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Information about Laelapidae mites infesting small rodents is scarce. In this study, we investigated morphological and molecular methods for identifying Laelaps jettmari mites, as well as carrying out phylogenetic analysis of these mites using nuclear rRNA 28S region (domains 1-3). Mites were first identified morphologically under the microscope. Afterwards, DNA was extracted using 2.5 % NH4OH solution and the rRNA 28S region (domains 1-3) was amplified by conventional PCR using the primers 43F and 929R. Then the PCR products were analysed by horizontal electrophoresis on 1.5% agarose gel and the positive samples were sent for sequencing to the sequencing centre in Macrogen, Amsterdam (Netherlands). After getting the sequence of L. jettmari, it was uploaded to MEGA X software for analysing it using evolutionary principles. The results of the study suggest that there is no difference between L. jettmari sequences collected from various rodent hosts in different geographical locations. The L. jettmari sequence obtained in this study and the sequences of Laelaps agilis collected from GenBank presented the shortest interspecific genetic distances of 28S rRNA region (domains 1-3), showing the slight evolutionary distance between these species. This paper gives novel molecular information for the identification and phylogenetic analysis of L.jettmari mites based on 28S rRNA region.

In the present study results related to the in vivo administration of Natural Deep Eutectic Solvents (NADES)-solubilized berberine are reported for the first time. NADES are mixtures of small natural compounds having a melting point significantly lower than that of any individual component. Such solvents have gained much attention of the scientific community in the green chemistry area, being considered useful alternatives to common organic solvents. NADES can be used also as administration vehicles, and this can be attractive for nutraceutical products when eutectics are formed with food grade ingredients. In this work, different NADES were prepared using mainly food grade constituents and were tested as solvents for the alkaloid berberine. Three selected NADES/berberine solutions and an aqueous suspension were orally administered to mice with in dose of 50 mg/Kg. Blood levels of berberine were measured by a LC-MS/MS method. The pharmacokinetic analysis revealed a 2-20 fold increase in blood concentration of NADES/berberine with significant changes in pharmacokinetic profile. Natural Deep Eutectic Solvents may thus be considered attractive solubilizing agents and may also play a role in the increase of absorption of poorly bioavailable natural products such as berberine.

Patients with mismatch repair (MMR)-deficient colorectal cancer (CRC) have a more favorable prognosis than patients with tumors with intact MMR. In order to obtain further insights on the reasons for this different outcome, we investigated the interplay between MMR genes and TLR4/MyD88 signaling. The cancer genome atlas (TCGA) databases were selected to predict the differential expression of TLR4 in colon cancer and its correlation with MMR genes. Moreover, the expression of MMR genes and TLR4 was evaluated by immunohistochemistry in 113 CRC samples and a cohort of 63 patients was used to assess TLR4 mRNA expression and MLH1 epigenetic silencing status. In vitro, the effect of MLH1 knockdown on TLR4 expression was quantified by Real Time PCR. TLR4 expression resulted dependent on MMR status and directly correlated to MLH1 expression. In vitro, MLH1 silencing decreased TLR4 expression. These observations may reflect the better prognosis and the chemoresistance of patients with CRC and MMR defects.

