catarrhalis. The overall presence of lactoferrin receptors in M. catarrhalis isolates suggests its important role in colonization or infection [32]. In our previous study we demonstrated that exposure of M. catarrhalis to 26°C increases the release of proinflammatory cytokine IL-8 in pharyngeal epithelial cells likely leading to the increased inflammation [10].

Thus, greater local concentrations of IL-8 would promote enhanced recruitment and influx of neutrophils that release lactoferrin from their secondary granules, which contribute to lactoferrin levels both locally and in the circulation [33, 34]. On the other hand, increased expression of M. catarrhalis lactoferrin binding proteins following cold shock would facilitate the binding

and acquisition of iron from lactoferrin to support growth of bacteria in the mucosal environment. NVP-LDE225 It has been shown that supplemental lactoferrin can enhance the virulence of meningococcal infection in mice [35]. In addition to iron acquisition, lactoferrin receptors may provide protection against anti-bacterial cationic peptides (eg, lactoferricin) and reduce complement-mediated killing. The pneumococcal surface protein PspA binds lactoferrin and protects Streptococcus pneumoniae against the antibacterial effect of lactoferricin [26]. The release of LbpB from the cell surface by a NalP protease protects Neisseria meningitidis against bactericidal antibodies [36]. Therefore, increased expression

of lactoferrin receptors and enhanced binding of 26s Proteasome structure lactoferrin on the surface of bacteria following cold shock might be associated with enhanced protection of M. catarrhalis against anti-bacterial cationic peptides and bactericidal antibodies. The level of UspA2 protein that afforded serum resistance in the bactericidal activity assay has been shown to Non-specific serine/threonine protein kinase correlate with increased binding of vitronectin [37]. Our results indicate that cold shock upregulates the UspA2 protein expression and promotes M. catarrhalis binding to vitronectin. Increased UspA2 protein expression at 26°C was not the result of higher copy number of uspA2 mRNA, indicating that post-transciptional mechanisms are involved in upregulation of this protein after cold shock [38]. Cold shock did not influence the serum resistance of O35E strain indicating that M. catarrhalis strains may need to maintain a certain threshold level of UspA2 protein necessary to evade host defenses. Most seroresistant M. catarrhalis strains express at 37°C sufficient levels of UspA2 to mediate serum resistance [37]. It is conceivable that cold shock would increase UspA2 expression and vitronectin binding in M. catarrhalis strains constitutively expressing low levels of UspA2, leading to the enhanced serum resistance. The infant population during the first year of life possesses a substantial proportion of IgD in saliva [39].

Specificity and limit of detection of the fiber-optic sensor The specificity and limit of detection (LOD) of the fiber optic sensor were analyzed

by using MAb-2D12 as capture antibody and Cy5-labeled MAb-2D12 as a reporter. The sensor generated strong signals against L. monocytogenes and L. ivanovii, with a maximum signal of 22,560 pA. In contrast, non-pathogenic Listeria produced see more a maximum signal of 3,000–4,200 pA (Figure  7a), and non-Listeria bacteria, including Salmonella Typhimurium; E. coli O157:H7; and background food contaminant isolates, Staphylococcus aureus, S. epidermidis, Enterobacter cloacae, and Lactococcus lactis[50], produced signals of ~2,500 pA (Figure  7b). Similar results were obtained when MAb-3F8 was used as the capture and MAb-2D12 as the reporter molecule (Figure  7a,b). In the mixed cultures containing L. monocytogenes, L. innocua, and E. coli O157:H7 (~106 CFU/mL of each), the signals for MAb-2D12 and MAb-3F8 were 15,440 ± 1,764 pA and 8,440 ± 569 pA, respectively, which were significantly (P < 0.05) higher than the values obtained for L. innocua (2,725 ± 2,227 pA) or E. coli (1,589 ± 662 pA) alone (Figure  7b). The background control (PBS only) values ranged from 504– 650 pA. Therefore, both fiber-optic sensor configurations, 2D12–2D12 and 3F8–2D12, are highly specific for pathogenic Listeria, and specificity was contributed primarily by anti-InlA MAb-2D12. Other combinations did not produce satisfactory

