Roumanian Archives Of Microbiology And Immunology[JOURNAL]
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200 papers
Implant microparticles--a new concept for non-invasive cancer therapy.
Surdu-Bob CC, Coman C, Călugăru A … +3 more , Cremer L, Bădulescu M, Vlase E
Roum Arch Microbiol Immunol · 2011 · PMID 21717810
Some progress in cancer research was possible in recent years mainly due to important advances in nanotechnology. However, clinical use of nanomaterials is still hindered by limitations. In search of better performance a...
Some progress in cancer research was possible in recent years mainly due to important advances in nanotechnology. However, clinical use of nanomaterials is still hindered by limitations. In search of better performance and control of inoculated materials, the efficiency and toxicity of SBBC implant particles was assessed. B16 tumoral cells (murine melanoma) were subjected to SBCC particles using in vitro and in vivo experimental models. In vitro experiments concerning the growth inhibition of tumoral cells using SBCC particles were performed by Flow Cytometry and by MTT Assay. In vivo experimental model (C57BL/6 mice) was used to complete this investigation: weight, viability and tumoral dimension were monitored. An anti-proliferative activity on B16 tumoral cells and an ability to produce apoptosis were observed. A reduction of tumoral volume and a 54% survival rate in the treated animals compared to the controls was obtained. Our preliminary results showed that the SBCC implants were effective against B16 melanoma cells, while there is no toxicity associated.
Bacterial extract cantastim activates macrophages via TLR-2.
Caraş I, Tucureanu C, Pitica R … +1 more , Sălăgeanu A
Roum Arch Microbiol Immunol · 2011 · PMID 21717809
CANTASTIM is a second generation bacterial immunomodulator. The aim of this study was to examine the mechanism by which bacterial immunomodulator CANTASTIM induces production of inflammatory cytokines in monocytes/macrop...
CANTASTIM is a second generation bacterial immunomodulator. The aim of this study was to examine the mechanism by which bacterial immunomodulator CANTASTIM induces production of inflammatory cytokines in monocytes/macrophages. Proinflammatory cytokines were induced in PMA-differentiated THP-1 cells by stimulation with TLR agonists and CANTASTIM in the presence or absence of anti-TLR blocking antibodies or isotype matched control antibodies. Also, RNA interference was used to knockdown TLR2 or TLR4 expression in PMA-differentiated THP-1 cells before stimulation. As expected, induction of TNF-alpha and IL-6 by TLR4 agonist LPS was inhibited in a significant manner by anti-TLR4 but not by anti-TLR2 antibody. Unexpectedly, treatment with anti-LR2 blocking antibody inhibited only IL-6 production induced by Pam3CSK4 while the level of TNF-alpha was unchanged. When cells were stimulated by TLR2 agonist heat-killed Listeria monocytogenes the release of TNF-alpha was significantly attenuated by anti-TLR2 antibodies. Silencing of TLR2 led to a statistically significant inhibition of TNF-alpha secretion induced by TLR2 agonist while siRNA silencing of TLR4 did not affect the response to TLR2 agonist. Cells exposed to CANTASTIM produced significant levels of pro-inflammatory cytokines but the levels were lower than LPS-stimulated cells. Production of both cytokines was inhibited by treatment with anti-TLR2 blocking antibody and not by anti-TLR4 antibody. Silencing of TLR2 led to a statistically significant inhibition of TNF-a secretion induced by CANTASTIM while silencing of TLR4 had no effect on the response to CANTASTIM. These results support the hypothesis that CANTASTIM may exert its immunomodulatory and adjuvant activities through interaction of its bacterial components with TLR2.
Immunodetection of added glycomacropeptide in milk formulas and milk powders.
Oancea S, Stoia M
Roum Arch Microbiol Immunol · 2011 · PMID 21717808
The present study aimed the detection of fraudulent manipulation of milk powder with a low cost component--whey powder, by applying the immunochromatographic assay to identify glycomacropeptide. Five commercial milk powd...
The present study aimed the detection of fraudulent manipulation of milk powder with a low cost component--whey powder, by applying the immunochromatographic assay to identify glycomacropeptide. Five commercial milk powder samples of various brands from the national market were analyzed: lactose enriched milk powder type 26, two whole milk powders, vitamin enriched milk powder and full cream milk powder. Our results showed additional whey (1-2%) in 60% of the selected samples after casein removal by precipitation with 20% trichloracetic acid. Another investigated sample--the enriched UHT milk for children aged 4-12 years--proved addition of whey. Other two commercial toddler formula milk powder samples of different brands were used for comparison for the presence of glycomacropeptide. The first sample which was regularly labeled as containing whey protein concentrate was found positive for glycomacropeptide in accordance with the label information, while the second one not containing whey proteins as specified by the product label, was found negative for glycomacropeptide, these two samples being in accordance with the actual legislation.
