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The article discusses the aesthetics and the politics of the so-called New Romanian Cinema (a major post-2005 sensation on the international art-cinema circuit), succintly assessing its novelty in both a national and an international cinematic context. It then proceeds to discuss Radu Jude’s debut feature, The Happiest Girl in the World (2009), as a NRC film, typical in some ways, atypical in others. The paper highlights the film’s critique of advertising and the nuclear family.

The paper parses the major existing literature, in English and Romanian, on the New Romanian Cinema (also called the Romanian New Wave) which became an international film-festival sensation in the mid-2000s. It maps the main lines of disagreement among the various scholars who have attempted to define this phenomenon. The paper argues that the Romanian cinema that had made its dazzling debut on the international scene some 15 years ago has gradually turned into something else – something harder to pin down to an aesthetic paradigm, to a core generational group of ten or so filmmakers, or to a limited set of ideological coordinates. Finally, the paper proposes that the evolving body of work of director Radu Jude (who has established himself in recent years as a major innovator) provides a good lens through which to take the measure of those aesthetic and political changes.

This article reviews a new Romanian translation of Rousseau’s Social contract. While attempting to grasp some of the challenges of translating Rousseau’s works, it also points towards the market-driven dynamics of the Romanian publishing landscape of translations from the classics of political thought.

"Jean Mabillon and Richard Simon were both eminent seventeenth-century scholars who practiced contextualizing critical philology in order to forge unbeatable scientifical instruments against the skeptics and reinforce the authority of historical documents. But Simon’s work produced a mutation of the meaning of authenticity that would prove subversive and would generate outrage. His sociological and institutionalist understanding of the history of sacred texts not only merged both their production and their transmission into a common, ontologically homogenous historical process, but also included a survey of the sociological and cultural circumstances that transformed the text into a real authority. Furthermore, this anti-essentialist understanding of the gradual formation of the text allowed a positive reevaluation of the tradition as a continuous practice aiming at keeping alive an already historical truth. Key words: historical criticism, biblical hermeneutics, Richard Simon, Jean Mabillon, Spinoza "

The aim of the present study was to investigate the potential antimicrobial activity of Olea europaea fatty oil against a collection of bacterial (Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa) and fungal (Candida albicans) clinical isolates. The antimicrobial and antibiofilm activity were evaluated by broth microdilution method for establishing the minimal inhibitory concentration (MIC) and microtiter assay for determining the minimal biofilm eradication concentration (MBEC). Some of the potential microbial targets of the fatty oil were investigated by flow cytometry (FCM). The results obtained hereby revealed that Olea europaea fatty oil inhibited microbial planktonic growth (MICs values of 5.23-41.8 mg/ mL) and biofilm development on inert substrata (MBEC values of 1.31-20.9 mg/ mL). The FCM measurements confirmed that the analyzed oil induced microbial membrane damages and inhibited microbial efflux pump activity.

In this review, we focus on gut microbiota profiles in infants and adults colonized (CDC) or infected (CDI) with Clostridioides difficile. After a short update on CDI epidemiology and pathology, we present the gut dysbiosis profiles associated with CDI in adults and infants, as well as the role of dysbiosis in C. difficile spores germination and multiplication. Both molecular and culturomic studies agree on a significant decrease of gut microbiota diversity and resilience in CDI, depletion of Firmicutes, Bacteroidetes, and Actinobacteria phyla and a high abundance of Proteobacteria, associated with low butyrogenic and high lactic acid-bacteria levels. In symptomatic cases, microbiota deviations are associated with high levels of inflammatory markers, such as calprotectin. In infants, colonization with Bifidobacteria that trigger a local anti-inflammatory response and abundance of Ruminococcus, together with lack of receptors for clostridial toxins and immunological factors (e.g., C. difficile toxins neutralizing antibodies) might explain the lack of clinical symptoms. Gut dysbiosis amelioration through administration of “biotics” or non-toxigenic C. difficile preparations and fecal microbiota transplantation proved to be very useful for the management of CDI.

