These points are taken up in various ways by the papers in this special issue. The papers are organized into three clusters. The first four articles focus on the history and evolution of sustainability science and take stock of current challenges to strengthening the science–policy–Selleck C646 society link; the next two articles consider scientific and institutional barriers to the transdisciplinary approach and means to overcome them; the special issue concludes with two articles that focus on the future. The first of these is an overview article that presents quality criteria for developing visions and visioning

in sustainability research and proposes two integrative research project frameworks drawn from complexity theory that illustrate the Fer-1 PKC412 order use of the criteria. The second explores the value of building social–ecological resilience through a case study on applying sustainability science to strengthening social–ecological resilience in recovery efforts in NE Japan. Kajikawa, Tacoa and Yamaguchi revisit the academic landscape of sustainability science that Kajikawa and other colleagues created in 2007

using an analysis of the citation network to provide evidence of the intellectual evolution of sustainability science (see Kajikawa et al. 2007) In the paper for this special issue, the scholars present the results of their research using citation and text (bibliometric) analysis of published articles and applying this to their methodology to develop a profile of sustainability issues addressed by the science. Their results indicate that separated disciplinary-bound research clusters identified in the earlier study are becoming integrated into those studying coupled systems. An encouraging Pyruvate dehydrogenase sign emerging from the analysis is evidence of an increase in recent years (from 2007 to 2009) of attention to socio-ecological systems and a concomitant interest in the social and political/policy components of the issues studied. Moreover, they find that the science is bridging gaps that are left in traditional scientific

research, especially with respect to gaps between social, ecological and economic systems, between diverse disciplines, and between the current state and a sustainable future. This increase suggests that sustainability science, as reflected in the literature, is becoming more concerned with the science–policy–society link that is crucial to moving societies forward on the path to sustainable development. In his critical examination of five transdisciplinary projects in practice, Polk examines why in some cases knowledge co-generated through transdisciplinary approaches does not necessarily result in the ability to influence change in a sustainable direction. This, he finds, is often due to a lack of sufficient attention paid to delivery mechanisms for sustainability research results.

3 10.4 – 9.1 9.1   2.3 2.4   212107_s_at DEAH www.selleckchem.com/products/bay80-6946.html (Asp-Glu-Ala-His) box polypeptide 9 DHX9 – - – - – - – - – 212917_x_at RecQ protein-like (DNA helicase Q1-like) RECQL 10.6 10.7 – 9.5 9.6   2.2 2.3 – 212918_at RecQ protein-like (DNA helicase Q1-like) RECQL – - – - – - – - – 213520_at RecQ protein-like

4 RECQL4 – - – - – - – - – 213647_at DNA2 DNA replication helicase 2-like (yeast) DNA2L 8.6 8.7 8.7 10.2 10.2 10.2 -3.0 -2.8 -2.8 213878_at similar to CG10721-PA LOC642732 – - – - – - – - – 221686_s_at RecQ protein-like 5 RECQL5 – - – - – - – - – Three different methods for data normalization using ACTB, GAPDH, and Affymetrix U-133A housekeeping Selleck STI571 genes, respectively were utilized. Table 3 Expression of DNA polymerase alpha. Probe set Description Gene symbol PT3 Non-PT3 Fold Differences       ACTB GAPDH U133-A ACTB GAPDH U133-A ACTB GAPDH U133-A 204441_s_at Polymerase (DNA directed), alpha 1 POLA1 – - – - – - – - – 204835_at Polymerase (DNA

directed), alpha 1 POLA1 11.7 11.8 11.8 10.1 10.1 10.1 2.9 3.1 3.1 Three different methods for data normalization using ACTB, GAPDH, and Affymetrix U-133A housekeeping genes, respectively were utilized. Comparison Niclosamide of Normalization Techniques Based on the number of transcripts identified as differentially expressed, the three techniques used to normalize the array data could be ordered by the number of