Assessing the efficiency of free light chain assay in monitoring patients with multiple myeloma before and after autologous stem cell transplantation along with serum protein electrophoresis and serum protein immunofixation.
Dogaru M, Lazăr V, Coriu D
Roum Arch Microbiol Immunol · 2011 · PMID 21717807
Monoclonal gammopathies are a group of disorders, referred to as paraproteinaemias, dysproteinaemias or immunoglobulinopathies, associated with monoclonal proliferation of plasma cells. Monoclonal immunoglobulin secreted...
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.
In vitro antimicrobial activity of Romanian medicinal plants hydroalcoholic extracts on planktonic and adhered cells.
Stanciuc AM, Gaspar A, Moldovan L … +3 more , Saviuc C, Popa M, Măruţescu L
Roum Arch Microbiol Immunol · 2011 · PMID 21717806
The aim of this study was to assess the antibacterial and antifungal potential of some Romanian medicinal plants, arnica--Arnica montana, wormwood--Artemisia absinthium and nettle--Urtica dioica. In order to perform this...
The aim of this study was to assess the antibacterial and antifungal potential of some Romanian medicinal plants, arnica--Arnica montana, wormwood--Artemisia absinthium and nettle--Urtica dioica. In order to perform this antimicrobial screening, we obtained the vegetal extracts and we tested them on a series of Gram-positive and Gram-negative bacteria, and also against two fungal strains. The vegetal extracts showed antimicrobial activity preferentially directed against the planktonic fungal and bacterial growth, while the effect against biofilm formation and development was demonstrated only against S. aureus and C. albicans. Our in vitro assays indicate that the studied plant extracts are a significant source of natural alternatives to antimicrobial therapy, thus avoiding antibiotic therapy, the use of which has become excessive in recent years.
Mitochondrial DNA mutations in patients with HRHPV-related cervical lesions.
Goia-Ruşanu CD, Iancu IV, Botezatu A … +4 more , Socolov D, Huică I, Pleşa A, Anton G
Roum Arch Microbiol Immunol · 2011 · PMID 21717805
High risk human papillomaviruses (hr-HPV) are known to be the etiological agents of cervical cancer disease. On the other hand, other cofactors are considered to be important in cervix carcinogenesis. Mutations in mitoch...
High risk human papillomaviruses (hr-HPV) are known to be the etiological agents of cervical cancer disease. On the other hand, other cofactors are considered to be important in cervix carcinogenesis. Mutations in mitochondrial DNA (mtDNA) as well as alterations in mtDNA content have been reported in numerous cancers examined to date. The D-loop region has been shown to be a mutational "hot spot" in human cancer. In order to evaluate the role of mtDNA mutations in cervical lesions progression, cervical specimens (from 79 women, 29-65 years old) were investigated. DNA was isolated (High Pure PCR Template, Roche Diagnostics) from cervical cells from patients with different cytology (normal cervical epithelium, ASCUS-Atypical Squamous Cells of Undetermined Significance, LGSIL-Low-Grade Intraepithelial Lesion, HGSIL-High-Grade Intraepithelial Lesion and SCC-Squamous Cell Carcinoma) and tested for HPV DNA presence (Linear Array HPV Genotyping Test, Roche Diagnostics). To elucidate a causative role of mtDNA in cervical lesions, mtDNA mutations were investigated using Mutector mtDNA kit (TrimGen Corporation). In patients with normal and ASCUS cytology, mtDNA mutations were absent. 16.66% of LGSIL patients presented mutations in D-loop region whereas 28.57% HGSIL cases showed mutations in mtDNA. Mutations were detected in 66.66% cases of SCC cases. These studies provide strong evidence that instability in the D-loop region of mtDNA may be involved in cervical dysplasia. We suggested that mtDNA mutations may play a role in cervical precursor lesions and cancer but their role in the mechanism of carcinogenesis remains to be solved.
Antimicrobial activity of plant essential oils against bacterial and fungal species involved in food poisoning and/or food decay.
Lixandru BE, Drăcea NO, Dragomirescu CC … +6 more , Drăgulescu EC, Coldea IL, Anton L, Dobre E, Rovinaru C, Codiţă I
Roum Arch Microbiol Immunol · 2010 · PMID 21462837
The currative properties of aromatic and medicinal plants have been recognized since ancient times and, more recently, the antimicrobial activity of plant essential oils has been used in several applications, including f...
