477-5 Flannigan MD, Krawchuk MA, de Groot WJ, Wotton BM, Gowman LM (2009) Implications of changing climate for global wildland fire. Int J Wildland Fire 18(5):483–507. doi:10.​1071/​Wf08187 CrossRef Freudenberger L, Hobson P, Schluck M, Kreft S, Vohland K, Sommer H, Reichle S, Nowicki C, Barthlott′ W, Ibisch PL (2013) Nature conservation: priority-setting needs a global change. Biodivers Conserv 22. doi:10.​1007/​s10531-012-0428-6 Hampe A, Petit RJ (2005) Conserving biodiversity under climate change: the rear edge matters. Ecol Lett 8(5):461–467. doi:10.​1111/​j.​1461-0248.​2005.​00739.​x PubMedCrossRef Hannah L, Midgley G, Andelman S, Araujo M, Hughes G, Martinez-Meyer E, Pearson R, Williams P (2007) Protected area needs in a changing climate. Front Ecol Environ 5(3):131–138. doi:10.