PURPOSE To compare the flexural properties and the adhesion of Lactobacillus salivarius (LS), Streptococcus mutans (SM), and Candida albicans (CA) on heat-polymerized (CV), CAD-CAM milled (CAD), or 3D-printed (3D) Poly (methylmethacrylate) (PMMA). METHODS Ultimate Flexural Strength (UFS), Flexural Strain (FS) (%) at Flexural Strength, and Flexural Modulus (FM) of specimens (65.0��10.0��3.3 mm) from each PMMA group (n=6) were calculated by using the 3-point bending test. The surface roughness profiles (R) were measured before and after polishing with a contact profilometer. LS, SM, and CA adhesion on PMMA specimens (n=18) (10 mm in diameter, 3 mm in height) was assessed after 90 minutes and 16 hours by using scanning electron microscopy. The Kruskal-Wallis test with post hoc analysis was performed to compare the groups (alpha=0.05). RESULTS Mean UFS values were 80.79��7.64 MPa for CV, 110.23��5.03 MPa for CAD, and 87.34��6.39 MPa for 3D. Mean FS values were 4.37��1.04% for CV, 4.71��0.62% for CAD, and 6.19��0.13 % for 3D. Mean FM values were 2542��301 MPa for CV, 3435��346 MPa for CAD, and 2371��197 MPa for 3D. CAD had the lowest average R value (0.29��0.16 ��m) before polishing, and bacterial adhesion after 90 minutes of incubation. R value and microbial adhesion were not different amongst groups after polishing and 16 hours of incubation, respectively. CONCLUSION The CAD group displayed the best flexural properties, except for FS, the lowest roughness before polishing and bacterial adhesion after 90 minutes of incubation. All tested PMMAs had similar surface roughness after polishing, and microbial adhesion after 16 hours of incubation.

Twelve strains of Leuconostoc carnosum from meat products were investigated in terms of biochemical, physiological, and functional properties. The spectrum of sugars fermented by L. carnosum strains was limited to few mono- and disaccharides, consistently with the natural habitats of the species, including meat and fermented vegetables. The strains were able to grow from 4 to 37°C with an optimum of approximately 32.5°C. The ability to grow at temperatures compatible with refrigeration and in presence of up to 60 g/L NaCl explains the high loads of L. carnosum frequently described in many meat-based products. Six strains produced exopolysaccharides, causing a ropy phenotype of colonies, according to the potential involvement on L. carnosum in the appearance of slime in packed meat products. On the other side, the study provides evidence of a potential protective role of L. carnosum WC0321 and L. carnosum WC0323 against Listeria monocytogenes, consistently with the presence in these strains of the genes encoding leucocin B. Some meat-based products intended to be consumed without cooking may harbor up to 108 CFU/g of L. carnosum; therefore, we investigated the potential impact of this load on health. No strains survived the treatment with simulated gastric juice. Three selected strains were challenged for the capability to colonize a mouse model and their immunomodulatory properties were investigated. The strains did not colonize the intestine of mice during 10 days of daily dietary administration. Intriguingly, despite the loss of viability during the gastrointestinal transit, the strains exhibited different immunomodulatory effect on the maturation of dendritic cells in vivo, the extent of which correlated to the production of exopolysaccharides. The ability to stimulate the mucosal associated immune system in such probiotic-like manner, the general absence of antibiotic resistance genes, and the lack of the biosynthetic pathways for biogenic amines should reassure on the safety of this species, with potential for exploitation of selected starters.

Esta dissertação de mestrado profissional teve como objeto o estudo da interação da Brigada Militar em grupos de segurança formados no aplicativo WhatsAppcom a participação da comunidade civil. Através da teoria da Representação Social de Maria Stella Grossi Porto, buscou-se compreender Segurança Cidadã pelo senso comum presente nas interações.Diante do ineditismo deste campo virtual de pesquisa, e a fim de melhor respaldar os resultados, outros meios de pesquisas foram adotados para a confirmar os pensamentos e opiniões coletadas: como grupos focais e entrevistas.Assim, diante da inexistência de regulamentação e padronização desta forma de interação pela Instituição, os resultados serviram para ofertar ao final, um modelo de instrução normativa e uma sugestão para orientar o pré-estabelecimento dos grupos, a fim de facilitar a comunicação entre polícia e comunidade. Dessa forma, padronizar e contribuir para uma construção e implementação de uma polícia humanizada no âmbito da filosofia do Policiamento Comunitário. Esta tesis de maestría profesional tuvo como objetivo estudiar la interacción de la Brigada Militar en los grupos de seguridad formados en la aplicación WhatsApp con la participación de la comunidad civil. A través de la teoría de la Representación Social de María Stella Grossi Porto, buscamos entender la Seguridad Ciudadana a través del sentido común presente en las interacciones. Ante el carácter inédito de este campo virtual de investigación, y con el fin de sustentar mejor los resultados, se adoptaron otros medios de investigación para confirmar los pensamientos y opiniones recogidos: como grupos focales y entrevistas. Así, ante la inexistencia de regulación y estandarización de esta forma de interacción por parte de la Institución, los resultados sirvieron para ofrecer al final, un modelo de instrucción normativa y una sugerencia para orientar el preestablecimiento de los grupos, con el fin de facilitar la comunicación entre policía y comunidad. De esta manera, estandarizar y contribuir a la construcción e implementación de una policía humanizada en el ámbito de la filosofía de Policía Comunitaria.