The currative properties of aromatic and medicinal plants have been recognized since ancient times and, more recently, the antimicrobial activity of plant essential oils has been used in several applications, including food preservation. The purpose of this study was to create directly comparable, quantitative data on the antimicrobial activity of some plant essential oils prepared in the National Institute of Research-Development for Chemistry and Petrochemistry, Bucharest to be used for the further development of food packaging technology, based on their antibacterial and antifungal activity. The essential oils extracted from thyme (Thymus vulgaris L.), basil (Ocimum basilicum L.), coriander (Coriandrum sativum L.), rosemary (Rosmarinus officinalis L.), sage (Salvia officinalis L.), fennel (Foeniculum vulgare L.), spearmint (Mentha spicata L.) and carraway (Carum carvi L.) were investigated for their antimicrobial activity against eleven different bacterial and three fungal strains belonging to species reported to be involved in food poisoning and/or food decay: S. aureus ATCC 25923, S. aureus ATCC 6538, S. aureus ATCC 25913, E. coli ATCC 25922, E. coli ATCC 35218, Salmonella enterica serovar Enteritidis Cantacuzino Institute Culture Collection (CICC) 10878, Listeria monocytogenes ATCC 19112, Bacillus cereus CIP 5127, Bacillus cereus ATCC 11778, Candida albicans ATCC 10231, Aspergillus niger ATCC 16404, Penicillium spp. CICC 251 and two E. coli and Salmonella enterica serovar Enteritidis clinical isolates. The majority of the tested essential oils exibited considerable inhibitory capacity against all the organisms tested, as supported by growth inhibition zone diameters, MICs and MBC's. Thyme, coriander and basil oils proved the best antibacterial activity, while thyme and spearmint oils better inhibited the fungal species.
Quantitative real-time PCR study of the influence of probiotic culture soluble fraction on the expression of Pseudomonas aeruginosa quorum sensing genes.
Cotar AI, Chifiriuc MC, Dinu S … +3 more , Pelinescu D, Banu O, Lazăr V
Roum Arch Microbiol Immunol · 2010 · PMID 21462836
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. a...
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.
Antimicrobial activity of copper and silver nanofilms on nosocomial bacterial species.
Codiţă I, Caplan DM, Drăgulescu EC … +5 more , Lixandru BE, Coldea IL, Dragomirescu CC, Surdu-Bob C, Bădulescu M
Roum Arch Microbiol Immunol · 2010 · PMID 21462835
Contaminated surfaces are possible vehicles in infection transmission. It is known that both Copper (Cu) and Silver (Ag) efficiently inactivate microbes by direct contact. Aiming at using these metals for benefitting fro...
Contaminated surfaces are possible vehicles in infection transmission. It is known that both Copper (Cu) and Silver (Ag) efficiently inactivate microbes by direct contact. Aiming at using these metals for benefitting from their antimicrobial effect, but to avoid subsequent toxic effects, we evaluated the antimicrobial activity of nanometric thin Silver and Copper films covering less expensive materials. Using a modified version of the Japan Industrial Standard JIS Z 2801:2000, we demonstrated the antimicrobial activity of the surfaces covered with metal ions nanofilms on microorganisms possibly involved in nosocomial infections and on Bacillus anthracis, bacteria with possible implication in bioterrorist attacks. Copper covered surfaces proved to have better antimicrobial activity than Silver surfaces. Silver covered surfaces showed better activity on Gram negative bacteria than on Gram positive cocci. Going deeper with studies on antimicrobial effects using new methods with better direct and/or functional discriminatory capacity is needed in order to provide additional information on the mechanisms of Silver and Copper nanofilms antimicrobial activity.
A multiple typing approach is recommended for the laboratory-based surveillance of salmonellosis in Romania.
Usein CR, Străuţ M, Palade AM … +5 more , Tatu-Chiţoiu D, Ciontea SA, Băltoiu MC, Dinu S, Damian M
Roum Arch Microbiol Immunol · 2010 · PMID 21462834
In Romania, Salmonella enterica serovar Typhimurium isolates are currently typed by antimicrobial resistance profiles and phage typing, as part of the national laboratory-based surveillance system of human enteric infect...
