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L-ASPARAGINASE is an anti-neoplasm agent. This agent is classified as an enzyme compound. L-asparaginase is used in medicine as well as in the food industry. Microbial strains that are producing L-Asparaginase can be found in many habitats such as soil. The aim of this study was to find isolates producing L-asparaginase enzyme with high ability to inhibit cancer cells. Bacteria of the Actinobacter family were isolated from the soil samples around the roots of the plants in the actinomycet isolation medium. Then, the bacteria were isolated and extracted by screening in M9 medium. Isolates were identified using16S PCR sequencing method. Three isolates of ASP and B and C showed the highest enzyme production, which showed a molecular weight of 43 kDa and in terms of enzymatic activity112. 572 IU/ml, 99.706 IU/ml, 138.572 IU/ml respectively. These three isolates with sequence analysis was similar to Streptomyces flavoviridis and Streptomyces griseus and Streptomyces hypolyticus respectively. Isolate C showed good effects in inhibiting cell MCF-7, HepG-2. There are various isolates enzyme production power in nature and Actinobacter family has shown more ability to produce various enzymes and metabolites and the isolates isolated in this study are used for industrial enzyme production.

Staphylococcus aureus is the second leading pathogenic cause of nosocomial infections and plays an important role in the development of food poisoning and purulent, systemic and nosocomial infections. This bacterium is responsible for infections such as abscess, food poisoning, third grade burns, traumatic ulcers, surgical incisions, bed sores or atrophic ulcers. Long-term use of antibiotics has led to the emergence of multi-drug resistant strains in infectious diseases. Due to increased resistance of bacteria to chemical drugs and the few side effects of medicinal plants, medicinal plants have nowadays attracted much more attention of researchers. In this regard, studies have been done on the effectiveness of medicinal plants against infectious agents such as S. aureus and methicillin-resistant S. aureus. Therefore, in this review study, native Iranian herbs that are effective on S. aureus were reported. In this review study, search terms including Staphylococcus aureus, herbs, extracts and essential oils were used to retrieve articles indexed in databases such as the Institute for Scientific Information, PubMed, Scopus, Scientific Information Database, Magiran, and Google Scholar. Garlic, pomegranate, Anethum graveolens, coriander, eucalyptus, Lawsonia inermis, Descurainia sophia, Figworts, artemisia, thyme, etc. are some of the most important anti-staphylococcal herbs.

Background: Stem cells are used to treat numerous diseases; however, their lifespan is rather short. Factors such as probiotics affect and improve various cell lineage efficacies. The aim of this study was to investigate the effects of probiotics-conditioned media on dental pulp stem cell potentials in osteogenesis. Methods: The experiment was initiated by culturing Lactobacillus casei and Lactobacillus acidophilus probiotics as well as DPS-7 cells. Bacterial supernatants were separated and concentrated as the conditioned media. The DPS-7 cells were treated with various concentrations of the conditioned media. Furthermore, MTT assay and alkaline phosphatase activity were used. The mRNA expression of three genes (bFGF, EGF-β and BMP-2) involved in osteogenesis was analyzed using a real-time polymerase chain reaction. Results: The response of dental pulp stem cells to probiotics preconditioning promoted cell proliferation, increased alkaline phosphatase activity and upregulated bFGF and BMP-2 gene expression. Increased expression was significant for BMP-2 and moderate for bFGF; however, it was non-significant for EGF-β. The use of the two probiotics was the most effective. Conclusion: In general, synergism of the combined probiotics preconditioning induces differentiation of DPS-7 cells into osteoblasts most effectively.