Background: The expression and localization of transforming growth factor-β (TGF-β) pathway proteins in different compartments of the lower airways of patients with stable COPD is unclear. We aimed to determine TGF-β pathway protein expression in patients with stable COPD. Methods: The expression and localization of TGF-β pathway components was measured in the bronchial mucosa and peripheral lungs of patients with stable COPD (n = 44), control smokers with normal lung function (n = 24), and control nonsmoking subjects (n = 11) using immunohistochemical analysis. Results: TGF-β1, TGF-β3, and connective tissue growth factor expression were significantly decreased in the bronchiolar epithelium, with TGF-β1 also decreased in alveolar macrophages, in patients with stable COPD compared with control smokers with normal lung function. TGF-β3 expression was increased in the bronchial lamina propria of both control smokers with normal lung function and smokers with mild/moderate stable COPD compared with control nonsmokers and correlated significantly with pack-years of smoking. However, TGF-β3+cells decreased in patients with severe/very severe COPD compared with control smokers. Latent TGF-β binding protein 1 expression was increased in the bronchial lamina propria in subjects with stable COPD of all severities compared with control smokers with normal lung function. Bone morphogenetic protein and activin membrane-bound inhibitor expression (BAMBI) in the bronchial mucosa was significantly increased in patients with stable COPD of all severities compared with control subjects. No other significant differences were observed between groups for all the other molecules studied in the bronchial mucosa and peripheral lung. Conclusions: Expression of TGF-βs and their regulatory proteins is distinct within different lower airway compartments in stable COPD. Selective reduction in TGF-β1 and enhanced BAMBI expression may be associated with the increase in autoimmunity in COPD.

Our aim was to identify new multi-target compounds endowed with both anti-inflammatory and anti-bacterial activities for treatment of human infections. Diflunisal, a nonsteroidal anti-inflammatory agent, has recently been repurposed for its anti-virulence properties against methicillin-resistant Staphylococcus aureus. Effective synthesis of some aza-analogs of the anti-inflammatory drug diflunisal was carried out following the route involving key oxazole intermediates to obtain o- and m-hydroxypyridinecarboxylic acid derivatives. The newly synthesized diflunisal aza-analogs did not exhibit cytotoxic activity up to 80 μM and some of them exhibited anti-inflammatory activities, decreasing the levels of pro-inflammatory cytokines and prostaglandins induced by bacterial lipopolysaccharide in human primary macrophages. Ten of the diflunisal aza-analogs were found to have interesting antibacterial activity, sensitizing S. aureus, Streptococcus pyogenes, Enterococcus faecium, and Pseudomonas aeruginosa to the antibacterial effects of beta-lactam antibiotics and protein synthesis inhibitors.