In Romania, Salmonella enterica serovar Typhimurium isolates are currently typed by antimicrobial resistance profiles and phage typing, as part of the national laboratory-based surveillance system of human enteric infections. The aim of the present study was to assess the added value of complementing this approach with molecular fingerprinting, namely pulsed-field gel electrophoresis (PFGE) and multiple-locus variable-number tandem-repeats analysis (MLVA). Thirty-six S. Typhimurium isolates received by the Reference Center for Human Salmonella Infections for confirmation and typing from the Microbiology Departments of three Public Health Authorities, were selected for this study. Phage typing revealed that 14 isolates (39%) were nontypeable (NT). Twenty-two isolates were assigned to 5 phage types: DT193 (11 isolates), U302 (7 isolates), DT116 (2 isolates), DT41 (1 isolate) and DT86 (1 isolate). Antimicrobial susceptibility testing showed that all the NT and DT116 isolates were multidrug resistant and extended-spectrum betalactamase producers. All the examined isolates were typeable when using the molecular approach. Both methods gave conclusive and comparable results, documenting the genetic relatedness and discriminating the outbreak isolates from sporadic cases. We conclude that in order to improve outbreak investigation and surveillance of salmonellosis in Romania, the current routine typing of Salmonella isolates should be complemented with at least one of these DNA fingerprinting methods.
Plasticity of regulatory T cells under cytokine pressure.
Diaconu CC, Neagu AI, Lungu R … +7 more , Tardei G, Alexiu I, Bleotu C, Economescu MC, Bumbăcea RS, Pele I, Bumbăcea D
Roum Arch Microbiol Immunol · 2010 · PMID 21462833
CD4+ T helper (Th) cells have been divided into different subsets as defined by their cytokine products and functions after their activation. CD4+ T cell subsets are continuously discovered and until now Th1, Th2, Th9, T...
CD4+ T helper (Th) cells have been divided into different subsets as defined by their cytokine products and functions after their activation. CD4+ T cell subsets are continuously discovered and until now Th1, Th2, Th9, Th17, and regulatory T (Treg) cells have been almost unanimously recognized but yet not completely characterized. The selective production of cytokines by each of the subsets is probably the master key of the mechanisms of immune regulation. The cytokine milieu is extremely important on deciding the fate of T cells. Generally, more than one cytokine is needed for differentiating to a particular lineage and just recently it was shown that this status quo of commitment could be challenged. It is well known that cytokines bind to Type I/II cytokine receptors signaling via Janus kinases (JAKs) followed by activation of Signal Transducer and Activator of Transcription (STAT). STAT molecules work together with other transcription factors (Foxp3, RORgammat and RORalpha, T-bet, GATA3, Runx 1, NFAT, etc.) also controlled by cytokines, in modulating the Th phenotype and functions. In this review, we analyze the plasticity of Treg population focusing on the most recent discoveries on how microenvironmental cytokines refine/modify Treg phenotype and function, thus changing their fate.
Proinflammatory cytokines: an insight into pancreatic oncogenesis.
Miron N, Miron MM, Milea VG … +1 more , Cristea V
Roum Arch Microbiol Immunol · 2010 · PMID 21462832
Pancreatic cancer is a highly lethal disease, being one of the five leading death causes among oncologic patients. It is usually diagnosed late due to the paucity of clinical signs, and the current therapy means have lim...
Pancreatic cancer is a highly lethal disease, being one of the five leading death causes among oncologic patients. It is usually diagnosed late due to the paucity of clinical signs, and the current therapy means have limited success. One of the documented risk factors for developing pancreatic adenocarcinoma is chronic pancreatitis. It is postulated that a chronic inflammatory disease has a potential of evolving toward neoplasia, a fact that could account for a percentage of the pancreatic cancers. Starting from this assumption, we intended to analyze the serum reflection of some molecules with proinflammatory roles, and compare them in healthy individuals, in patients with chronic pancreatitis and with pancreatic adenocarcinoma. Additionally, we performed a biochemical and hematological assessment of the study groups, and compared the results with the immunological parameters analyzed in the same subjects. We found significantly higher levels of Tumor Necrosis Factor-alpha and Interleukin 6 in chronic pancreatitis and pancreatic adenocarcinoma sera (with higher levels in the pancreatitis group than in the cancer group), compared to healthy controls. Additionally, we found significantly higher levels of interleukin 8 and Macrophage Inflammatory Protein-3 alpha in pancreatic cancer, compared to chronic pancreatitis and controls. We also identified numerous correlations between the abovementioned cytokines/chemokines and biochemical parameters, not very much studied before. Our results plead for a pathogenic role of chronic inflammation in pancreatic carcinogenesis, thus offering a potential tool for earliy diagnose or targets for therapy.