Cellulases are a group of enzymes that are used in many biotechnological processes. Since most of the enzymes synthesised by mesophilic microorganisms are unstable in industrial environments, it is necessary to direct research towards extremophile cellulolytic microorganisms because the enzymes synthesised by them are stable and active even in harsh physicochemical conditions. In the present investigation, our aim was to isolate and identify some microbial cellulolytic strains from a hypersaline lake located in Romania and to determine their optimal growth conditions. Of a total of 25 microbial strains isolated, only one extreme halotolerant bacterial strain was able to produce an endoglucanase. Based on molecular identification, we identified this cellulolytic strain as a species of Bacillus genus, most closely related to Bacillus zhangzhouensis. Optimal growth conditions were found to be at 15°C, pH 7.5 and 2 M NaCl. Endoglucanase activity of this bacterial strain is influenced by both salinity and temperature. The most significant endoglucanase activity was detected in the presence of 3 M NaCl, after 72 h of incubation at 15°C. In this situation, the amount of glucose released from a volume of 0.5 mL of 2% (w/v) carboxymethyl cellulose substrate is equivalent to 2.05 mg. In conclusion, this study represents the first preliminary characterization of a B. zhangzhouensis strain that has the ability to degrade cellulose and that demonstrates tolerance to high salt concentrations.

El Patrimonio cultural material está expuesto a múltiples factores ambientales de riesgo que pueden afectar a su integridad y función cultural. Estos factores pueden ser físicos, químicos y también microbiológicos. El biodeterioro producido por hongos es un conocido causante de daño estético y estructural de los materiales, con un efecto mayor en el caso de deposición inadecuada o accidentes, como inundaciones o fugas de agua. Al mismo tiempo, la contaminación del aire con diferentes esporas de hongos puede constituir un factor de riesgo para la seguridad de los bienes patrimoniales. El rastreo de la biocontaminación aérea en museos tiene una doble utilidad: protección de la salud de los trabajadores en el museo ante bio-aerosoles peligrosos y el control de la presencia de agentes biodeteriorantes capaces de descomponer las piezas del museo. El presente documento presenta la metodología utilizada para la detección de especies de hongos en el aire de un museo etnográfico en Rumania, tanto en salas de almacenamiento, como de exposición. Los resultados preliminares muestran un predominio de géneros de hongos con actividad celulolítica reconocida, tales como Aspergillus, Penicillium y Cladosporium y una correlación directa entre los valores de temperatura y humedad con la concentración y los tipos de hongos presentes en el aire. Además, los medios de cultivo utilizados para el aislamiento de las especies fúngicas del aire se mostraron como un factor importante en la metodología de aislamiento.

This work presents a novel nano-modified coating for wound dressings and other medical devices with anti-infective properties, based on functionalized zinc oxide nanostructures and orange oil (ZnO@OO). The obtained nanosurfaces were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), selected area electron diffraction (SAED), differential thermal analysis-thermogravimetry (DTA-TG), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy. The obtained nanocomposite coatings exhibited an antimicrobial activity superior to bare ZnO nanoparticles (NPs) and to the control antibiotic against Staphylococcus aureus and Escherichia coli, as revealed by the lower minimal inhibitory concentration values. For the quantitative measurement of biofilm-embedded microbial cells, a culture-based, viable cell count method was used. The coated wound dressings proved to be more resistant to S. aureus microbial colonization and biofilm formation compared to the uncoated controls. These results, correlated with the good in vivo biodistribution open new directions for the design of nanostructured bioactive coating and surfaces, which can find applications in the medical field, for obtaining improved bioactive wound dressings and prosthetic devices, but also in food packaging and cosmetic industry.

Gentamicin is an aminoglycoside antibiotic with a wide spectrum of anti-bacterial activity, but however, due to its high solubility in water, it poorly penetrates inside the cells. This major inconvenient constitutes an important challenge for the treatment of intracellular bacterial infections, which might be solved using appropriate delivery systems for the targeted release of the bioactive agents at the intracellular sites of infection. Thus, in the case of antibiotics, the use drug delivery systems may contribute to increase their therapeutic activity against intracellular pathogens. This paper presents an efficient polymeric delivery system for the intracellular release of gentamicin based on bacterial polysaccharides.