learn more results (data not shown). Figure 7 Determination of specificity (a, b) and detection limit (c, d) of the fiber-optic sensor using MAb-2D12 (InlA) or MAb-3F8 (p30) as capture antibody and Cy5-conjugated anti-InlA MAb-2D12 as a reporter against (a) Listeria spp. and (b) other bacteria. Culture

concentrations selleck inhibitor were 108 CFU/mL (or ~106 CFU/mL for mixed-culture experiments). Detection limit of the fiber-optic sensor using (c) MAb-2D12 and (d) MAb-3F8 as capture and MAb-2D12 as a reporter against different concentrations of L. monocytogenes or L. ivanovii. Signals (pA) are the mean of three fibers at 30 s. The LOD was also evaluated by using pure cultures of L. monocytogenes and L. ivanovii serially diluted in PBS (Figure  7c and 7d). Using MAb-2D12 as the capture molecule, the signals increased proportionately as the bacterial concentration increased until a cell concentration of 1 × 106 CFU/mL was reached, which gave the maximum signal (22,560 pA), almost reaching the threshold of the Analyte 2000 fluorometer. The lowest cell concentration that was considered positive (within the detection limit) was 3 × 102 CFU/mL for L. monocytogenes (6,252 ± 1,213 pA) and 1 × 103 CFU/mL for L. ivanovii (8,657 ± 4,019 pA). These values were at least 2-fold higher than those produced by the samples with 101 cells or PBS (blank). When MAb-3F8 was used as capture antibody, the LOD for L. monocytogenes (16,156 ± 6,382 pA) and L. ivanovii (13,882 ± 5,250 pA) was ~1 × 105 CFU/mL (Figure  7d).

interrogans Bataviae – - – - Selleckchem Defactinib – + + – + – L. kirschneri Grippotyphosa – - – - – + – - + + The displayed peaks are based on visual comparison of the algorithms analysis results of the software. All strains were screened twice using the QuickClassifier (QC)/Different average and SNN algorithms. The used symbols stand for: no peak found: – peak present: + peak set with high intensity: ++ Table 5 Differentiating peaks based on the statistical analysis of ClinProTools within the species L. borgpetersenii genomospecies peak mass (m/z) representing

the protein size in Dalton 3,759 5,765 5,779 6,388 7,519 7,547 L. borgpetersenii Ballum + – + – + – L. borgpetersenii Javanica + – + – + – L. borgpetersenii Sejroe + – + – + – L. borgpetersenii Saxkoebing – - + + – + L. borgpetersenii Tarassovi + + – + + – The displayed peaks

are based on visual comparison of the algorithm analysis results of the software. All strains were screened twice using the QuickClassifier (QC)/Different average and SNN algorithms. The used symbols stand for: MDV3100 cost no peak found: – peak present: + The additional statistical tool Principal component analysis (PCA) included in ClinProTools was applied to the analyzed datasets to visualize the homogeneity and heterogeneity of the protein spectra. PCA reduces the variables of a complex dataset on the basis of different statistical tests. The reduced datasets, the so-called PCs (principle components) can be displayed in a score plot illustration. Twenty individual protein spectra of the Silibinin L. interrogans strains and the L. kirschneri strain are displayed in three-dimensional PCA in Figure 2. Each dot stands for a displayed protein spectrum. The colors indicate the calculated cluster membership in which each dot represents one measured protein spectrum