Background: Helicobacter pylori could colonize the gastric mucosa and cause gastritis, gastric ulcer and gastric cancer. Numerous virulence factors have been identified in this bacterium that play important roles in the promotion of gastric disorders. Although the interaction of long noncoding RNAs (lncRNAs) with transcription, processing, and translation of genes associated with different diseases are described, their interaction with the inflammatory genes and H. pylori infection in the gastric tissue is not well known. In this study, H. pylori infection and its effect on the transcription of common lncRNAs that regulate NF-κB expression were analyzed relative to the non-infected patients. Materials and Methods: Two groups of H. pylori-infected and non-infected patients with chronic gastritis were included in the study after general histopathological and microbiological analysis. Genotyping of the H. pylori strains was done by PCR and relative changes in the expression of NF-κB and regulatory lncRNAs, lincRNA-p21, MALAT1, NKILA, were measured by relative quantitative real time-PCR. Results: Significant increase in the NF-κB gene expression was shown, while the expression level of MALAT1, lincRNA-p21 and NKILA genes decreased in the infected patients compared with the non-infected patients. Our results showed that the hypervirulent strain with oipA“on”/HP-NAP+/iceA1+/iceA2+/vacA s1m1/s1m2+/cagA+ genotype can promote a higher level of NF-κB transcription in the inflamed tissue. Conclusion: H. pylori infection could promote down-regulation of lincRNA-p21, MALAT1 and NKILA in the infected gastric tissue that is correlated with upregulation of NF-κB. More detailed studies are needed to show link between the virulence genes and their impact on the deregulation of lncRNAs in the gastric tissue.

Background: Biofilm formation helps Pseudomonas ‎aeruginosa (P. ‎aeruginosa) survive in various environments. Microgels can be effective in treatment of bacterial infections. The major aim of this study was to investigate effects of poly-N-isopropyl-acrylamide microgels (PNIPAM) on P. aeruginosa. Methods: Totally‎, ‎100 P. aeruginosa strains were isolated from chronic wound infections‎. Quantitative assessments of biofilm formation and antibiotic susceptibility were carried out. Furthermore, algD, lasR, and PA2714 genes were amplified to investigate gene frequencies and expression rates. Results: Significant decreases were seen in lasR expression in EDTA-treated samples‎. Significant decreases were observed in expression of algD and lasR treated with xylitol. Decreased ‎expression of PA2714 was seen in samples treated with xylitol with no significance. ‎ Conclusion: The PNIPAM containing xylitol ‎or EDTA could penetrate biofilms of P. aeruginosa and significantly decrease expression of lasR and algD. This can be a novel strategy in the management of chronic wounds.

Introduction: Bioactive glass (BAG) is increasingly used in dentistry, aiming to provide superior mechanical properties, optimal chemical stability, and favorable antimicrobial activity in the oral environment. This study aimed to measure the flexural strength (FS) and antimicrobial activity of resin-modified glass ionomer (RMGI) cement containing 58S nano-BAG. Materials and Methods: In this in vitro study, 0wt (Weight) %, 10wt%, 20wt%, and 30wt% 58S nano-BAG particles were added to RMGI powder in groups 1 to 4, respectively (n=10). Forty specimens were fabricated in metal molds (2 x 25 x 2 mm), and their FS was measured by using a three-point bending test at a crosshead speed of 0.5 mm/min. The antibacterial activity of the materials against Streptococcus mutants was assessed by the disc diffusion test. In addition to the abovementioned experimental groups, one control group (n=10) containing 100% BAG was also considered. Data were analyzed by one-way ANOVA and Tukey’s test. Results: The mean (± standard deviation) FS was 38.71±8.84, 43.61±17.34, 45.62±15.89, and 54.71±14.25 MPa in groups 1 to 4, respectively. No significant difference was noted in FS among the groups (P=0.06). A significant difference was found in the diameter of the growth inhibition zone among the groups (P Conclusion: The addition of 10wt%, 20wt% and 30wt% nano-BAG to RMGI powder did not significantly change the FS but the addition of 30wt% nano-BAG to RMGI significantly inhibited the bacterial growth.