In medicine, the surgical instruments are realized using a variety of stainless steel, such as curved elevators. The quality and the price of surgical instruments differ in function of the manufacturing processes and the material used. This paper, presents a prototype of elevator, manufactured using two types of additive manufacturing process Direct Metal Laser Sintering – DMLS process for beaks and stereolithography process for the handles. The handle was made by means of stereolithography with the printer PROJET 1500, using a plastic material D638. The design and the quality obtained for the prototype of elevator is better, thanks to the material used and to the manufacturing process. The prototype of handle manufactured by stereolithography using the UV rays presents very good mechanical resistance and allows to be sterilized. For the handle of the elevator prototype there was performed a FEM analysis to identify stress locations and displacements. In this paper, was realized a Attenuated Total Reflection (ATR) analysis on the plastic material D638 to be examined directly in the solid state without further preparation.

The purpose of this study was to investigate the response of HeLa cells to the interaction with inactivated Staphylococcus aureus cells and live challenge with herpes simplex virus (HSV).The results of this study are indicating that the interaction between the HeLa cells and S. aureus inactivated whole cells could modulate the host cell apoptosis and cytokine production, and therefore, influence the progression of HSV infection. The pre-treatment of HeLa cells with heat inactivated bacterial whole cells protects them from the occurrence of HSV mediated cytopathic effect, while the post viral infection treatment with bacterial cells prevents the high activation of bax/bcl-2 apoptotic pathway, a process that could change the fate of the infectious process triggered by the virus, and eventually reduce its multiplication rate. The pre-treatment of HeLa monolayer with inactivated bacterial cells 24 hours before the viral infection is increasing the expression level of TNF-a, IL-6 and IL-8 pro-inflammatory cytokines genes, also suggesting that bacterial antigens could contribute to the decrease of viral multiplication rate.

Chronic wounds represent an important burden on the healthcare system, requiring frequent hospitalizations and expensive treatments. It is now recognized that a primary factor contributing to a non-healing trajectory and a low therapeutic response is the biofilm infection. The purpose of this study was to identify the bacterial isolates collected from chronic skin wounds of hospitalized patients and to evaluate their antimicrobial susceptibility profiles, virulence factors, as well as the ability to develop biofilms in vitro. A number of 44 wound samples were collected from 39 patients. The isolated strains belonged to seven different microbial species, i.e.: Staphylococcus aureus (32 strains), followed by Pseudomonas aeruginosa (4), Escherichia coli (3), Klebsiella pneumoniae (2), Proteus mirabilis (1), Citrobacter freundii (1), group G β-hemolytic streptococci (1). In comparison to the other isolates, P. aeruginosa strains exhibited the highest capacity to develop complex biofilm structures in vitro, followed by S. aureus, with insignificant differences between MRSA and non-methicillin resistant isolates. The Enterobacteriaceae strains expressed less virulent phenotypes, lower adherence to epithelial cells and biofilm forming capacity, but also significant resistance phenotypes with a potential of unfavorable epidemiological outcome. The isolation of MRSA, ESBL-producing microorganisms and multiple antibiotic resistant P. aeruginosa suggests the potential risk of nosocomial spread and the potential severe outcome in case of bacteremia and sepsis. This study represents an important step in elucidating the host-wound microbiome interaction, by describing various resistance and virulence threats of microorganisms colonizing and/or infecting the chronic wounds. However, in order to establish a statistical relevant correlation, larger studies are needed.