profile for each sample. A clear separation of the serovars Pomona and Copenhageni is apparent. Conversely, L. kirschneri serovar Grippotyphosa did not cluster separately in PCA analysis, even if specific peaks could be detected for L. kirschneri in the peak statistics (see Table 4). For the genomospecies L. borgpetersenii the separation of the serovars Saxkoebing, Sejroe and Tarassovi was apparent when PCA was performed (Figure 3). Figure 2 Principle Component Analysis (PCA) of the analyzed strains of the genomospecies. L. interrogans and L. kirschneri using the software tool ClinProTools. Figure 3 Principle Component Analysis (PCA) of the analyzed strains of the genomospecies. L. borgpetersenii using the software tool ClinProTools. Strain confirmation and molecular sequencing Sequence analysis of the 28 leptospiral reference strains was performed on the basis of MLST analysis (Figure 4) and 16S rRNA gene sequencing (Figure 5). Confirmation of the field isolates relied on 16S rRNA gene sequencing. Species identity of all used strains was confirmed. Furthermore, the constructed phylogentic trees (Figures 4 and 5) revealed comparable clustering of the leptospiral strains.

The inhibition

of NF-κB activity by inhibitor of nuclear factor κB α (IκBα) would remarkably decrease the level of YY1, and consequently neither EZH2 nor HDAC1 could be recruited to miR-29 promoter [14]. This study demonstrated that NF-κB might be an upstream regulator of the epigenetic status of miR-29 in skeletal myogenesis. Selleckchem GDC-0068 In addition to these effects in solid tumors, miR-29 deregulation by epigenetic mechanisms can also be found in human hematological cancers. For instance, in acute myeloid leukemia (AML), the transcriptional complex NF-κB/Sp1 can interact with HDAC1 and HDAC3 to form the NF-κB/Sp1/HDAC complex on miR-29b enhancer, which resulted in the silencing of miR-29b. Notably, MYC can directly bind to miR-29b promoter and stimulate the activity of NF-κB/Sp1/HDAC. https://www.selleckchem.com/products/th-302.html Therefore, the down-regulation of miR-29b is MYC-dependent [15]. Interestingly, HDAC inhibition could restore the expression of miR-29b in only one third of chronic lymphocytic leukemia (CLL) samples [16]. For the other two-thirds of CLL cases, the identification of other histone modifications that contribute to epigenetic silencing of miR-29b still needs to

be accomplished. In summary, binding of MYC or NF-κB on the miR-29 promoter seems to be a primary event in miR-29 silencing, and thereby induces the initial step of its chromatin modification. Subsequently, various histone modifying enzymes such as EZH2 and HDACs can be recruited to the miR-29b promoter. These enzymatic effectors might receive signals from their initiator, and then function as an executor of this epigenetic event. Additionally, the transcription factors YY1 and Sp1, which are dispensable in this regulation, might act as bridges that connect the initiator and the executor. Let-7 family Reportedly, the let-7 miRNAs, which target oncogenic Ras and function as tumor suppressors, are

located in fragile genomic regions that are frequently deleted in human cancers [1, 17]. Besides genomic alterations, the let-7 genes could also be regulated by epigenetic mechanisms. MYC induced by H. pylori CagA in gastric cancer cells can suppress the expression of let-7a and let-7c through two epigenetic approaches: (1) MYC stimulates EZH2 expression by reducing its negative regulators, miR-26a and miR-101; (2) MYC interacts with DNMT3B Docetaxel nmr and EZH2 on the let-7 promoter, and consequently the let-7 gene is silenced through both DNA and histone methylation. Accordingly, the Ras pathway is activated to contribute to carcinogenesis [18]. However, in human lung cancers, let-7a-3 was found to be hypomethylated, which is different from its status in normal lung tissues [19], suggesting that differential, and even opposite, epigenetic regulations might take place in the same miRNA according to the cell context. In view of that, exploration into the epigenetic modulation of the let-7 gene family is essential.