genes identified as differentially expressed as follows: GAPDH (1869 probe sets) > ACTB (1781 probe sets) > U-133A (1478 probe sets). Although the three array normalization methodologies differed in the number of genes defined as down- or up-regulated in expression in PT3 compared to PT1 and NK cell lines, all identified the same 7 up-regulated genes (PCNA, POLD1, RFC3, RFC4, RFC5, RPA1, and RPA2) except RPA1 in normalization using HG-U133A housekeeping genes (Table1). This finding suggested that these seven genes were clearly differentially over-expressed in PT3 versus PT1 and NK cell lines. Verification of microarray results by real-time quantitative PCR As we did the microarray analysis using a single mRNA isolation/cDNA probe analysis, we needed to verify the transcriptional over-expression of these seven genes by real-time quantitative PCR. To determine the optimum amount of cDNA template in initial experiments, we performed undiluted, 1:10 diluted, and 1:100 diluted cDNA template in find more parallel.

The CCL21 gene was PCR amplified with forward primer 5’-GCG CGG GAT CCC ATG GCT CAG ATG ATG AC-3’ and reverse primer 5’-TCA TGT CGA GCT AGC GGG CTC CAG AZ 628 purchase GCG-3’ using PfuTurbo DNA selleck screening library polymerase (Stratagene, La Jolla, CA). A BamHI site (GGATCC) was inserted into the forward primer to be used for ligation to the expression vector. Amplified CCL21 gene was digested with BamHI and NheI and ligated into the T-REx expression vector digested with

BamHI and XbaI. The integrity of the CCL21 expression plasmid (pcDNA4/TO/CCL21) was confirmed by sequencing. Tumor Cell Lines, Manipulations and Implantation TRAMPC2 cells were established from a prostate tumor from a TRAMP mouse and were kindly provided by Norman Greenberg (Baylor College of Medicine, Houston, TX). To generate stably transfected cell lines, TRAMPC2 cells were transfected with the T-REx repressor (TR) and pcDNA4/TO/CCL21 expression check details vectors (Invitrogen, Carlsbad, CA) using Fugene6 (Roche Applied Science, Indianapolis, IN) following the manufacturer’s protocol. Cells were maintained in antibiotic containing media for at least 3 weeks before testing for tetracycline inducible

expression of CCL21 by ELISA. Briefly 1×105 cells from each clone were seeded in 12 well plates containing 1ml of media in duplicate. The following day the media was replaced with fresh media with or without 2mg/ml of tetracycline (Invitrogen, Carlsbad, CA). The assay was performed on the third day based on the manufacturer’s protocol (R and D system, Minneapolis, MN). To establish an orthotopic tumor, mice prostate glands were surgically exposed and injected with 0.05ml of media containing 5×105 tumor cells. Mice were regularly monitored for tumor growth. Mice were treated with 0.02mg/ml of doxycycline (a tetracycline derivative) along with 0.5% sucrose in their drinking water when indicated. All animal protocols were conducted in accordance with National Institute of Health guidelines and were reviewed Orotidine 5′-phosphate decarboxylase and approved by the Institutional Animal Care and Use Committee of Eastern Virginia Medical School. Tumor infiltrating leukocytes (TILs) were isolated from palpable

tumors that were excised, diced and digested enzymatically as previously reported [13]. Cells were then washed to remove enzymes and dead cells were eliminated from the preparation by Ficoll (Isolymph, Gallard-Schlesinger Industries, Carle Place, NY) gradient centrifugation [11]. Single cell suspension of spleens from normal mice and tumor bearing mice were prepared following the procedure for TILs and used as control. To detect metastatic disease in mice with TRAMP tumors, different tissues (lymph nodes, lungs, pancreas and bone marrow) were harvested aseptically and cultured as described previously [14]. In some cases prostate tumors were cultured using the same technique and cells from explanted outgrowths were expanded for re-injection into the prostate gland.

g. at the start

and during (final) sprints. In these occasions, i.e. when exercising above CP, W’ will be reduced. Consequently, a higher W’ can increase performance during tests of longer duration, especially if pacing strategies are implemented. We also found that five bolus intakes on five consecutive days did not result in an increase of T lim beyond the value observed after the first intake. Thus, multiday administration of NaHCO3 did not lead to a cumulative effect on endurance capacity. Accordingly, [HCO3 -], blood pH, and ABE after multiday NaHCO3 administration also remained unchanged relative to the initial rise after the first bolus. The most obvious explanation would be that during each CP-trial Nocodazole concentration a certain amount of NaHCO3 was used, leading to lower values for [HCO3 -], pH and ABE post vs. pre test. During the following 24 h of check details recovery, the body would then be expected to re-establish the resting values.