Background: Primary endodontic infections are caused by necrotic colonization of the pulp tissues by microorganisms. The success of endodontic treatment depends on complete debridement and disinfection of the root canal space. Maximum antibacterial effects, maximum dissolving effects on necrotic tissues, and minimum toxic effects on the periapical tissues are essential characteristics of ideal root-canal irrigants. Recently, researchers have become further interested in medicinal plants as herbal medicines due to their good antimicrobial activities and natural origins. Objectives: This study aimed to compare the antimicrobial properties of chlorhexidine solution alone and mixed with two herbal solutions in root dentine and optimize factors affecting the disinfection of primary tooth root canals. Methods: Teeth infected with pure Enterococcus faecalis suspensions were treated with chlorhexidine solution alone and mixed with chamomile and Satureja khuzestanica. After selecting the most efficient solution, factors possibly affecting the disinfection process were optimized using a central composite design. These factors were the CHX concentration, chamomile concentration, and contact time. Results: The microbial colony count showed that the minimum colony count belonged to 1% chamomile essence and 1% CHX solution, while the maximum colony count belonged to 1% S. khuzestanica and 1% CHX solution, with significant differences. In the optimization process, the best chlorhexidine concentration, chamomile concentration, and contact time were 1.13%, 1.39%, and 11.33 min, respectively. Conclusions: A combination of chlorhexidine and chamomile provided protection against oral microbial pathogens for endodontic infections. However, further comprehensive in vivo studies seem necessary.

Background: Foodborne diseases are a major problem worldwide. The epidemiological investigations in many parts of the world have shown an increase in infections caused by Salmonella serovars. Furthermore, the emergence of drug resistance among them has become a major global concern and awareness of the resistance patterns of Salmonella could be very useful in treatment of diseases. Objective: This study aimed to investigate Salmonella serotypes in foodborne outbreaks by sequencing of ITS region of 16S-23SrRNA gene and to determine their antimicrobial susceptibility pattern. Materials and Methods: A total of 614 diarrheal stool samples were collected from 173 foodborne outbreaks in different provinces of Iran during one year. Identification of Salmonella was carried out by phenotypic and molecular (16s-23srRNA gene detection) methods and antibiotic susceptibility was performed using disc diffusion method. Results: Out of 614 samples, 18 isolates were identified as Salmonella of which 16 (88.9%) isolates were Salmonella Enteritidis and 2 (11.1%) isolates as Salmonella Paratyphi A. All isolates were sensitive to ceftazidime, and high resistance was seen with nalidixic acid with 14 (77.8%) isolates. Conclusion: Increasing antibiotic resistance in many bacterial pathogens such as Salmonella has been a major threat for human health. Therefore, identifying the antibiotic resistance patterns of Salmonella serovars may help in treatment of the associated infections.

Background: Gastritis is among the most common human diseases worldwide. Although the involvement of Helicobacter pylori infection as a class I human carcinogen for gastric cancer progression is accepted, it is not well known how gastritis progression to atrophy and stomach cancer occurs. In this case-control study, the potential link of H. pylori infection with alteration in the transcription of genes involved in DNA Damage Response pathways was investigated among the patients with gastritis. Methods: To measure the difference in relative mRNA expression level of ATM, CHEK2, TP53, DCLRE1C, POLM, XRCC4 genes between H. pylori infected and non-infected patients, gastric biopsies of 30 H. pylori infected patients with moderate chronic gastritis and 30 non-infected patients with mild chronic gastritis were analyzed. Results: Up-regulation of genes linked to non-homologous end joining (NHEJ) pathway (DCLRE1C, POLM, and XRCC) was shown in 40% (8.44 fold ±13.91), 63.33% (15.72 fold ±33.08) and 50% (9.99 fold ±21.55), respectively, and also to DDR pathway (ATM, CHEK2 and TP53) in 33% (2.42 fold ±3.17), 40% (2.86 fold ±3.61) and 50% (5.00 fold ±6.52), respectively. No correlation was detected among alteration in the transcription level of the studied genes and age or gender.Conclusion: Our results provide new data that may support the potential involvement of H. pylori infection in the activation of genes involved in DNA damage response, mainly through non-homologous end joining DNA repair system that might be linked to mutagenesis in the pre-cancerous gastric tissue.