An in-vitro and ex-vivo study is presented aimed at elucidating the role or Reactive Oxygen and Nitrogen Species (RONS) in promoting a selective killing of cancer cells by cold plasma treatment. The investigation is, in particular, dedicated to the possibility of emphasizing the selectivity towards cancer cells by combining the plasma treatment with the effect of a molecule known to enhance intracellular ROS production. Lung carcinoma cell lines and cultured primary cells isolated from surgical samples of laryngeal and lung cancers as well as healthy tissue counterparts were treated with an indirect plasma source, in which a helium flow mixed with ambient air in the region between two brass grids is ionized by means of a RF voltage[1]. The helium enriched with active chemical species is then sent to the substrate to be treated. As already reported, this kind of treatment induces an increase of endogenous Reactive Oxygen Species (ROS) in eukaryotic human cells [2]. In the present study, induced ROS generation was confirmed, but a markedly higher increase was found in cancer cells than in healthy ones. The same effect was observed for intracellular nitric oxide (NO). Moreover, incubating the cells with antimycin A (AMA), a molecule known to increase ROS production [3], an amplification of the effect, both for ROS and NO, could be obtained,. The selective increase in endogenous RONS was associated to increased expression of hypoxia-inducible factor (HIF)-a, an oxygen-sensitive transcriptional activator, and to a higher apoptosis in cancer cells than of their healthy counterparts. Incubation with AMA emphasizes these effects. Overall, these results confirm the importance of the role played by RONS in plasma-based cancer treatment, and indicate that the combination with chemotherapeutic drugs could represent a useful tool to enhance the selective effect induced by the plasma treatment.

Autophagy is involved in many biological aspects, including cell survival and death, innate and adaptive immunity and cancer. An involvement of autophagy is also reported in some inflammatory diseases such as asthma. In the present study we explored the role of autophagy in vernal keratoconjunctivitis (VKC), a severe inflammatory disease mainly found in children and adolescents. Autophagy and apoptosis markers (LC3A, LC3B, Beclin-1, cathepsin B, BCL-2, BAX, caspase 3) expression in conjunctival biopsies from 9 active VKC patients and 9 healthy age matched normal subjects were analyzed using immunohistochemistry and qPCR techniques. Conjunctival cells cultures were treated with inflammatory stimuli (IL-1b, histamine, IL-4, TNFa) and analysed by western blotting for autophagy markers expression. LC3B, Catepsin D and B and Beclin-1expression strongly increased in the stroma of VKC whereas the epithelium was consistently negative for all of the molecules studied but positive for Beclin-1 in VKC. qPCR analysis demonstrated a similar mRNAs expression in VKC and normal subjects. In “in vitro” experiments autophagy induction revealed that only LC3B expression was changed in conjunctival fibroblasts by inflammatory stimuli. In particular, both LC3BI, the LC3B free form, and LC3BII, the phosphatidyl-ethanolamine-conjugated form, involved in the autophagosome formation, were decreased in fibroblast cultures at 24h after TNFα stimulation. However, since LC3B-II is normally degraded by lysosomes and the total amount of LC3B-II depends on the balance between its formation and degradation, we analyzed the expression of LC3B-II in the presence and absence of chloroquine, an inhibitor of lysosomal degradation. We found a significant increased amount of LC3BII compared to the control, indicating an over-expression of this protein in stimulated fibroblasts that is quickly damped by its degradation. Since one of the key steps in autophagy is the conversion of LC3B from LC3B-I to LC3B-II, our results suggest that autophagy may be involved in the pathogenesis of VKC., Italian Journal of Anatomy and Embryology, Vol. 121, No. 1 (Supplement) 2016