On the following day, the participants then would start the CP trial at similar (complete recovery) or lower [HCO3 -], blood pH, and ABE (incomplete recovery) relative to the first day, whereby an additional increase in performance would not be expected. Although we did not measure [HCO3 -], pH and ABE before supplementation on the following days, these two described cases can be most likely excluded. The reason for this is that [Na+] also did not increase during the consecutive 5 days MI-503 of NaHCO3 supplementation despite the fact that Na+, unlike HCO3 -, was not used as a buffer during the CP trials, and that the high amount of ingested Na+ could not be used completely through sweating. HAS1 The predicted sweating rate during exercise of 1 dm3∙ h-1 water, with a sweat [Na+] of 50 mEq∙ dm3[34] would have led to a Na+ loss of ~0.36 g. This calculated sweat-induced loss of Na+ corresponds to ~20% of the daily Na+ intake during the placebo intervention. Regarding the substantially higher Na+ intake during the NaHCO3 intervention, the sweat-induced loss of Na+ was negligible during

this intervention. As shown in this study, the NaHCO3 intervention led to an increase in [Na+] and plasma osmolality after the first bolus administration. This increase was counteracted by an expansion in PV. The increase in PV was to such an extent that pre-exercise blood [HCO3 -], pH, and ABE remained constant during the 5 days of testing. This proposed mechanism of PV expansion has already been described by Máttar et al.[35], who showed that plasma [Na+] and plasma osmolality were increased after NaHCO3 injections in acute cardiac resuscitation. Other mechanisms to counteract increases in [Na+] and plasma osmolality comprise a shift of fluid from the intra- to the extramyocellular compartment [36], a stimulation of arginine vasopressin secretion [37], which leads to an intensified water retention from the kidneys [38], and a stimulation of the thirst center whereby more fluid is consumed [37]. In accordance with our results, McNaughton et al.

The usefulness of MLVA was recently demonstrated for the avian major pathogen Chlamydophila psittaci [5]. The objective of the present study was to develop a new typing method based on the detection of VNTRs in the filamentous fungus A. fumigatus, another major avian pathogen. All putative VNTR markers were screened on the whole genome of A. fumigatus

strain Af293. Ten markers were finally selected and used for the typing of a large number of isolates from poultry and their environment in France and China. Methods Strain collection In order to develop a MLVA scheme and choose discriminant VNTR markers, a total number of 57 isolates was selected from our laboratory collection. These isolates were considered as geographically or temporally unrelated. The isolates Fedratinib chemical structure were collected from tissues or from pharyngeal swabs: (i) 49 isolates from different animal species with lesions of aspergillosis in different places in France (n = 48) or Morocco (n = 1); (ii) 3 isolates collected from human cases of aspergillosis at one hospital in Ile-de-France region,

France; (iii) check details 2 isolates collected from healthy birds in 2 poultry farms in France; (iv) 2 isolates from healthy birds in chicken and duck farms in Guangxi province, China; (v) the reference strain CBS 144.89 (Table 1). Table 1 Origin and period of collection for 57 unrelated isolates of Aspergillus fumigatus find more examined in the present study Isolates no Hosts Period of collection Geographic origin S1-S15, S24, S25, S28-S35, S38, S46, S48, S49, S50, S51, S54, S55 Ducks (Anas platyrhynchos), pulmonary aspergillosis 10/2007-04/2008 Poitou-Charentes, France S17, S18 Pigeons (Columba livia), pulmonary aspergillosis 11/2007 Ile de France, France S19, S20, S22, S23, S26, S36, S42, S44, S52, S53, S56 Turkey