In vitro antimicrobial and antioxidant activities of Amaranthus retroflexus leaves and inflorescence alcoholic (ethanol 70%) extracts of various concentrations ranging from 0.78 to 400 μL/ml were analyzed on different clinical and reference bacterial strains (Staphylococcus aureus, Bacillus subtills, Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii) and fungal strains (Candida albicans, C. famata, C. utilis, Saccharomyces cerevisiae) using agar disk diffusion method and broth dilution method (MIC determination) for antimicrobial activity and TEAC (Trolox capacity) assay for the evaluation of the antioxidant activity. The adapted diffusion method was used to test the antimicrobial effect of the extracts separately and in combination with a range of standard antibiotics, in order to evidence any synergic effects of A. retroflexus extracts on the antibiotics activity. The extracts showed the highest inhibitory effect against K. pneumoniae and B. subtilis with no activity against S. aureus among the bacterial strains, while in case of the fungal strains the most intensive effect was exhibited against C. famata by both extracts. The A. retroflexus leaves extract improved the ciprofloxacin and ticarcillin-clavulanic acid activity towards the P. aeruginosa clinical strain. The inflorescences extract significantly increased chloramphenicol activity on B. subtilis strain. The antioxidant activity assay showed that the studied extracts exhibited the ability to neutralize the free radicals leading to the conclusion that the tested extracts bear compounds with a broad spectrum of antimicrobial and antioxidant activity that could represent a potential alternative for treating various infectious diseases.

Intracellular invasion of professional phagocytic cells like monocytes and macrophages by a pathogen usually triggers the apoptosis of the host cells. The aim of this study was to evaluate if Pseudomonas aeruginosa, although not considered a classic intracellular pathogen, could adhere to endothelial cell surface, invade the intracellular compartment and subsequently induce apoptosis of the cells. The adherence and invasion capacity of P. aeruginosa to endothelial cells was monitored using Cravioto's adapted method. The apoptotic cells were evidenced by staining with Acridine orange/Ethidium bromide. The qualitative assay of bacterial adherence to the cellular substrate revealed that all tested strains adhered to endothelial cells surface, exhibiting a diffuse, aggregative or mixed (diffuse-aggregative or localized-aggregative) pattern and 20-70% adherence rates. The adherence of P. aeruginosa induced the reorganization of cytoskeleton filaments and formation of endocytic membrane expansions. Cell free P. aeruginosa culture supernatants did not induce any cell death response, as noticed in case of whole bacterial culture, showing the capacity to induce apoptosis of endothelial cells. The fluorescence microscopy examination revealed chromatin condensation, fragmented nuclei, and membrane blebbing and apoptotic bodies in pathogen invaded cells.

The purpose of this study was to determine the correlations between the concentration of free light chains (kappa, lambda and ratio kappa/lambda) and two other markers, M- protein and pathological total intact immunoglobulin in four groups of patients with intact immunoglobulin multiple myeloma (IIMM) at the diagnosis and during the treatment. In this study 354 samples coming from 46 patients with IIMM were assayed, out of which: 19, IgGkappa; 13, IgGlambda; 7, IgAkappa; 7, IgAlambda. At the diagnosis, immunofixation was positive in all samples and serum protein electrophoresis quantified M- protein for all patients. Free light chains concentrations were abnormal in 92.25% of patients with concentrations above the reference ranges in all patients with IgGkappa and IgAkappa MM. The intact immunoglobulins were elevated in 83.12% of cases. Pearson correlation coefficient showed correlations among the free light chains serum levels (kappa, lambda and ratio kappa/lambda), M- protein and intact immunoglobulins in two groups with IIMM (IgGlambda, IgAlambda). Spearman correlation coefficient values analysis showed that there is a good correlation between M-protein and FLCs (kappa, lambda and ratio kappa/lambda) in three patient groups (IgGkappa, IgGlambda and IgAlambda), excepting IgAkappa myeloma group where the correlation was insignificant. Regarding the intact immunoglobulin, Spearman coefficient showed significant correlations with FLCs concentrations in two groups (IgGlambda and IgAlambda) and an insignificant correlation in the group with IgGkappa MM. For the group of patients with IgAkappa myeloma, the Spearman coefficient showed that IgA concentrations did not correlate with the concentrations of FLCs. The individual correlation (for each patient) among FLCs, M- protein and intact immunoglobulins in 8 patients with IgGkappa IIMM proved to be more significant as compared with the degree of correlation established for the entire group of patients among these markers. Since the values obtained for the studied markers were not homogeneous, we have taken into account the Spearman coefficient values, which indicated the existence ofa correlation between the M-protein, FLCs and intact immunoglobulin.