Background and Objectives: Silver nanoparticles (Ag-NPs) are potent antimicrobial agents, which have recently been used in dentistry. The aim of the current study was to optimize antimicrobial activity of Ag-NPs used in preparing irre- versible hydrocolloid impressions against three microorganisms of Escherichia coli, Streptococcus mutans and Candida albicans. Materials and Methods: After assessing antimicrobial activity of the compound using disk diffusion method, three parame- ters of concentration of Ag-NPs (250-1000 ppm), ratio of hydrocolloid impression material powder to water (0.30-0.50) and time of mixing (20.0-60.0 s), affecting antimicrobial activity of irreversible hydrocolloid impression materials against the three microorganisms, were optimized. This combined process was successfully modeled and optimized using Box-Behnken design with response surface methodology (RSM). Decreases in colony number of E. coli, S. mutans and C. albicans were proposed as responses. Results: Qualitative antimicrobial assessments respectively showed average zone of inhibition (ZOI) of 3.7 mm for E. coli, 3.5 mm for S. mutans and 4 mm for C. albicans. For all responses, when the mixing duration and powder-to-water ratio increased, the circumstances (mixing duration of 59.38 s, powder-to-water ratio of 0.4 and Ag-NP concentration of 992 response) increased. Results showed that in optimum ppm, the proportion of decreases in colony numbers was maximum (89.03% for E. coli, 87.08% for S. mutans and 74.54% for C. albicans). Regression analysis illustrated a good fit of the ex- perimental data to the predicted model as high correlation coefficients validated that the predicted model was well fitted with data. Values of R2Adj with R2Pred were associated to the accuracy of this model in all responses. Conclusion: Disinfection efficiency dramatically increased with increasing of Ag-NP concentration, powder-to-water ratio and mixing time.

Background: Tooth decay is an infectious disease that is caused by various microorganisms. Chemicaldrugs used to control and treat tooth decay mainly have side effects. On the other hand, many plants haveantimicrobial effect against microbes that cause tooth decay. Therefore, the aim of this study was to introduceIranian medicinal plants with antimicrobial properties in tooth decay.Method: To reach this purpose, we searched keywords containing Iranian medicinal plants, medicinalplants, Tooth decay, essential oils, and extracts in Pub Med, Web of Science Direct, Scopus, ScientificInformation Databases, and Google scholar.Results: The results of our study led to the achievement of 17 articles that intensive on the study of Iranianmedicinal plants affecting the bacteria that cause tooth decay.Conclusion: This article introduces a number of plants that have been introduced in different regions of Iranfor the treatment of tooth decay, which based on the knowledge of traditional and ethnobotanical medicineof Iran, we believe that they can be used as a drug against tooth decay.

Background: Klebsiella pneumoniae is an opportunistic pathogen causing nosocomial infection in human. This study aimed to investigate the relationship between the presence of genes involved in biofilm formation in K. pneumoniae isolated from patients and the presence of antibiotic resistance genes. Methods: Biochemical tests were used for the identification of K. pneumonia isolated from urine samples referred to hospitals in Tehran, Iran, from Sep 2018 to Jan 2020. The antibiotic resistance pattern was performed and biofilm formation was assessed phenotypically. Finally, β-lactamase genes and adhesion genes were detected by the PCR method. Results: We collected 457 K. pneumoniae isolates from hospitals in Tehran, Iran. 110 isolates were resistant to imipenem. Fifty isolates were positive for metallo-β-lactamases that thirty-nine isolates (35.45%) has blaKPC gene, 18 isolates (16.36%) had blaVIM-1 gene and 9 isolates (8.18%) had blaIMP-1 gene detected by PCR. Sixty isolates (54.54%) had strong biofilm, 35 isolates (31.81%) had moderate biofilm and 15 isolates (13.63%) had weak biofilm. The presence of adhesion genes in K. pneumoniae isolates significantly correlated with resistance genes (P

