International Journal of Molecular and Clinical Microbiology
Volume:11 Issue: 2, Summer and Autumn 2021

Mussels secrete protein-based polymers mainly mussel foot proteins (Mfps), enabling them to anchor to various surfaces in a saline, intertidal zone. Using Mfp proteins as novel water-resistant adhesive materials have been suggested for biomedical purposes due to their unique features including large abundance of catechol aligned with amphiphilic and ionic properties. Despite the more promising characteristics of foot proteins, they have not been widely exploited for biomedical or even industrial applications, since farming the mussels are not commercially viable due to ecological issues and their highly territorial nature. Present study aimed to examine engineering of recombinant synthesis of Mfp3 protein fused to another marine based curli protein, gas vesicle protein A(GvpA) in nonpathogenic yeast, Pichia pastoris, expression system. To this end, sequences of the Mfp3 and GvpA were extracted from the NCBI database and were inserted into the pPICZ A vector in order to obtain an efficient expression level of chimeric protein. Obtained vectors were transformed into E. coli TOP10F' for multiplication and then linearized plasmid transformed into Pichia pastoris for protein expression. The best expression level obtained after 96 hours incubation with methanol induction. In conclusion, active recombinant adhesive Mfp3-GvpA fused protein were successfully expressed in P. pastoris suggesting potential expression system for future bio-adhesive or any fused protein production.

Recently, fungal drug resistance has significantly increased especially in opportunistic fungus like Candida albicans. Accordingly, it is necessary to use more effective drugs with less toxicity and high influences. Among a large of investigations, ionic liquids showed biological influence and antimicrobial activities. The aim of this study was in vitro investigation of antifungal activity of pyridine –based ionic liquid on a standard strain of Candida albicans and evaluate its toxicity on host cells. A standard strain of Candida albicans was re- cultured on Sabouraud Dextrose Agar (SDA) containing chloramphenicol, incubated at 37℃ for 24 h. Antifungal effect of a novel ionic liquid ([Met-Hcl] [Pys] ), was evaluated using inhibitory zone diameter. The minimum inhibitory concentration (MIC) and minimum fungal concentration (MFC) were performed using micro dilution method. In continue MTT test were done to evaluate the toxicity of the liquid on host cells. The ionic liquid showed a good effect to prevent of Candida albicans growth. Inhibitory zone diameter was between 34±1 mm. The MIC evaluation was 708.4 ppm. Also the results of MTT test showed the viability of host cells at the 16 dilution. In conclusion, the results of this study manifested that the novel ionic liquid has a good antifungal activity against Candida albicans standard strain with low toxicity to human cells and probably can use as a good novel drug in treatment of candidiasis. Although the need for more studies is crystal clear.

Pseudomonas aeruginosa is an opportunistic nosocomial infection implicated in bacteremia in patients with compromised host defenses. Resistance to ciprofloxacin and imipenem, which are considered as suitable therapeutic options, is increasing in P. aeruginosa. Curcumin is a diferuloylmethane with antimicrobial properties. The present study was conducted to find the molecular effects of curcumin on clinical isolates with mutated genes involved in ciprofloxacin resistance. Fifty-two clinical isolates of P. aeruginosa were obtained from several hospitals and laboratories in Guilan province, northern Iran. Susceptibility to five antibiotics was evaluated by disc diffusion and broth dilution (MIC) methods. Furthermore, PCR-sequencing was carried out to evaluate mutations in topoisomerase subunits, five negative regulators of efflux pumps and oprD gene in these isolates. The effects of curcumin on the expression of mexB and mexY were evaluated using Q-RT-PCR.Of 52 P. aeruginosa isolated strains, 32-44% resistance to amikacin, ciprofloxacin, imipenem and gentamicin was observed. All isolates had mutation in gyrA. Some isolates had mutation in other topoisomerase subunits, some negative regulator genes and oprD gene. Curcumin (400µg/ml) along with ciprofloxacin (subMIC) increased ciprofloxacin susceptibility in four isolates. In these isolates, the expression of MexB and MexY efflux pump genes were downregulated. It seems that among P. aeruginosa isolates with various mutations in important genes in antibiotic resistant pathways, curcumin can intelligently sensitize isolates to these drugs.

Candida albicans (C. albicans) is responsible for most invasive fungal diseases and colonized on the skin and mucosal membranes. The fungus is a part of human natural microflora. Recently, increasing the resistant strains of C. albicans led the researchers to search for new drugs in treatment of candidiasis. The aim of this paper was to investigate the antifungal activity of ([prolinium chloride] [1-methylimidazolium 3-sulfonate]) on a candida albicans standard strain. To reach this purpose, minimum inhibitory concentration (MIC) were performed via micro dilution methods. Then to understand the toxicity of the IL, MTT test was done. The obtained data showed that the IL can inhibit C. albicans growth at the 1690 mg/ml concentration (P<0.0001). Also the IL showed low toxicity to mammalian cells at the same concentration (P<0.005). Our results showed that the IL [prolinium chloride] [1-methylimidazolium 3-sulfonate] has a good antifungal activity with low cytotoxicity that may be a good candidate for new drugs designing.