Tumor necrosis factor alpha (TNFα) is one of the main mediators of inflammatory response in many pathological diseases, involved in a widespread biological functions, including autophagy. Previous data obtained in our laboratory demonstrated that TNFα and some autophagy markers (which markers please indicate) are overexpressed in a severe inflammatory disease such as vernal keratoconjunctivitis (VKC). In the present study we explored the role of TNFα in the induction of autophagy in VKC, using an in vitro model. Primary conjunctival cell cultures were treated with TNFa and analysed by qPCR and western blotting for expression of some autophagy and lysosomial markers at 4, 10 and 24 hours after exposure. qPCR results demonstrated that LC3B, Beclin-1, LAMP1 and p62 strongly increased from 4 to 24 hours, whereas the expression of Catepsin D, a protein implicated in lysosomial apototic pathway, was comparable to that of untreated control. Western blotting analysis revealed lipidation of LC3B quantified as an increased LC3BII/LC3BI ratio. Moreover, double immunofluorescence for Cathepsin D and LAMP1 showed that Cathepsin D was localized within the lysosomes at 4, 10, 24 hours after cell exposure to inflammatory stimuli. In conclusion, our data demonstrated that TNFα significantly induce in VKC LC3B lipidation, LC3BII/LC3BI ratio and p62 (qPCR) in the cells exposed to inflammatory stimuli which shows possible activation of autophagy pathway., Italian Journal of Anatomy and Embryology, Vol. 122, No. 1 (Supplement) 2017

Plasma medicine is an emerging application of low-temperature, atmospheric pressure plasmas to the treatment of several medical conditions. These plasmas are usually produced ionizing a noble gas flow with RF or pulsed high voltage. The presence in the flow of a fraction of air, or specific gases, causes the production of reactive chemical species which interact with the substrate to which the flow is applied. Metastable states of the noble gas also contribute to the interaction, through Penning ionization. The low power used in these plasma sources ensures that the flow remains at almost ambient temperature, so that no thermal effects are induced and living tissues can be treated in a harmless way. The most straightforward application is disinfection, thanks to the effect of the reactive species on the pathogens, especially bacteria, which lack the mechanisms of defense against oxidative stress which allow survival of the eukaryotic cells composing the substrate. Further applications rely on the ability of the activation of cellular processes, such as the intracellular generation of Reactive Oxygen Species (ROS), which can lead to therapeutic effects, including accelerated wound healing and cancer cell selective killing. In this contribution we present the advances made by our group in this field, showing experimental results concerning an indirect plasma treatment performed using a plasma source patented by our group. The targeted applications are the treatment of corneal infections (keratitis) and cancer treatment.

The aim of the study was to analyse the role of heat shock proteins (HSPs) in vernal keratoconjunctivitis (VKC), a recurrent allergic ocular inflammatory disease. We evaluated the expression of some HSPs (Hsp10, Hsp27, Hsp40, Hsp60, Hsp70, Hsp90) in the mucosal biopsies of VKC patients by immunohistochemistry, and in conjunctival cells cultures treated with inflammatory stimuli (IL-1β, histamine, IL-4, TNFβ, UVB irradiation) by western blotting. Immunohistochemical analysis revealed that Hsp10, Hsp27, Hsp40, Hsp70 and Hsp90 expression was significantly increased in VKC whereas the Hsp60 level was unaltered. In vitro induction by inflammatory stimuli in Chang epithelial conjunctival cells revealed that Hsp70 protein expression was significantly increased in epithelial cells line after 4-10 h from histamine and IL-4 stimulation. The same molecule was also overexpressed in conjunctival fibroblast cultures after TNFβ treatment. Hsp90 protein level was increased in the same cell cultures by IL-1β at 4-10-24 h. The Hsp40 protein expression was increased both in epithelial and fibroblast cultures induced by all inflammatory stimuli. Moreover, UVB irradiation significantly increased Hsp90 expression in primary fibroblast culture and Hsp27 in conjunctival epithelial cells after 10 hours. These results indicate that HSPs levels increase in VKC. In particular, Hsp40 expression is up-regulated by all the typical inflammatory stimuli involved in VKC pathogenesis. The specific role of each one of these chaperonins to further induce or counteract inflammation need to be further investigated., Italian Journal of Anatomy and Embryology, Vol 119, No 1 (Supplement) 2014