Background Helicobacter pylori infection and heterogeneity in its pathogenesis could describe diversity in the expression of inflammatory genes in the gastric tissue. We aimed to investigate transcriptional alteration of genes linked to gastritis concerning the H. pylori infection status and its virulence factors. Methods and Results Biopsy samples of 12 infected and 12 non-infected patients with H. pylori that showed moderate chronic gastritis were selected for transcriptional analysis. Genotyping of H. pylori strains was done using PCR and relative expression of inflammatory genes was compared between the infected and non-infected patients using relative quantitative real-time PCR. Positive correlations between transcriptional changes of IL8 with TNF-α and Noxo1 in the infected and TNF-α with Noxo1, MMP7, and Atp4A in the non-infected patients were detected. Six distinct genotypes of H. pylori were detected that showed no correlation with gender, ethnicity, age, endoscopic findings, and transcriptional levels of host genes. Irrespective of the characterized genotypes, our results showed overexpression of TNF-α, MMP7, Noxo1, and ATP4A in the infected and IL-8, Noxo1, and ATP4A in the non-infected patients. Conclusions A complexity in transcription of genes respective to the characterized H. pylori genotypes in the infected patients was detected in our study. The observed difference in co-regulation of genes linked to gastritis in the infected and non-infected patients proposed involvement of different regulatory pathways in the inflammation of the gastric tissue in the studied groups.

BACKGROUND: Pseudomonas aeruginosa is an opportunistic human pathogen that causes severe acute and chronic nosocomial infections, especially in immunocompromised burn patients. and can lead to severe mortality and morbidity. The emergence of antibiotic resistant P. aeruginosa infections has created significant challenges in treating these patients. A potential alternative treatment for antibiotic resistant pathogens includes the use of carbon nanotubes (CNTs), which have received considerable attention due to their potent antibacterial activity. The aim of this study was to construct a novel CNT containing an anti-bacterial chemical component to effectively combat drug resistant P. aeruginosa infections. METHODS: In this study, a novel chemical component was synthesized and coated the CNT. The antimicrobial effects were then evaluated on MDR, XDR, and PDR strains of P. aeruginosa isolated from burn patients. Antibiotic susceptibility was evaluated using the disk diffusion test and minimum inhibitory concentration (MIC) testing. In order to determine the potential cytotoxicity, an MTT assay was performed on Human Dermal Fibroblasts. The effect of treatment on the expression of wound healing genes was analyzed via qRT-PCR. RESULTS: Experimental data indicates that our CNT coated chemical compound had antibacterial properties, negligible cytotoxicity, and could accelerate the wound healing process. CONCLUSION: Given the antibacterial properties of our CNT chemical compound, it has the potential to treat and reduce the occurrence of multi-drug resistant P. aeruginosa burn wound infections and aid in wound healing by turning on genes (VEGFA, EGF and PDEGF) involved in the wound healing process.

Ciprofloxacin is an alternative to vancomycin for treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections. The objective of this study was to optimization of niosomes encapsulated ciprofloxacin and evaluate their antibacterial and anti-biofilm efficacies against ciprofloxacin-resistant methicillin-resistant S. aureus (CR-MRSA) strains. Formulation of niosomes encapsulated ciprofloxacin were optimized by changing the proportions of Tween 60, Span 60, and cholesterol. The optimized ciprofloxacin encapsulated niosomal formulations based on Span 60 and Tween 60 were prepared and characterized using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and dynamic light scattering (DLS). The SEM and TEM results showed that the formulation of niosomes encapsulated ciprofloxacin were spherical with a size between 50 and 150 nm. The prepared niosomal formulations showed high storage stability up to 30 days with the slight change in size and drug entrapment during the storage, making them good candidates for drug delivery systems. Optimum niosome encapsulated ciprofloxacin enhanced antibacterial activity against CR-MRSA strains via reduction in minimum inhibitory concentration (MIC) value and inhibited significantly biofilm formation. Niosome encapsulated ciprofloxacin down-regulated the expression of icaB biofilm formation gene. Our results showed that encapsulating ciprofloxacin in niosomes is a promising approach to enhanced antibacterial activity, biofilm inhibition and reduced resistance to antibiotic in CR-MRSA strains.