Cancer is one of the leading causes of death worldwide, with approximately 10 million people dying by 2020. Antibiotics are becoming increasingly ineffective as drug resistance spreads around the world, making infections and death more difficult to treat. Natural resources play an important role in the development of anticancer and antimicrobial agents. The aim of this study was to evaluate the anticancer and antibacterial effect of Avena ludoviciana L. leaves. Chemical compounds were screened and identified using GC mass spectrometry. The anti-cancer effect of hydroalcoholic extract and fractions (hexane, chloroform, and ethyl acetate) of Avena ludoviciana L. leaves were evaluated by the MTT method on Skov3 and MRC5 cell lines. Antibacterial activity of hydroalcoholic extract of Avena ludoviciana L. leaves on four bacterial strains was investigated by agar well diffusion method and the minimum inhibitory concentration was determined by dilution methods. The results showed that different concentrations of hydroalcoholic extracts and fractions significantly reduced the growth of the Skov3 cell line compared to the control group after 48 hours, dose-dependently (P <0.05). Hydroalcoholic extracts except E. coli were tested on all gram-positive and gram-negative bacteria (P <0.05). The largest growth-inhibitory diameter was observed in S. aureus. that was the most sensitive bacteria (lowest MIC) and B. cereus was the most resistant bacteria (lowest MIC) to the extract. Our results show that medicinal plants can be promising sources of natural products with potential anticancer and antimicrobial activity. Further research is suggested for clinical trials, identification, and extraction of effective compounds.

Take-all and common root rot diseases are the important diseases of wheat that are caused by two fungi, Gaeumannomyces graminis var. tritici and Bipolaris sorokiniana, respectively. In this study, we aimed to isolate rhizobacteria Bacillus and Pseudomonas from wheat rhizosphere and evaluate their antagonistic effect against G. graminis var. tritici and B. sorokiniana. Thirty six soil samples from wheat rhizosphere were cultured and isolates were identified by bacteriological and biochemical tests. Using the dual culture method antagonistic effects of all the Bacillus and Pseudomonas isolates were tested against the target fungal pathogens. The isolates were also evaluated for volatile metabolites and sidrophore production. The polymerase chain reaction assay was performed for accurate identification of the isolates. Fifty seven Bacillus and Pseudomonas strains were isolated. Of the isolates, 19 strains had antagonistic effect against the tested pathogens. Bacillus isolates had a greater antagonistic effect against the tested fungi than Pseudomonas isolates. In addition, Bacillus isolates showed a greater antagonistic effect against B. sorokiniana than G. graminis var. tritici. In this study, only 7 isolates were able to produce volatile metabolites. Sidrophore production was detected in 3 strains of Pseudomonas isolates. Based on the 16S rRNA analysis, the 3 strains of Bacillus and Pseudomonas were identified as Bacillus megatrium, Bacillus subtilis and Pseudomonas aeruginosa. According to majority of the isolates belonged to the Bacillus strains and some of them had a good antagonistic activity against G. graminis var. tritici and B. sorokiniana, they are promising for biocontrol of these important pathogens.

Human blood parasites are one of the most critical infections in human that transmit by vectors. Reservoirs of the parasites are crucially important in the epidemiology and control. In the current study isolated parasites from a Rhombomys Optimus (R. opimus), rodent confirms that Crithidia is a zoonotic parasitic disease. This study aimed to find the high-risk areas of this infection by considering the distribution of reservoirs and human infection. In this study, 148 rodents from an endemic focus of Gonbad-e-Qabus city in Golestan province were trapped and then killed ethically and direct smear and culture in Novy- MacNal-Nicolle medium (NNN) were taken and finally, results were confirmed by Polymerase Chain Reaction (PCR) and Sequencing method. Out of 148 rodent, 97 (65.54%) rodent were male and 51 (34.45%) were female (P <0.05). and in smear and culture were found 8 (5.40%) T. lewisi, 6 (4.05%) L. major, and 2 (1.35%) Crithidia spp. Based on the time; 40 (27.02%), 50 (33.78%), 38 (25.67%), and 20 (13.51%) rodents were trapped in spring, summer, fall, and winter, respectively. Due to northeastern Iran (Gonbad-e-Qabus) being the endemic focus of cutaneous leishmaniasis (CL), it should be noted that the reservoirs of this disease may also be contaminated with Leishmania spp, and Crithidia. Results showed that R. opimus are the important reservoirs of CL in northeastern of Iran. Important foci of the diseases in almost all areas of Iran are dispersed. Therefore, reliable methods to control mice are essentially needed.