The role of MAPK kinases in inducing the inflammatory response in the airways of chronic obstructive pulmonary diseases (COPD) patients is incompletely studied. Objectives: To investigate the expression of activated MAPK kinases in bronchial biopsies of COPD patients and the MAPK kinase bronchial epithelial response to oxidative and inflammatory stimuli related to COPD. Expression of phospho(p)-p38, p-JNK1 and p-ERK1/2 was measured in the bronchial mucosa using immunohistochemistry in patients with mild/moderate (n=17), severe/very severe (n=16) stable COPD, control smokers (n=16), control non smokers (n=9) and in a group with mild asthma (n=9). 16HBE cells, challenged with oxidative and inflammatory stimuli, were also studied for IL-8 and MAPK kinases mRNA production. P-p38 was the most expressed MAPK kinase in the bronchial mucosa of all subjects. No significant differences were observed for immune-expression of p-p38, p-JNK and p-ERK1/2 between COPD and control subjects. 16HBE cells treated with H2O2, cytomix (TNFα+IL- 1β+IFNγ) and Lipopolysaccharide (LPS) up-regulated IL-8 mRNA production at 1h or 2h after treatments. P38α mRNA was significantly increased after H2O2 and LPS. JNK1 and ERK1 mRNA were not significantly increased after H2O2, cytomix or LPS treatments. Blocking p38α activity IL-8 mRNA production was not changed at 1h, 2h and 4h after H2O2 or LPS challenge. P-p38 immune-positivity is prevalent in the bronchial mucosa of COPD and asthmatic patients and p38 mRNA is increased after bronchial epithelial challenges suggesting a relevant role for this MAPK kinase in the induction of bronchial inflammation in COPD and asthma., Italian Journal of Anatomy and Embryology, Vol 119, No 1 (Supplement) 2014

Cold atmospheric pressure plasma has been proposed for sterilization of inert surfaces and disinfection of living tissues. Our recent studies reveal that chemically active species such as reactive oxygen species generated by a low-power atmospheric pressure non- thermal plasma source obtained by ionizing helium gas mixed with ambient air are mandatory for their antimicrobial effects. In addition, they also initiate intracellular signaling pathways required for wound healing in eukaryotic cells. Thus, exposure to plasma sustains healing of tissue injuries both indirectly through microbicidal effects and directly by action on cells such as fibroblasts involved in tissue regeneration.

Bone morphogenetic proteins (BMPs) play a key role in bone and cartilage formation. For these properties, BMPs are employed in the field of tissue engineering to induce bone regeneration in damaged tissues. To overcome drawbacks due to the use of entire proteins, synthetic peptides derived from their parent BMPs have come out as promising molecules for biomaterial design. On the structural ground of the experimental BMP-2 receptor complexes reported in the literature, we designed three peptides, reproducing the BMP-2 region responsible for the binding to the type II receptor, ActRIIB. These peptides were characterized by NMR, and the structural features of the peptide-receptor binding interface were highlighted by docking experiments. Peptide-receptor binding affinities were analyzed by means of ELISA and surface plasmon resonance techniques. Furthermore, cellular assays were performed to assess their osteoinductive properties. A chimera peptide, obtained by combining the sequence portions 73-92 and 30-34 of BMP-2, shows the best affinity for ActRIIB in the series and represents a good starting point for the design of new compounds able to reproduce osteogenic properties of the parent BMP-2.