(Meleagris gallopavo), pulmonary aspergillosis else 11/2007-04/2008 Poitou-Charentes, France S40, S41 Pheasant (Phasianus colchicus), pulmonary aspergillosis 01/2008 Poitou-Charentes, France E19, E20 Ducks (Anas platyrhynchos), asymptomatic carriage in pharynx 01/2008-04/2008 Sarthe, France D3 Chicken (Gallus gallus), asymptomatic carriage in pharynx 02/2008-03/2008 Guangxi province, China D42 Duck (Anas platyrhynchos), pulmonary aspergillosis 02/2008-03/2008 Guangxi province, China V04M02253 Bustard (Chlamydotis undulata), asymptomatic carriage in trachea 01/2008-04/2008 Morocco H50, H71, H100 Patients, pulmonary aspergillosis 12/2005-04/2008 Ile de France, France CBS 144.

Next, double-distilled water was added and the cells were incubated for 4 h at 25°C to obtain total lysis. The lysates were centrifuged

at 1,400 × g for 5 min, and the supernatant underwent electrophoresis by SDS-PAGE. Proteins in the gel were blotted onto a nylon membrane; membrane strips were incubated with blocking buffer for 4 h at 25°C. Incubation for 1 h with streptavidin-HRP followed. A control containing PbMLSr was revealed with the Catalyzed Signal Amplification (CSA) System kit (DAKO). The negative control was developed with the supernatant of A549 cells after lyses (without incubation with the biotinylated protein). Confocal analysis The cellular localization of the PbMLS was performed as described by Batista et al. [55] and Lenzi et al. [56] for confocal laser scanning microscopy (CLSM). Briefly, the cells growing in different sources of carbon were fixed in 4% paraformaldehyde for 1 h, washed and centrifuged. After permeabilization with Triton find more X-100, the cells were washed in PBS and incubated in blocking solution (2.5% BSA, 1% skim milk, 8% fetal calf serum) for 20 min (Fernandes da Silva, 1988). The diluted (1:100) primary antibody anti-PbMLSr was added overnight at 4°C. After washing three times with PBS, the cells were incubated

with secondary antibody (Alexa Fluor 488 anti-rabbit Molecular Probes 1:700) for 1 hour. Before mounting in 90% glycerol in PBS, adjusted to pH 8.5, containing antifading agent (p-phenylenediamine 1 g/L) (Sigma-Aldrich), the cells were stained with Evans blue (1/10000 in 0.01 M PBS). The specimens were analyzed by laser confocal microscopy (LSM 510-META, Zeiss). Flow cytometry BV-6 in vitro assay analysis All flow cytometry analyses were performed on a BD FACSCanto (BD Biosciences) using an air-cooled argon-ion laser tuned to 488 nm and 115 mW. The flow rate was

kept at approximately 10,000 events (cells), and green fluorescence was amplified logarithmically. Ten thousand events were collected as monoparametric histograms of log fluorescence, as well as list mode data files. The data were analyzed by FACSDiva Software (BD Biosciences) and Origin Software [54]. Enzymatic activity MLS activity was determined as described by Zambuzzi-Carvalho (2009) [30]. Briefly, the enzymatic assay was carried out at room temperature. 25 mg samples were added to 500 ml assay buffer Celecoxib containing 5 mM acetyl-CoA (20 ml) and water to a volume of 980 ml. The reaction had the optical densities read at 232 nm until stabilization. The enzymatic reaction was started by the addition of 100 mM glyoxylate (20 ml). The method is based on the consumption of acetyl-CoA at 232 nm. The activity was calculated considering that one unit at 232 nm is defined as 222 nmols/mg of acetyl-CoA. The specific activities were given in U/mg protein, with U being equal at nmol/min. Statistical analysis Results are expressed as the mean ± SD of the mean of three SGC-CBP30 nmr independent experiments.