Background Dental caries is considered the most common infectious disease in humans worldwide. Cariogenesis is the outcome of a complex interaction between the host's oral flora and diet. The consumption of snacks such as cake, which have the potential to promote dental caries, has increased. Objectives The aim of this study was to investigate the effect of including probiotic bacteria (Bacillus coagulans - B. coagulans) in consumed snack cake on the Streptococcus mutans (S. mutans) count and salivary pH. Material and methods We conducted a randomized, double-blind, cross-sectional cohort study on 40 healthy volunteers. The subjects were divided into 2 groups. In the 1st group, the subjects consumed probiotic cake as breakfast for 1 week and then, following a 4-week wash-out period, consumed regular cake as breakfast for 1 week. In the other group, the administration of probiotic and regular cake was reversed. For both groups, samples of at least 5 mL of non-stimulated saliva were collected using the spitting technique before and after the 1st and the 6th week. A colony counter was used to determine the number of S. mutans colonies. Salivary pH was measured before eating (8-9 a.m.). Results We detected no statistically significant difference in the S. mutans count before and after the consumption of probiotic cake, but noted a statistically significant difference in the count before and after the consumption of regular cake. We did not detect a significant difference in salivary pH with respect to the consumption of probiotic and regular cake, although the consumption of both foods caused a drop in salivary pH. Conclusions The addition of probiotic bacteria to sweet snack cake caused a minimal increase in the salivary count of S. mutans, a bacterial species with a definite role in cariogenesis, but did not impact salivary pH. Since probiotic cake has a slight impact on the S. mutans count, it is preferred over regular cake as a snack food.

Oral squamous cell carcinoma (OSCC) are one of the major causes of cancer morbidity and mortality worldwide. Dental microbiome has been considered as inducing agents in oral carcinogenesis. Therefore, the objective of this study was to investigate the interaction of the gene expression of the dental microbiome and OSCC patients. A cross-sectional study was designed by recruiting confirmed OSCC patients attending the University hospital during October 2018 and July 2019. The dental bacteria were isolated and confirmed by PCR technique. The expression of host and bacterial virulence genes was determined using qPCR. This study shows that 54% of T. forsythia found to be the most predominant organisms in 30 positive cases, followed by 34% of Campylobacter rectus and 29% of Prevotella intermedia. The expression of mRNA levels of bspA, csxA, fadA and interpain A in the OSCC- bacteria positive cases was significantly higher than the control group (P 0.001). It was further found that interpainA, csxA, fadA, and bspA genes have the potential effects on the cellular gene expression in OSCC patients. A significant correlation was seen between expression patterns of CXCL10, DIAPH1, NCLN and MMP9 genes with interpain A, fadA, and bspA involved in OSCC cases The results indicate that the species specific bacteria may play a role in triggering chronic inflammation in OSCC patients. Therefore, alteration in the gene expression through the dental microbiome could be used as an alternative target in the clinical practice to detect OSCC.

A total of 100 Staphylococcus aureus strains were isolated from 1,047 food samples. In addition to biotyping all isolates, the occurrence of the tst gene and protein were evaluated by PCR and the RPLA test, respectively. Moreover, polymorphism of the X-region of the protein A gene was analyzed by PCR-RFLP. TSST1 was detected in 12 strains and production of TSST1 in all strains was confirmed by RPLA assay. It was noteworthy that 66.7% of TSST1-producing S. aureus strains belonged to the human biotype. The result of genotyping amplification of the spa gene displayed size polymorphisms and revealed seven different clusters ranging from 1,200 bp to 1,600 bp. Digestion of amplicons with HindIII and HaeII resulted in six distinct patterns for each of them. An amplicon of 1,450 bp in size was the predominant type of Spa-PCR product. The accuracy of the Spa typing system by PCR-RFLP was 100%, thus confirming this method as a reliable tool in routine epidemiological investigation for S. aureus.