Multidrug resistance and the increasing number of severe infections caused by Acinetobacter sp. strains are a major issue for intensive care units (ICUs), where patients with severe diseases and often destabilized physiological condition are admitted. The aim of this study was to investigate the antibiotic resistance profiles of 200 Acinetobacter spp. isolated from tracheal aspirates in patients admitted to ICU, Fundeni Clinical Institute (FCI).the samples were collected from intubated patients between January 2006-December 2007. The microbial strains were identified with the help of the BD Phoenix system. The investigation of the antibiotic resistance patterns was performed by agar disk diffusion method according to CLSI recommendations, and the production of metallo-betalactamases (MBL) was confirmed by MBL E-test.the majority of the studied strains (80%) were multidrug resistant with a high percentage of panresistance (32%). Metallo-beta-lactamases production among the strains with resistance to imipenem was high (over 83%), these strains being also resistant to the majority of the other tested antibiotics with the exception of colistin.Our results confirmed that the multidrug resistance is the major threat of Acinetobacter sp. infections, especially when they occur in high risk patients.

A series of N-(1-methyl-1 Hpyrazole-4-carbonyl)-thiourea derivatives were assessed for their in vitro antimicrobial and anti-pathogenic activity against twenty-two strains of Erwinia amylovora isolated from different regions in Romania. The compounds were solubilised in dimethylsulfoxide and screened for their in vitro antimicrobial activity. The qualitative screening of the susceptibility spectra of various strains to the compounds was performed by adapted diffusion techniques (distribution of the tested compound solution directly on the solid medium previously seeded with the bacterial inoculums). The quantitative assay of the minimal inhibitory concentration (MIC, microg/mL) was based on liquid medium two-fold microdilutions. The subinhibitory concentrations of the tested substances were investigated for their influence on biofilm development on inert substrata. The present study showed that six new thiourea compounds exhibited a low antibacterial activity (MIC values500 microg/ml), but the subinhibitory concentrations inhibited the biofilm development on inert substrata. Thus, these results could suggest the usefulness of the tested compounds as control agents for preventing the first stage (colonization) of the infection with the fire blight pathogen.

Monoclonal gammopathies are a group of disorders, referred to as paraproteinaemias, dysproteinaemias or immunoglobulinopathies, associated with monoclonal proliferation of plasma cells. Monoclonal immunoglobulin secreted by these cells is an indicator of clonal proliferation. The aim of this study is to analyze the efficiency of three methods: serum protein electrophoresis (SPE), serum protein immunofixation (IFE) and FLC (free light chain) assay for the diagnosis and monitoring of the tumor burden in multiple myeloma. In this study we have presented the dynamic evolution of 7 patients with intact immunoglobulin multiple myeloma (IIMM) (2 IgG, kapa; 3 IgG, lambda; 1 IgA, kappa; 1 IgA, lambda) and 2 patients with light chain multiple myeloma before and after autologous peripheral blood stem cell transplantation (PBSCT). All 7 patients fulfilled the four criteria for the diagnosis of IIMM: bone marrow plasma cells exceeding 20%, lytic bone lesions, identification and quantification of M protein by scanning densitometry of electrophoresis gels, IFE (immunofixation protein electrophoresis) confirmed and typed the M protein. All patients had been given cytotoxic chemotherapy (VAD or VELCADE) before autologous (PBSCT). In two of the patients with IIMM both SPE and kappa/lambda ratio fell towards normal range after autologous PBSC and both reported a relapse of the disease after 23 months and 19 months respectively. SPE could not normalize after chemotherapy and transplantation in three patients with IIMM, the kappa/lambda ratio being the only marker used to monitor the tumor kill. In one patient the kappa/lambda ratio could not normalize even after PBSCT still indicating the presence of plasma cell disorder at the time when IFE was still negative. 16 months after PBSCT both SPE and FLC indicated a relapse of the disease. Classical SPE failed to demonstrate the presence of M-protein in light chain multiple myeloma, the diagnosis being established by using IFE and the FLC assay. Because IFE is a qualitative method and its interpretation may be sometimes subjective, FLC was the only method used to follow the disease course. The measurement of kappa/lambda ratio proved to be more sensitive than SPE, IFE and the levels of free light chains kappa or lambda individually indicating whether the treatment is effective or not.