The membrane was probed with an anti-SOX9 rabbit antibody (1:2,000 dilution; Millipore) and incubated with goat anti-rabbit immunoglobulin G (1:50,000 dilution; Pierce). Expression of SOX9 was determined with SuperSignal CX-4945 in vivo West Pico Chemiluminescent Substrate (Thermo,

USA) according to the manufacturer’s suggested protocol. The membranes were stripped and reprobed with an anti-actin mouse monoclonal antibody (1:2,000 dilution; Millipore) as a loading control. Immunohistochemistry (IHC) Immunohistochemical analysis was performed to study altered protein expression in 142 human lung cancer tissues. The procedures were carried out in a similar manner to previously described methods [13]. Paraffin-embedded specimens were cut into 4 μm sections and baked Histone Methyltransferase inhibitor at 65°C for 30 minutes. The sections were deparaffinized with xylenes and rehydrated. Sections were submerged into ethylenediaminetetraacetic acid antigenic retrieval buffer and microwaved for antigenic retrieval. The sections were treated with 3% hydrogen peroxide in methanol to quench the endogenous peroxidase activity, followed by incubation in 1% bovine serum albumin to block non-specific binding. Rabbit anti-SOX9 (1:50 dilution; Millipore) was incubated with

the sections at 4°C overnight. Primary antibody was replaced by normal goat serum in the negative controls. After washing, the tissue sections were treated with biotinylated anti-rabbit secondary antibody (Zymed, San Francisco, USA) followed by a further incubation with streptavidin-horseradish

peroxidase complex (Zymed). The tissue sections were immersed in 3-amino-9-ethyl carbazole and counterstained using 10% Mayer’s hematoxylin, dehydrated, and mounted in Crystal Mount (Sigma). The degree of immunostaining of formalin-fixed, paraffin-embedded sections was viewed and scored separately by two independent investigators, who were blinded to the histopathological features and patient details of the samples. Scores were determined by combining the proportion of positively stained tumor cells and the intensity of staining. The scores given by the two independent investigators were averaged for further comparative evaluation of SOX9 expression. The proportion of positively stained tumor cells was staged Dichloromethane dehalogenase as follows: 0 (no positive tumor cells), 1 (<10% positive tumor cells), 2 (10-50% positive tumor cells), and 3 (>50% positive tumor cells). The cells at each intensity of staining were recorded on a scale of 0 (no staining), 1 (weak staining, light yellow), 2 (moderate staining, yellowish brown), and 3 (strong staining, brown). The staining index was calculated as follows: staining index = staining intensity × proportion of positively stained tumor cells. Using this method of assessment, the expression of SOX9 in lung cancers was evaluated using the staining index (scored as 0, 1, 2, 3, 4, 6, or 9).

Furthermore, antibiotic treatment seemed to mask the effects of endosymbiont number on encapsulation response observed in control colonies, where the bacteria favoured the encapsulation response. Positive effects of symbionts on host immune system have been described in the last years. For example, the facultative symbionts of Acyrthosiphon pisum (the pea aphid) confer it resistance to parasitoid attacks [18]. Recently, it has been demonstrated that Wolbachia confer vigorous antiviral protection to Drosophila [19]. The mechanisms by which the resistance is expressed

is still unknown, but in another https://www.selleckchem.com/products/Roscovitine.html example it was showed that symbiotic bacteria could compete directly for space and resources and thus prevent host colonization by pathogens [24, 25]. Encapsulation is the principal physiological response against parasitoids suggesting an important role of the stimulation induced by Blochmannia in the protection against parasites. This strong interaction between symbiotic bacteria and ants may explain the persistence and broad occurrence of symbiotic bacteria in the Camponotus genus. Ants from Camponotus genus are abundant almost learn more everywhere in the world where ants are found, comprising more than 600 described species within an

estimated number greater than 1,000 species [26]. Its large distribution, the diversity of forms and food behaviour and the occurrence on diverse environments make the system Camponotus/Blochmannia an interesting model to study how ecological forces determine symbiont characteristics and how bacteria determine the ant traits. For example, it is interesting to determine how genetic differences found among different species of Blochmannia could be related to host ecological characteristics. The social