Resistance to b-lactam antibiotics by gramnegative bacteria, especially Escherichia coli (E. coli), is a major public health issue worldwide. The predominant resistance mechanism in gram negative bacteria particularly E. coli is via the production of extended spectrum beta lactamase (ESBLs) enzymes. In recent years, the prevalence of b-lactamase producing organisms is increased and identification of these isolates by using disk diffusion method and no-one else is not satisfactory. So, this investigation focused on evaluating the prevalence of ESBL enzymes by disk diffusion method and confirmatory test (Combined Disk). Five hundred clinical samples were collected and 200 E. coli isolates were detected by standard biochemical tests. To performing initial screening of ESBLs was used from Disk diffusion method on E. coli isolates. A confirmation test (Combined Disk method) was performed on isolates of resistant to cephalosporin's indicators. Up to 70% isolates exhibited the Multi Drug Resistance phenotype. In Disk diffusion method, 128(64%) E. coli isolates which resistant to ceftazidime and cefotaxime while in Combined Disk, among 128 screened isolates, 115 (89.8%) isolates were detected as ESBLs producers. This survey indicate beta lactamase enzymes are playing a significant role in antibiotic resistance and correct detection of them in phenotypic test by using disk diffusion and combined Disk is essential for accurate recognition of ESBLs.

Background: Antimicrobial peptides play a role as effectors substances in the immunity of vertebrate and inverte­brate hosts. In the current study, antimicrobial peptide was isolated from the haemolymph of the American cock­roach, Periplaneta americana. Methods: Micrococcus luteus as Gram-positive bacteria and Escherichia coli as Gram-negative bacteria were candi­date for injection. Induction was done by injecting both bacteria into the abdominal cavity of two groups of cock­roaches separately. The haemolymphs were collected 24 hours after post injection and initially tested against both bacteria. Subsequently, the immune induced haemolymph was purified by high performance liquid chromatography (HPLC) to separate the proteins responsible for the antibacterial activity. Results: The non-induced haemolymph did not show any activity against both bacteria whereas induced haemo­lymph exhibited high activity against M. luteus but did less against E. coli. Two fractions showed antibacterial activ­ity against M. luteus. Finally the molecular weight of the isolated antibacterial proteins were determined as 72 kDa and 62 kDa using SDS-PAGE. Conclusion: Induced haemolymph of American cockroaches has the ability to produce peptides to combat against Gram-positive bacteria when an immune challenge is mounted. Further work has to be done to sequence of the pro­tein, which it would be advantageous.

Background: Uropathogenic Escherichia coli (UPEC) are major bacterial agent of Urinary Tract Infection (UTI). This infection is more prevalent among women because approximately half of all women will experience a UTI in their life-time and near a quarter of them will have a recurrent infection within 6–12 months. IutA protein has a major role during UPEC pathogenesis and consequently infection. Therefore, the aim of current study was assessment of IutA protein roles as a potential candidate antigen based for vaccine designing.Methods: This survey was conducted during 2014-2015 at the University of Tehran, Iran. Chromosomal DNA extracted from E. coli 35218 and iutA gene amplified by PCR. The amplicon cloned to pVax.1 eukaryotic expression vector and recombinant vector confirmed by sequencing. The iutA gene expression in genetic cassette of pVax/iutA was evaluated in COS7 cell line by RT-PCR. Then, injected to mouse model, which divided to three groups: injected with pVax vector, PBS and pVax/iutA cassette respectively in two stages (d 1 and 14). One week after the second injection, bleeding from immunized mouse was performed and IFN-gamma was measured.Results: The mice immunized with pVax/iutA showed increased interferon-γ responses significantly higher than two non-immunized groups (PConclusion: Cellular immune response has a main protective role against UTI. Raising this kind of immune response is important to preventing of recurrent infection. Moreover, the current DNA cassette will be valuable for more trying to prepare a new vaccine against UTI. Keywords: Genetic vaccination, Uropathogenic escherichia coli, IutA