The opportunistic pathogen Pseudomonas (Ps.) aeruginosa causes severe infections, particularly in immunocompromised individuals and patients with cystic fibrosis (CF). A serious side effect of antibiotic therapy in Ps. aeruginosa infections is the development of resistance to antibiotics. During the infection process Ps. aeruginosa forms biofilms, rendering bacterial cells more resistant to disinfectants, antibiotics and the action of host immune defense effectors. Pseudomonas aeruginosa employs the intercellular communication system, known as quorum sensing (QS) to coordinate the expression of tissue-damaging factors. Since the QS systems controls the production of different virulence factors, it is possible that the inhibition of its regulatory activity to severely compromise the ability of Ps. aeruginosa to cause infections in humans. Many studies have shown that some probiotic strains exhibit inhibitory activity on different virulence properties of pathogenic bacteria (adherence to cellular or inert substrate, soluble virulence factors expression). The aim of the present study was to investigate by real-time RT-qPCR the influence of probiotic culture soluble factors on the QS genes expression in 30 Ps. aeruginosa strains isolated from patients hospitalized in the National Institute for Cardiovascular Infections, Prof. C.C. Iliescu Fundeni Hospital, Bucharest. The results of the real time RT-qPCR have shown that in all Ps. aeruginosa strains grown in the presence of probiotic culture sterile filtrates, the level of QS genes expression was reduced comparatively with those from control cultures. In conclusion, these results proved that the inhibition of virulence factors regulation mechanisms by soluble molecules secreted by probiotics could represent an interesting way pathogenicity and virulence attenuation in Ps. aeruginosa nosocomial strains.

Considering the important role of cytokines in the initiation and evolution of the inflammatory process induced by Shigella and EIEC strains, the purpose of this study was the characterization of the secretory patterns of HeLa cells induced by Shigella ssp. and EIEC strains and to link the obtained results with the invasiveness level of bacterial strains on this cellular line. During this study there were analyzed two EIEC strains and 12 strains of the following Shigella species: 2 Sh. flexneri 2a, 2 Sh. flexneri 3a, 2 Sh. flexneri 4a, 2 Sh. boydii, 2 Sh. sonnei strains isolated in Romania during 2005 from children with dysentery and diarrhoea and confirmed for their invasive ability by Sereny test. The level of the main pro and anti-inflammatory cytokines, IL-1 beta, IL-6, IL-10, IL-13, IL-17 and TNF-alpha induced by whole bacterial cultures as well as by their soluble mediators was determined by an ELISA test. Our results showed that HeLa cells can be used not only for the qualitative and quantitative assessment of Shigella and EIEC strains invasion ability, but also as a simple work procedure for the investigation of an in vitro complex crosstalk communication mechanisms that involves physical interactions between bacterial cells and epithelial cells (adhesins and complementary receptors) and pro- and anti-inflammatory molecules regulation.The majority of the analyzed Shigella serogroups, with the exception of Shigella sonnei and EIEC strains, inhibited the inflammatory response by reducing the expression of majority of pro-inflammatory cytokines, i.e., IL-1 beta, IL-6, TNF-alpha and IL-17. The reduced cost of the in vitro procedure, the possibility of results interpretation and the strict regulations concerning the use of animals for experimental purposes are the main reasons that support the implementation of such an in vitro test in the research labs.