habits of the ants make them particularly vulnerable to several parasites and parasitoids. Phoridae flies are frequently found around Camponotus nests and their influence is fundamental in regulating the ant communities [27]. So, it can be expected that Camponotus species more exposed to Phoridae attack should harbour more bacteria. The physiological selleck chemicals mechanism linking bacterial amount and encapsulation response remains unknown. Although the better workers “”quality”" due to extra nutrients furnished by bacteria is the more probable explanation, direct production of biomolecules in stress situation should not be excluded. An efficient immune system is a major trait allowing the existence of social insect colonies with thousand of individuals, genetically related [28], living close together, constantly exposed to parasitic disease risks. Competition in the first stages of colony growth constitutes also a great challenge to reach the reproductive stage.

% WC composite was obtained at 1,350°C for 2 min at 30 MPa. The best combination of mechanical properties was obtained for a 2 mol.% Y2O3-stabilized ZrO2 composite with 20 wt.% WC, obtained by electroconsolidation at 1,350°C, combining a hardness of 16.5 GPa and a fracture toughness of 8.5 MPa m1/2. Acknowledgements We thank the Research Centre of Constructional Ceramics and The Engineering Prototyping (Russia) for research assistance and for providing the ZrO2 nanopowder synthesized from Ukrainian raw materials, using its developed technology. selleck References 1. Basu B, Lee JH, Kim DY: Development

of WC-ZrO 2 nanocomposites by spark plasma sintering. J Am Ceram Soc 2004,87(2):317–319. 10.1111/j.1551-2916.2004.00317.xCrossRef 2. Malek O, Lauwers B, Perez Y, Baets P, Vleugels J: Processing of ultrafine ZrO 2 toughened

WC composites. J Eur Ceram Soc 2009,29(16):3371–3378. 10.1016/j.jeurceramsoc.2009.07.013CrossRef 3. Pedzich Z, Haberko K, Piekarczyk J, Faryna M, Litynska L: Zirconia matrix-tungsten carbide particulate composites manufactured by hot-pressing technique. Mater Lett 1998, 36:70–75. 10.1016/S0167-577X(98)00010-XCrossRef 4. Anstis GR, Chantikul P, Lawn BR, Marshall DB: A critical evaluation of indentation techniques for measuring fracture toughness: I. Direct crack measurements. J Eur Ceram Soc 1981, 64:533. 10.1111/j.1151-2916.1981.tb10320.xCrossRef 5. Lange FF: Transformation-toughened ZrO 2 correlations between grain size control and composition

in the system ZrO 2 -Y 2 O 3 . J Am Ceram Soc 1986,69(3):40–242. 6. Anné G, Put S, Vanmeensel K, Jiang D, Vleugels Selleck GSK2245840 J, Van der Biest O: Hard, tough and strong ZrO 2 -WC composites from nanosized powders. J Eur Ceram Soc 2005,25(1):55–63. 10.1016/j.jeurceramsoc.2004.01.015CrossRef Competing interests The authors declare that they have no (-)-p-Bromotetramisole Oxalate competing interests. Authors’ contributions EG and OM were the principal investigators of this study. EG investigated the mechanical properties. OM investigated the structure and performed full factorial experiment for technology of hot pressing with direct transmission of high amperage current. VC prepared the experiment, carried out the X-ray analysis, and analyzed the results. All authors read and approved the final manuscript.”
“Background Bionic superhydrophobic (self-cleaning) surfaces with micrometer-nanometer-scale binary structure (MNBS) have aroused great interest of science and engineering fields [1–3], which can be attributed to their potential application prospects such as drag reduction on ship hulls [4], anti-biofouling in maritime industry [5], and anti-icing for power transmission [6]. Their superhydrophobicity (a water contact angle (WCA) larger than 150° and a water sliding angle (WSA) less than 10°) strongly depends on MNBS structure [7, 8].