). After four MG-132 days the phages 1 × 106 and bacteria (5 × 106) were administered. The mice were bled for the measurement of antibody titer 21 days later. The number of mice was 10 per group. Statistics: CP-P-B- vs CP-P-B+ P = 0.0495; CP-P-B+ vs CP+P-B+ P = 0.0369; CP+P-B+ vs CP+P+B+ P = 0.0001 (ANOVA of Kruskal-Wallis; P = 0.0000). Discussion The results presented in this report demonstrated

not only efficient removal of the bacterial load in infected mice virtually devoid of major functional phagocytes, by prophylactic administration of specific phages, but also revealed accompanying, beneficial effects on the immune system, mediated by S. aureus phage preparation in the described model. It appeared that application of phages in infected mice may accelerate renewal of cells depleted by CP treatment, both of the myelocytic and lymphocytic lineages. The first type of cells has significance in the first-line defense against bacteria as phagocytes and the latter differentiate to mature, immunocompetent cells, giving rise to adaptive, antigen-specific immune response. It is conceivable that the stimulation of hematopoiesis by phages is Selleckchem Elafibranor initiated by destruction of bacterial walls and release of bacterial antigens acting as selleck kinase inhibitor adjuvants for the immune system.

The stimulatory effects of different bacterial antigens on hematopoiesis in CP-immunosuppressed mice were reported by others [33, 34]. The increased stimulation of hematopoiesis in infected, phage-treated (CP+P+B+) mice versus infected (CP+P-B+) mice or mice treated only with phages (CP+P+B-), found in this

study, supports such a notion. In infected mice not treated with CP, the phages elevated the percentage of mature neutrophils (segments) (CP-P+B+ versus CP-P-B+ mice), although not significantly. That phenomenon could represent an additional output of mature neutrophils from the bone marrow reservoir which is particularly large in rodents [35]. The infection of Phosphoglycerate kinase CP-treated mice (CP+P-B+ group) resulted in a characteristic change in the blood picture with appearance of immature and mature neutrophils as well as more immature cells from myelo- and lymphocytic lineages. The proportion of these cell types, including a small contribution of eosinophils and monocytes, significantly increased in mice treated additionally with phages (from 40.4 to 70.2%) (Figure 3). The effective killing of bacteria in the investigated organs, particularly in the liver, where most of the killing takes place [36], probably resulted from the increased number of phagocytes, as shown in this work, although we assume that a contribution of phages in that process is the major one. The bone marrow picture in normal mice showed a significant increase of the mature neutrophils content after infection (CP-P-B+ group), with a reduction in these cells upon phage application that suggests an accelerated export of neutrophils into periphery.

Cell viability and growth were monitored continuously after applying increasing concentrations of the Ltc 1 peptide (0 (cyan), 12.5 (purple), 25 (dark green), 50 (magenta), 100 (orange), 150 (blue), 200 (green), and 250 μM (red)). (C) The effect of the Ltc 1 peptide on buy Ruxolitinib virus replication in infected

cells. Viral particles were labelled with FITC fluorescence dye using indirect immunostaining, and the cell nuclei were stained with Hoechst. The figure shows a significant reduction of viral particles after peptide treatment. (D) Western blot analysis of the DENV2 NS1 protein expression level normalised to beta-actin as a reference cell protein (L1, untreated control; L2, DENV2-infected cells treated with Ltc 1 peptide). Determination of antiviral inhibitory dose Quantitative real-time PCR was used to determine the viral copy numbers in the infected cells after treatment with the Ltc 1 peptide. The infected cells were treated with increasing concentrations of the Ltc 1 peptide

for 24, 48 and 72 h. The Ltc 1 peptide showed dose-dependent inhibition of DENV2 replication in HepG2 cells. However, the results showed insignificant effects for the time points on peptide SAHA HDAC activity (Figure  4). The inhibitory effects of the Ltc 1 peptide were dependent on increasing concentrations of the peptide at the three time points. The Ltc 1 peptide inhibited DENV2 replication at EC50 values of 8.3 ± 1.2 μM for 24 h, 7.6 ± 2.7 μM for 48 h and 6.8 ± 2.5 μM for 72 h (Figure  4). The mode of inhibition The antiviral activity of the Ltc 1 peptide