The aim of present study was to investigate the microbial colonization of worn contact lenses (CLs) and to evaluate the inhibitory effect of pomelo (Citrus maxima) peels essential oil on the biofilm development on unworn CLs. The essential oil was isolated by steam distillation and analyzed by gas chromatography coupled with mass spectrometry, twenty compounds being isolated. The antimicrobial activity of pomelo oil was tested against S. epidermidis and P. aeruginosa strains, known for their ability to develop biofilms on prosthetic devices, by qualitative screening methods and quantitative assay of the minimal inhibitory concentrations (MIC) in order to evaluate the antibiofilm activity. Our study revealed that all worn CLs where 100% colonized by staphylococci and Enterobacteriaceae strains. The pomelo essential oil inhibited the development of bacterial biofilms formed by Gram-positive and Gram-negative microorganisms on soft CLs, its antibiofilm activity being specific and dependent on different physical parameters (contact time and temperature). The architecture of bacterial biofilms developed on soft contact lenses was analyzed using confocal scanning laser microscopy (CSLM).

Recent public announcements stated that 60% to 85% of all microbial infections involve biofilms developed on natural tissues (skin, mucosa, endothelial epithelia, teeth, bones) or artificial devices (central venous, peritoneal and urinary catheters, dental materials, cardiac valves, intrauterine contraceptive devices, contact lenses, different types of implants). Prosthetic medical devices are risk factors of chronic infections in developed countries and these infections are characterized by slow onset, middle intensity symptoms, chronic evolution and resistance to antibiotic treatment. In case of biofilm development, a series of genes (40-60% of the prokaryotic genome) are modulated (activated/inhibited) by complex cell to cell signalling mechanisms and the biofilm cells become phenotypically distinct from their counterpart--free cells, being more resistant to stress conditions (including all types of antimicrobial substances); this resistance is phenotypical, behavioural and, more recently, called TOLERANCE. Four major mechanisms can account for biofilm antibiotic tolerance: (1) the failure of antibiotic penetration into the depth of a mature biofilm due to the biofilm matrix; (2) the accumulation of high levels of antibiotic degrading enzymes; (3) in the depth of biofilm, cells are experiencing nutrient limitation entering in a slow-growing or starved state; slow-growing or non-growing cells being not highly susceptible to antimicrobial agents, this phenomenon could be amplified by the presence of phenotypic variants or "persisters" and (4) biofilm's bacteria can turn on stress-response genes and switch to more tolerant phenotypes on exposure to environmental stresses; (5) genetic changes, probably selected by different stress conditions, such as mutations and gene transfer could occur inside the biofilm. In these conditions, biofilm associated infections require a different approach, both clinically and paraclinically.

Vibrio (V.) parahaemolyticus is an aquatic halophilic bacteria which produces gastroenteritis and in rare cases septicaemia after the consumption of raw or under-cooked contaminated seafood.The severity of diarrheal illness caused by this bacterium is closely related to the presence of two types of hemolysins (the thermostable direct hemolysin-TDH and TDH related hemolysin-TRH) and also of type III secretion system (TTSS) proteins. The TTSS type 1 induces a wide array of effects on infected HeLa cells such as autophagy, oncosis, cell rounding and lysis. Previous studies have shown that heat shock proteins have the ability to stimulate the production of interleukins in different cellular cultures. In our studies we have stimulated two cellular lines (HeLa and human diploid cells) with different V. parahaemolyticus culture fractions in order to observe the effect on cytokines production. Thus, the purpose of this study was to analyze the expression of IL-1, IL-2, IL-4, IL-6, IL-10 and TNF-alpha induced by the cell treatment with total cellular lysate, periplasmic fractions and culture supernatants extracted from V. parahaemolyticus exposed to normal and also to stress conditions. The ELISA assay of the cytokine profile of the HeLa and HDC cell lines stimulated with different bacterial fractions revealed that in the V. parahemolyticus cultures submitted to osmotic and heat shock stress are accumulating factors (probably heat shock proteins) which are exhibiting immunomodulatory activity, responsible for the induction of a pro-inflammatory response associated with increased levels of IL-6 and TNF-alpha expression, however balanced by the stimulation of the anti-inflammatory cytokine IL-4 synthesis.