His eldest brother Krishnaji (1922–1997), who was a

professor of Physics at the University of Allahabad, was his mentor and responsible for shaping his life. For a wonderful Tribute to Professor Krishnaji, see Govindjee and Srivastava (2010). Govindjee received his BSc in 1952 in Chemistry, Botany and Zoology, and his MSc in 1954 in Botany (specializing in Plant Physiology), both from the University of Allahabad. He obtained his PhD in Biophysics in 1960 from the University of Illinois at Urbana-Champaign, Selleckchem Autophagy inhibitor where he studied under two pioneers: Robert Emerson (Sep. 1956–Feb. 1959) and Eugene Rabinowitch (March 1959–Sep. 1960). He has held the following positions: 1960–1961: United States Public Health Service Postdoctoral Fellow; 1961–1965: Assistant Professor of Botany; 1965–1969: Associate Professor of Botany and of Biophysics; 1969–1999: Professor of Biophysics and Plant Biology; 1999–present: Professor Emeritus of Biochemistry, Biophysics and Plant Biology. His collaborative spirit, his teaching spirit and other activities are described in Eaton-Rye (2007a). I recommend the readers to see a large collection of photographs of Govindjee’s younger days as well buy OICR-9429 as photographs of most of his PhD students and his postdoctoral associates in Eaton-Rye (2007b). In addition, Eaton-Rye (2007b) has a list of

PhD theses of all his students as well as names of his 303 collaborators. Since then, this number has risen to at least 350 (see Appendix 1). It shows how he is constantly interacting with many around the World. I suspect this keeps him young. Not surprisingly, beyond his ongoing research and educational activities, Govindjee enjoys recognizing students at conferences by giving them book awards (see e.g., Moore et al. 2012), and/or congratulating those

who receive the Govindjee and Rajni Govindjee Awards for Excellence in Biological Sciences (see e.g., Fig. 2 for pictures of the 2013 Awardees). An example of his recognition of his past students is his writing recollections of one of his distinguished students Tom Wydrzynski (see Govindjee 2008). Fig. 2 Photographs of five 2013 Govindjee and Rajni Govindjee Awardees for Excellence in Biological Science. Top Left: Left to Oxymatrine right: Robert Koester, Rebecca Slattery; the plaque for Eugene Rabinowitch & Robert Emerson; and Govindjee. Top Right: Adriana Corrales Osorio showing her Award certificate; Bottom: Left to right: Samantha B. Primer and Robert Van Buren holding the current Z-Scheme of photosynthesis; and Govindjee; in the background is the 1965 Z-Scheme of Govindjee. [We note that the Z-scheme designed by him and Wilbert Veit is being distributed by the two free around the World for education purposes. See Fig. 4 in Orr and Govindjee (2013); also see http://​www.​life.​illinois.​edu/​govindjee/​photoweb/​subjects.​html#Light-reactions. Photographs of the 2012 and earlier Govindjee and Rajni Govindjee Awardees for Excellence in Biological Science are at (or linked from it): http://​www.​life.​illinois.

Findings on the relation between education and perceived risk concluded that women with high school or less education were more likely to be either unaware of their risk or overestimate their risk, whereas women with college education STI571 datasheet were less likely to have an optimistic bias [14]. The role of religion in health care decisions and perceived risk among people at increased genetic risk has not been deeply investigated yet. It exists a

certain kind of religious fatalism (a belief that some issues are beyond human control but just in God’s hands) that may influence the subjects’ conceptions of how disease occurs and of how much they can be at risk for developing a particular disease based on family history [16, 17]. On the basis of this kind of fatalism we may hypothesize that the perception of the risk is lower for subjects with high spirituality, as demonstrate by JM Quillin research [17]. Furthermore many studies focused on the role played by psychological distress levels and by the personal and family history of tumour in filtering, modifying and completing CH5183284 molecular weight relevant information, concerning the objective risk, affecting in this way the risk perception of developing the disease[11, 12, 14, 18]. As regards the relationship between

the psychological distress and the risk perception findings are opposing. In fact several studies revealed a correlation between high distress levels and high risk perception, while few researches showed no correlation between these two variables [14, 18]. As far as the family history of tumour is concerned, women with a personal and a family history of cancer usually perceived their risk of developing the disease as higher than that of other women. Nevertheless, comparing the risk perception with an objective estimation Morin Hydrate of the risk (Claus,