was selleck kinase inhibitor further verified by plaque formation assay that showed different inhibitory effects of the peptide against virus entry and replication in infected cells. The Ltc 1 peptide showed significant inhibitory effects at a pre-treatment, simultaneous and post-treatment compared to the untreated cells. However, the antiviral activity for the simultaneous and post-treatment was significantly higher than the pre-treatment (Figure  4A). The viral load (pfu/ml) was significantly (p < 0.001) reduced at pre-treatment (4.5 ± 0.6) compared to the untreated cells (6.9 ± 0.5). In addition, a significant decrease (p < 0.0001) in viral load was observed for the simultaneous treatment (0.7 ± 0.3 Gefitinib order vs. 7.2 ± 0.5 control) and post-treatment (1.8 ± 0.7 vs. 6.8 ± 0.6 control) as shown in Figure  5A and 5B. Figure 4 Determination of viral inhibitory dose of the Ltc 1 peptide by RT-qPCR. Serial concentrations of the Ltc 1 peptide (0, 2.5, 5, 10, 20, 40, and 80 μM) were incubated with HepG2 cells infected with DENV for 72 h. The viral RNA was quantified by one-step qRT-PCR. The results showed a dose-dependent reduction in viral copy number after treatment with the Ltc 1 peptide for 24, 48 and 72 h. Figure 5 Mode of action of the Ltc 1 peptide against DENV2 infection.

Nevertheless, it is feasible and has the potential to be developed to a nationwide database. Analysis of a trauma registry as early as six months can lead to useful information which has the potential for long term effects on the progress of trauma research and prevention. Acknowledgements We thank Trauma Services at Royal Perth Hospital,

Perth, WA, Australia for permission to modify and use their data collection forms. This study has been supported by grants from the United Arab Emirates University (Project # 01-07-8-11/03) and the Faculty of Medicine and Health Sciences, UAE University (NP/03/011). References 1. Krug EG, Sharma GK, Lozano R: The global burden of injuries. Am J Public Health 2000, 90:523–526.CrossRefPubMed 2. World Health Organization: [http://​www.​who.​int/​violence_​injury_​prevention/​publications/​road_​traffic/​world_​report/​en/​] 2004 World report on road AMN-107 traffic injury prevention 3. Fikri M, Noor AM, Shaheen H: Preventive Apoptosis inhibitor Medicine Department 1998 Annual Report. Ministry of Health, Abu-Dhabi, UAE 4. Schultz CR, Ford HR, Cassidy LD, et al.: Development of a hospital-based trauma registry in Haiti: an approach for improving injury surveillance in LY3023414 developing and resource-poor settings. J Trauma 2007, 63:1143–1154.CrossRefPubMed 5. Probst C, Paffrath T, Krettek C, et al.:

Comparative update on documentation of trauma in seven national registries. Eur J Trauma 2006, 32:357–365.CrossRef 6. Moore L, Clark D: The value of trauma registries. Injury 2008, 39:686–695.CrossRefPubMed 7. Nwomeh BC, Lowell W, Kable R, et al.: History and development of trauma registry: Lessons from developed to developing countries. World J Emerg Surg 2006, 1:32.CrossRefPubMed 8. Abu-Zidan FM, Lunsjo K, Shaban S, et al.: Establishment of a trauma registry: Experience