IgG-F-actin antibodies in celiac disease and dermatitis herpetiformis.
Samaşca G, Băican A, Pop T … +5 more , Pîrvan A, Miu N, Andreica M, Cristea V, Dejica D
Roum Arch Microbiol Immunol · 2010 · PMID 21462831
Anti-actin antibodies are found in 52-85% of patients with autoimmune hepatitis or chronic active hepatitis and in 22% of patients with primary biliary cirrhosis. In patients with celiac disease, anti-actin antibodies co...
Anti-actin antibodies are found in 52-85% of patients with autoimmune hepatitis or chronic active hepatitis and in 22% of patients with primary biliary cirrhosis. In patients with celiac disease, anti-actin antibodies correlate with the degree of villous atrophy. Studies on their involvement in celiac disease and dermatitis herpetiformis in Romania have not been done. The purpose of this study was to evaluate of the quality of IgG anti-F-actin antibodies (IgG-AAA) tests compared with IgA tissue transglutaminase antibodies (IgA-TgA) having IgA endomysial antibody (IgA-EmA) as gold standard in celiac disease and dermatitis herpetiformis and to see if there is any relationship between them. The study included 70 pediatric patients with celiac disease under gluten-free diets and 10 adult patients with dermatitis herpetiformis, during 2010. The IgG-AAA antibodies levels were determined by ELISA. Assessing the qualities of IgG-AAA compared to IgA-TgA, we obtained the following values sensitivity (Se) 27.8%, specificity (Sp) 79.4%, respectively Se 88.9%, Sp 79.4% in celiac disease and Se 33.3%, Sp 100%, respectively Se 100%, Sp 100% in dermatitis herpetiformis. Also, there was a prevalence of 24.3% and 30% of IgG-AAA in the two groups of patients, but no statistically significant associations were found. Therefore, we concluded that IgG-AAA can not replace IgA-TgA in children patients with celiac disease under gluten-free diets and in adult patients with dermatitis herpetiformis. AAA-IgG serum activity in both diseases exist, but without a relationship of association with them.
Cytokine profiles of HeLa and human diploid cells induced by different fractions of Vibrio parahaemolyticus cultures exposed to stress conditions.
Chifiriuc MC, Bleotu C, Pîrcălăbioru G … +5 more , Israil AM, Dinu S, Rută SM, Grancea C, Lazăr V
Roum Arch Microbiol Immunol · 2010 · PMID 21434593
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 illn...
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.
Tracking down of laryngo-pharyngeal metastasis.
Bleotu C, Popescu CR, Anton G … +8 more , Pleşa A, Grigore R, Welt L, Aldea I, Draguşel R, Chivu M, Matei G, Cernescu C
Roum Arch Microbiol Immunol · 2010 · PMID 21434592
BACKGROUND: In Romania, the most optimistic statistics give a 5 years survival rate in approximately 33% of laryngo-pharyngeal cancer patients. Considering that a cell carrying the viral DNA is originating from primary t...
BACKGROUND: In Romania, the most optimistic statistics give a 5 years survival rate in approximately 33% of laryngo-pharyngeal cancer patients. Considering that a cell carrying the viral DNA is originating from primary tumor, we have tested whether HPV DNA could be detected in the blood cell of patients with laryngeal cancer as a marker of disease progression and metastases. METHODS: The study was performed on 85 patients (59 +/- 8.7 age) with laryngo-pharyngeal cancer. HPV DNA was detected in tumor using nested PCR with consensus primers, and also in local lymph nodes and/or blood cells from patients HPV positive in primary tumor. RESULTS: HPV DNA was detected in 75.29% of analyzed tumours, and all HPV16 positive samples were confirmed by mRNA E6 expression. 56.3% of patients presented HPV DNA in peripheral circulation as confirmed by PCR with E6 HPV16 specific primers followed by Southern Blot. CONCLUSION: Our results sustain that the detection of HPV DNA in blood is a "surrogate marker" of metastasis when extension of metastasis cannot be estimated, this observation is very important for management of cancer patients with laryngopharyngeal localization.
The inhibitory activity of pomelo essential oil on the bacterial biofilms development on soft contact lenses.
Saviuc C, Dascălu L, Chifiriuc MC … +6 more , Rădulescu V, Oprea E, Popa M, Hristu R, Stanciu G, Lazăr V
Roum Arch Microbiol Immunol · 2010 · PMID 21434591
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...
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).