Gail or BRCA-PRO models), the women affected by cancer and with a family history of tumour are more accurate in their risk estimation than women with a family history of tumour but healthy [11, 12]. Women involved in several studies that revealed an overestimation of the risk perception are usually referred by an affected relative or by health care setting, while the studies that found an underestimation of the risk perception involved women referred by the community. The importance of risk perception in affecting the decisional making process of the counselee and the relationship between the risk perception and other psychological variables are key issues in the research on genetic counselling across different countries. Nevertheless, in Italy, the risk perception has been little studied and counselors still miss relevant information like: how the risk perception is spread on Italian population, how the risk is associated to other psycho-social variables and if the risk perception is accurate or not compared to objective methods of risk estimate[19].

g., vimentin) and gain of a fibroblastoid morphology together with an increased invasive potential have been described in oral squamous cell carcinoma cell lines [17, 18]. Furthermore, down-regulation of E-cadherin expression has been recently associated with poor prognosis in oral squamous cell carcinoma patients [19]. Finally, transforming growth factor-β is considered as playing a key role in the epithelial-mesenchymal transition process as well [11–13]. In our previous study using a 4-nitroquinoline 1-oxide-induced rat tongue carcinoma model, we showed that the appearance

of SMF was closely associated with the development of carcinoma but not with pre-malignant lesions [20]. Furthermore, on an ultrastructural level, we showed that the carcinoma cells, but not their normal counterparts, acquired cytoplasmic microfilaments that were consistent with contractile microfilaments both in appearance

and organization [21]. These BMS202 in vitro events reflect the morphological modifications occurring within the malignant cells, approaching smooth muscle differentiation, probably as part of the epithelial-mesenchymal transition process. The purpose of the present study was to examine the changes in the occurrence of SMF in tongue epithelial lesions with malignant potential (hyperplasia and dysplasia) and in squamous cell carcinoma, selleck chemicals and to assess the expression of transforming growth factor-β in cases of carcinoma. In addition, we attempted to identify the presence of carcinoma cells that co-express epithelial

membrane antigen and α-smooth muscle actin as a reflection of the epithelial-mesenchymal transition process using a double immunostaining method, which was not previously reported in studies on oral cancer in this context. Materials and Methods Study Group Study Population Records of 22 cases of squamous cell carcinoma of the tongue and 39 cases of premalignant lesions of the tongue consisting of hyperplasia (N = 16), mild dysplasia (N = 12), and moderate-to-severe dysplasia (N = 11) were retrieved from the files of the Department of Oral Pathology, School of Dental Medicine, Tel-Aviv University and Institute of Pathology, The Chaim Sheba Medical Center, Tel Hashomer. Diagnoses Lck were reevaluated and classified by two oral pathologists (MV and DD) according to the World Health Organization classification of head and neck tumors [22]. This study was approved by the Helsinki committee of the Sheba Medical Center. Immunohistochemical Stains Three µ wide sections had been cut from the 61 blocks containing the biopsy specimens of the study cases. They were mounted on positive-charged microscope slides (OptiplusTM, Biogenex, San Ramon, CA, USA), dewaxed in xylene, dehydrated in ethanol, rinsed in distilled water, placed in 3% H2O2, and rinsed again in distilled water. The slides were placed in citrate buffer solution, pH=6, in a microwave oven at 92°C for 10 min for retrieval of antigens.