from UAE. ANZ J Surg 2005,75(Suppl):A111-A114. 9. Eid HO, Barss P, Adam SH, et al.: Factors affecting anatomical region of Injury, Severity, and Mortality for Road Trauma in a high-income developing country: lessons for Prevention. Injury 2009, 40:703–707.CrossRefPubMed 10. Eid HO, Lunsjo K, Torab FC, et al.: Pre-incident behavior and mechanism of injury in drivers involved in vehicle collisions in the UAE. ANZ J Surg 2008,78(Suppl 1):A143. Glycogen branching enzyme 11. Hefny A, Eid HO, Abu-Zidan FM: Severe Tire Blast Injuries during Servicing. Injury 2009, 40:484–487.CrossRefPubMed 12. Hefny A, Eid HO, Salim K, Abu-Zidan FM: Fatal Tire Blast Injury Causing Bowel Evisceration and Forearm Amputation. NZ Med J 2008.,121(1282): 13. Barss P, Addley K, Grivna M, et al.: Occupational injury in the United Arab Emirates: epidemiology and prevention. Occup Med (Lond) 2009, in press. 14. Cameron PA, Gabbe BJ, McNeil JJ, et al.: The trauma registry as a statewide quality improvement tool. J Trauma 2005, 59:1469–1476.CrossRefPubMed 15. Sanidas EE, Valassiadou KE, Kafetzakis AG, et al.

F. alocis thus seems to be a powerful diagnostic marker organism for periodontal disease. FISH revealed the involvement of F. alocis in numerous structural arrangements that point to its potential role as one

of the architects of structural organisation within periodontal biofilms. Filifactor alocis should be considered an important periodontal pathogen and warrants further research. Acknowledgements We thank Eva Kulik, University of Basel, and Eivind Strøm, University of Oslo, for providing clinical samples, Cindy Hefenbrock and Marie Knüver for excellent technical assistance, Derek Ramsey for proof reading, and Dr. Wolf-Ulrich Klotz for his support. This work was supported by the Sonnenfeld-Stiftung, Berlin, Germany, and by a Rahel-Hirsch selleck compound learn more grant from Charité – Universitätsmedizin to AM. Electronic supplementary material Additional file 1: Optimization of probe FIAL for FISH using the program daime. FISH was performed incubating fixed cells of F. alocis and F. villosus with different hybridization mixes. Signal intensities (Relative fluorescent Units, RU) emitted by F. alocis and F. villosus at different formamide concentrations were calculated from images taken with a fixed exposure time. Due to unspecific binding of FIAL, the light emission of F. villosus cells

remained below 50 RU at every level of formamide. The signal emitted by F. alocis cells was considered sufficient using formamide concentrations of up to 20% (v/v). (PPT 53 KB) References 1. Haffajee AD, Socransky SS: Microbial etiological agents of destructive periodontal diseases. Periodontol 2000 1994, 5:78–111.PubMedCrossRef 2. Kolenbrander

PE, London J: Adhere today, here tomorrow: oral bacterial adherence. J Bacteriol 1993, 175:3247–3252.PubMed 3. Dahlen GG: Black-pigmented gram-negative anaerobes in periodontitis. FEMS Immunol Med Microbiol 1993, 6:181–192.PubMedCrossRef Teicoplanin 4. Fives-Taylor PM, Meyer DH, Mintz KP, Brissette C: Virulence factors of www.selleckchem.com/products/q-vd-oph.html Actinobacillus actinomycetemcomitans. Periodontol 2000 1999, 20:136–167.PubMedCrossRef 5. Cutler CW, Kalmar JR, Genco CA: Pathogenic strategies of the oral anaerobe, Porphyromonas gingivalis. Trends Microbiol 1995, 3:45–51.PubMedCrossRef 6. Sela MN: Role of Treponema denticola in periodontal diseases. Crit Rev Oral Biol Med 2001, 12:399–413.PubMedCrossRef 7. Slots J, Listgarten MA: Bacteroides gingivalis, Bacteroides intermedius and Actinobacillus actinomycetemcomitans in human periodontal diseases. J Clin Periodontol 1988, 15:85–93.PubMedCrossRef 8. Murray PA, French CK: DNA probe detection of periodontal pathogens. In New biotechnology in oral research. Edited by: WM M. Basel: Karger; 1989:33–53. 9. Chuba PJ, Pelz K, Krekeler G, de Isele TS, Gobel U: Synthetic oligodeoxynucleotide probes for the rapid detection of bacteria associated with human periodontitis. J Gen Microbiol 1988, 134:1931–1938.PubMed 10.