The modulation of hela cells secretory patterns by invasive Shigella spp. and enteroinvasive E. coli bacterial cells and their soluble components.
Chifiriuc MC, Bleotu C, Măruţescu L … +2 more , Cristea D, Lazăr V
Roum Arch Microbiol Immunol · 2010 · PMID 21434590
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 o...
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.
Medical significance and new therapeutical strategies for biofilm associated infections.
Lazăr V, Chifiriuc MC
Roum Arch Microbiol Immunol · 2010 · PMID 21434589
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, peri...
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.
Controversies in the laboratory diagnosis of celiac disease in children; new haplotypes discovered.
Samaşca G, Iancu M, Butnariu A … +5 more , Pîrvan A, Andreica M, Miu N, Constantinescu I, Dejica D
Roum Arch Microbiol Immunol · 2010 · PMID 21434588
The latest consensus on celiac disease in 2008, under the auspices of the International Societies of Pediatric Gastroenterology, Hepatology and Nutrition, shows that HLA DQ2/DQ8 typing indicates the highest negative pred...
The latest consensus on celiac disease in 2008, under the auspices of the International Societies of Pediatric Gastroenterology, Hepatology and Nutrition, shows that HLA DQ2/DQ8 typing indicates the highest negative predictive value for celiac disease, which would exclude the diagnosis of celiac disease. In Romania, there are no studies on the implication of HLA-DQ2/DQ8 in celiac disease in children. The aim of our study was to analyze the significance of genetic tests, with a focus on negative HLA-DQ2/DQ8 cases, as well as to determine the main haplotypes involved in celiac disease in children. We tested in 37 children with old celiac disease, confirmed based on the presence of intestinal villi changes on duodenal biopsy, the IgA anti-tissue transglutaminase antibodies (TgA-IgA) by ELISA and the IgA anti-endomysium antibodies (EmA-IgA) by indirect immunofluorescence, compared to HLA-DQ2/DQ8 typing by polymerase chain reaction (PCR). In 25 children, the determined HLA haplotypes predominantly belonged to DQ2, and in 3 children we report the presence of a new haplotype, DR3-DQ2/DR4-DQ8, formed by pattern 1, DR3-DQ2-the DQA1*0501 and DQB1*0201 alleles, and pattern 5, DR4-DQ8-the DQA1*0301 and DQB1*0302 alleles. In 9 children, genetic tests were negative for celiac disease. The identification of HLA-DQ2/DQ8 provides additional data in the diagnosis of celiac disease, but a rigid algorithm in the diagnosis of celiac disease has no practical applicability.
TGF-beta signalling pathway factors in HPV-induced cervical lesions.
Iancu IV, Botezatu A, Goia-Ruşanu CD … +5 more , Stănescu A, Huică I, Nistor E, Anton G, Pleşa A
Roum Arch Microbiol Immunol · 2010 · PMID 21434587
Human papillomaviruses (HPV) are considered the etiological agents of cervical cancer, especially high-risk genotypes. TGF-beta (transforming growth factor-beta) is well known for its anti-proliferative effects but the n...
Human papillomaviruses (HPV) are considered the etiological agents of cervical cancer, especially high-risk genotypes. TGF-beta (transforming growth factor-beta) is well known for its anti-proliferative effects but the neoplastic cells often lose their sensitivity to TGF-beta. A characteristic alteration associated with malignant progression is the loss of responsiveness to TGF-beta1-induced cell growth inhibition. The aim of the present study was to establish the possible role of some members of TGF-beta signalling pathway during cervical cancer development and the possible relationship with HPV infection. In order to establish TGF-beta gene expression levels in cervical oncogenesis, TGF-beta1, TGF-beta1 receptors and Smad2 were investigated in precancerous and cervical cancer samples (Quantitative Real-Time PCR). The study revealed that 84.5% of patients were positive for HPV DNA. The most prevalent HPV genotypes were high-risk HPV 16 and 18 in single or co-infections. Expression of TGF-beta1 decreased as tumor cells progressed from cervical intraepithelial neoplasia to cervical carcinoma. Furthermore, we observed that cervical lesions without HPV infection expressed significantly less TGF-beta1. TGF-betaRI and Smad2 gene expression levels were found to be decreased in SCC and AC samples in contrast with CIN1 and CIN2/3 samples. Our results showed that in human cervical cancer the disruption of TGF-beta/Smad signalling pathway might contribute to the malignant progression of cervical dysplasia. These data emphasize the importance of canonical TGF-beta pathway integrity in carcinogenesis.
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