Natural plant products are the best candidate for antimicrobial and antioxidant properties and are the suitable alternative for chemical drugs. This study aimed to examine the antimicrobial effect of ethanol extract of S. officinalis on S. aureus, E. coli, K. pneumoniae and P. aeruginosa and its comparison with antibiotic discs of ciprofloxacin, ceftriaxone and gentamicin. In this experimental study the ethanolic extract of S. officinalis was extracted by maceration method and the concentrations of 1.9, 3.9, 7.8, 15.6, 31.2, 62.5, 125, 250, 500 and 1000 µg/ml were obtained. Standard microbial strains of S. aureus, E. coli, K. pneumoniae and P. aeruginosa were purchased from pasture Institute and the amount of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extracts was determined using micro dilution method. Antioxidant and cytotoxicity were evaluated using Diphenyl-1-Picrylhydrazyl (DPPH) Radical Scavenging Capacity and MTT assays. The ethanol extract of S. officinalis had different effects on S. aureus, E. coli and P. aeruginosa strains and inhibition zone diameter of 32.66, 10.83 & 10.6 mm were observed respectively. S. officinalis had an inhibitory effect on all of the studied bacteria except K. pneumoniae and this effect was higher in S. aureus bacteria. Also, the inhibition zone diameter of S. officinalis extracts was exceptionally higher in S. aureus compared to ceftriaxone and gentamicin. Moreover the ethanol extract of S. officinalis showed acceptable antioxidant and no cytotoxicity effects. Our results indicated that S. officinalis extracts had the greatest antibacterial effect on the gram-positive bacteria. Although the inhibition zone diameter of S. officinalis extracts was exceptionally higher in S. aureus compared to ceftriaxone and gentamicin

The occurrence of gastric cancer is associated with numerous aspects, including the host's lifestyle and genetic history. Understanding gastric cancer molecular mechanisms can improve our insight into the early diagnosis, prognosis, and treatment. In this study, the RNA level of SNHG8, AF147447, and n34560 genes in gastric tumor tissues was investigated and their association with Helicobacter pylori and Epstein-Barr virus infections was evaluated. Formalin-fixed paraffinembedded (FFPE) tissues (100 samples), including 50 samples of gastric cancer tissues and 50 samples of healthy tissues were taken. The expression level of SNHG8, AF147447, and n34560 genes in gastric cancer and control tissues were examined using the qRT-PCR technique. A significant association was observed between the expression level of the SNHG8 gene in gastric tumor tissues compared to the healthy tissues (P=0.0003). Relative expression of AF147447 and n34560 genes did not show any significant difference among gastric tumor tissues compared to the normal tissues (P=0.2984, P=0.9158). In addition, pathological comparison of clinical data with the expression of SNHG8, AF147447, and n34560 genes did not show any significant association in tumor and healthy tissues, but the expression level of AF147447 gene in Helicobacter pylori infection (P=0.0458) and expression level of n34560 gene in Epstein-Barr virus (EBV) infection (P=0.0362) showed significant association. In conclusion, we found a significant association between SNHG8 gene expression levels and the possible cancer incidence. Also, a significant association was observed between the expression of n34560 and AF147447 genes relating to H.pylori and EBV infections in gastric cancer.

Mixed vaginitis is the simultaneous presence of two or more types of pathogens. Zinc oxide (ZnO NPs) is commonly used in pharmaceutical products. Our study investigates the role of ZnO NPs and vitamin C (VC) in resolving mixed vaginitis. The NMRI mice were inoculated with a mixture of Candida albicans and Escherichia coli. Mice were classified into 8 groups: (1) control, (2) intact mice that received ZnO NPs, (3) intact mice that received daily injection of VC, (4) intact mice that received co- administration of ZnO NPs and VC, (5) infected, (6) infected treated with ZnO NPs, (7) infected that received daily injection of VC, and (8) infected mice treated with co- administration of ZnO NPs and VC. The antimicrobial activity was evaluated using broth dilution methods. Blood samples were obtained for hematological analysis. Vaginal tissue samples were separated and histopathological analysis was performed. Co-administration of ZnO NPs and VC improved the hematological profiles and vaginal architecture. Inhibitory concentration (IC- 50 and IC- 90) of ZnO NPs for the mixture of C. albicans and E. coli were 235.86 and 685.81 ppm, respectively. Co-administration of ZnO NPs and VC in mixed vaginitis management, Mixed vaginitis disrupted the structure of the vaginal epithelium. Consumption of ZnO NPs also caused adverse changes in the structure of the vagina, but co-administration of nanoparticles and VC completely improved the negative effects of infection and ZnO NPs in vaginal tissue, that these agents can be used in the management of mixed vaginitis.