Adenosine A(2B) receptors regulate several physiological enteric functions. However, their role in the pathophysiology of intestinal dysmotility associated with inflammation has not been elucidated. Hence, we investigated the expression of A2B receptors in rat colon and their role in the control of cholinergic motility in the presence of bowel inflammation.Colitis was induced by 2,4-dinitrobenzenesulfonic acid (DNBS). Colonic A(2B) receptor expression and localization were examined by RT-PCR and immunofluorescence. The interaction between A(2B) receptors and adenosine deaminase was assayed by immunoprecipitation. The role of A(2B) receptors in the control of colonic motility was examined in functional experiments on longitudinal muscle preparations (LMPs).A(2B) receptor mRNA was present in colon from both normal and DNBS-treated rats but levels were increased in the latter. A(2B) receptors were predominantly located in the neuromuscular layer, but, in the presence of colitis, were increased mainly in longitudinal muscle. Functionally, the A(2B) receptor antagonist MRS 1754 enhanced both electrically-evoked and carbachol-induced cholinergic contractions in normal LMPs, but was less effective in inflamed tissues. The A(2B) receptor agonist NECA decreased colonic cholinergic motility, with increased efficacy in inflamed LMP. Immunoprecipitation and functional tests revealed a link between A(2B) receptors and adenosine deaminase, which colocalize in the neuromuscular compartment.Under normal conditions, endogenous adenosine modulates colonic motility via A2B receptors located in the neuromuscular compartment. In the presence of colitis, this inhibitory control is impaired due to a link between A2B receptors and adenosine deaminase, which catabolizes adenosine, thus preventing A(2B) receptor activation.

Introduction Nociceptive pain is one of the most common types of pain that originates from an injury involving nociceptors. Approximately 60% of the knee joint innervations are classified as nociceptive. The specific biological mechanism underlying the regulation of nociceptors is relevant for the treatment of symptoms affecting the knee joint. Intra-articular administration of exogenous hyaluronic acid (HA) in patients with osteoarthritis (OA) appears to be particularly effective in reducing pain and improving patient function. Methods We performed an in vitro study conducted in CHO cells that expressed a panel of opioid receptors and in primary rat dorsal root ganglion (DRG) neurons to determine if HA induces the activation of opioid peptide receptors (OPr) using both aequorin and the fluorescent dye Fura-2/AM. Results Selective agonists and antagonists for each OPr expressed on CHO cells were used to test the efficacy of our in vitro model followed by stimulation with HA. The results showed that HA induces stimulatory effects on the κ receptor (KOP). These effects of HA were also confirmed in rat DRG neurons, which express endogenously the OPr. Conclusions HA activates the KOP receptor in a concentration dependent manner, with a pEC50 value of 7.57.

Low temperature plasmas have been proposed in medicine as agents for tissue disinfection and have received increasing attention due to the frequency of bacterial resistance to antibiotics. Our previous studies (1) demonstrated that atmospheric-pressure cold plasma (APCP) generated by a new portable device that ionizes a flow of helium gas inactivated ocular pathogens in vitro. This study explored whether APCP inactivates ocular pathogens without causing significant tissue damage. We tested the APCP effects on cultured Pseudomonas aeruginosa, Staphylococcus aureus, A. fumigatus, ocular cells (conjunctival fibroblasts and keratocytes) and ex-vivo cornea. Exposure to APCP for 0.5–5 min significantly reduced microbial viability (colony-forming units) but not human cell viability (MTT assay and Tunel analysis). Since our previous study indicated that exposure to plasma increases intracellular reactive oxygen species (ROS) production, ROS levels in APCP exposed microorganisms and keratocytes were analyzed by 2’,7’-dichlorofluorescein diacetate (HDCF-DA) fluorescence. The potential genotoxic effects of plasma on cells and tissues were evaluated by analyses of thymine dimers (TD), genes and proteins involved in DNA damage and repair (OGG1, GPX, NRF2) at set time intervals. High levels of intracellular reactive oxygen species (ROS) were found in exposed microorganisms and cells. Immunoassay confirmed no induction of thymine dimers in corneal tissues. Conversely, a transient expression of genes and proteins recruited following oxidative stress was determined in ocular cells and corneas by qRT-PCR and Western blotting. In conclusion, a short application of APCP appears to be an efficient and rapid ocular disinfectant with ROS production likely causing pathogen killing and no substantial effects on ocular cells and tissues. The same APCP treatment to conjuntival fibroblasts and keratocytes caused a time-restricted formation of ROS and a change in some stress-response genes., Italian Journal of Anatomy and Embryology, Vol 116, No 1 (Supplement) 2011