Bacteria-Mediated Delivery of mazF mRNA into Cancer Cells for Induction of Apoptosis

Programmed cell death (PCD), an active process that leads to cell suicide, is a critical mechanism in eukaryotes and prokaryotes. In eukaryotes, apoptosis, a well-characterized form of PCD, is a gene-directed program that can be inhibited by some oncogenic mutations, resulting in the development of cancer. Toxin-antitoxin (TA) is one example of an intracellular death program that is present in almost all bacteria and comprises a stable toxin and a labile antitoxin that blocks the lethal function of the toxin. MazEF, one of the best studied bacterial PCD systems, includes a toxin, namely, MazF that cleaves mRNAs at ACA sequences leading to the inhibition of protein synthesis. Although the exact mechanism remains unknown, it seems the MazF-mediated apoptosis depends on the presence of BAK, a pro-apoptotic protein in mammalian cells. Since cancer is a heterogeneous group of diseases with various sensitivities to different treatments, we examined the impact of MazF proteins on the growth and viability of three cancer cell lines: MCF7, HT29, and AGS. These cells lines were transfected with ACA-less mazF mRNAs, and the cells were evaluated for development of MazF-mediated cell death. Our data illustrated that expression of MazF proteins in the aforementioned cells leads to significant reductions in cell viability. In order to confirm the occurrence of apoptosis, the activation of caspase-3 and -7 and the presence of caspase-3’s substrate were evaluated in transfected cells. Our observations suggested that in the presence of MazF, the levels of activated caspase-3 and -7 were significantly elevated in transfected cells. In addition, the inhibition of protein synthesis by MazF in MCF7 and AGS cell lines were examined via quantifying mRNA translation on a single-cell basis. Our data confirmed that the expression of MazF causes a significant drop in the levels of protein translation in the examined cell lines. Recently, it has been reported that the DNA delivery system does not seem to be a suitable approach to transfer mazF in tumors due to the lack of continuous expression of MazF, degradation, and the incidence of mutations in mazF gene. To address these challenges, we developed an mRNA delivery system to transfer ACA-less mazF mRNA exclusively into cancer cells. To examine the expression and stability of mazF mRNA in cells, we synthesized mazF and GFP mRNA for IRES- or cap- dependent translation. Our results suggest that cap/IRES-dependency, poly-A tails, and modified/unmodified nucleotides played important roles in efficient expression of MazF proteins in transfected cell lines but is dependent on the cell line. MazF is a nonspecific ribonuclease that can induce death in both normal and cancer cells. Thus, the application of MazF is dependent on efficient expression and delivery system to transfer MazF into tumors. We developed a listerial bi-vector expression and delivery system to transfer mazF mRNA into specific HER2-positive SKBR3 human breast cancer cells. Our results showed that following the infection of cells by functionalized bacterial vectors, caspase activities elevated in SKBR3 cells 48 hours post-transfection. In this research, this expression and delivery system for the delivery of mazF mRNA as a potential therapeutic mRNA in cancer cells was reported for the first time

Rupture of Hydatid Liver Cyst into Peritoneal Cavity Following Blunt Abdominal Trauma; a Case Report

Hydatid cyst is a serious parasitic infection in places which people has a close contact with dogs or sheep. They may be found as an incidental findings during routine clinical examination or even followed by radiographic or ultrasonography evaluation. The disease becomes more complicated if rupture of the cysts occurs due to blunt trauma or spontaneously increases of intra-cystic pressure. Here, we reported a case of hydatid liver cyst that ruptured into the peritoneal cavity after blunt abdominal trauma

Antiviral activity of Aloe vera against herpes simplex virus type 2: An in vitro study

In this study we tested the antiviral activity of a crude hot glycerine extract of Aloe vera gel which was grown in Bushehr (Southwest of Iran) against HSV-2 replication in Vero cell line. The extract showed antiviral activity against HSV-2 not only before attachment and entry of virus to the Vero cells but also on post attachment stages of virus replication. The IC50 before attachment and entry of virus to the cells is 428 μg/ml and the CC50 value which is the cytotoxicity of the extract for Vero cells is 3238 μg/ml, while the calculated selectivity index (SI) is 7.56. Also, IC50 of extract on post attachment stages of replication is 536 μg/ml and the SI value for inhibition of the post attachment stages of HSV-2 replication is 6.04. Therefore, compounds of Aloe vera from Bushehr could be a good candidate as a natural source for antiviral drug development against HSV-2

Identification of two Ophiuroidea species in intertidal zone of Chabahar bay along Oman Sea coast

Among the echinoderms, the Ophiuroidea comprise the most diverse class with more than 2,000 species. They occur in nearly any marine habitat, ranging from the poles to the equator and are distributed from the tidal zone down to several thousand meters of depths. Often, their similar morphology makes their species identification difficult. The present study was conducted in order to identify and evaluate the species-spectra of ophiuroids in the Chabahar Bay. Samples were collected from the intertidal zone of Tis port in the west of Chabahar in October 2013, where coarse sediments and pebbles dominate. The samples were examined in the laboratory based on several factors including dorsal and ventral disc and arms. Two representatives of the family Ophiothrichidae were identified: Ophiothrix savgnyi and Macrophiothrix elongata

Assessment of high risk pregnancy in Bam Mahdieh maternity hospital, 2001

Background and aim: Attention to health of mother during pregnancy is essential for the health of both mother and infant. Early detection of high-risk pregnancies is fundamental for prenatal care. This survey was performed to find the prevalence of the high-risk pregnancies and causing factors in the maternity hospital of Bam in 2001. Method: This descriptive and cross sectional study was performed on 850 randomly selected pregnant women. Results: We found that 11.8% of the cases were younger than 18 years, 6.6% were older than 35 years, 8.8% had five or more pregnancies, 5.4% had pre-term, 6.3% had post-term labor, 2% had hemorrhaged pregnancy and 4.2% indicated signs of preeclampsia. The prevalence of various diseases were as follows: 12.3% afflicted with urinary infection, 0.8% had addiction. 0.4% had sexual transmitted disease, 0.4% had anemia, 0.6% had thyroid disease, 0.6% had cardiac diseases, 0.1% had liver disease, 9.1% had diabetes mellitus and 4.3% afflicted with more than one disease. Also 22.9% had low birth weight. Conclusion: Inappropriate age, urinary tract infection, hemorrhage and hypertension were the most important health problems of the pregnant women. For prevention of the high-risk pregnancies, education and prenatal care are necessary

Investigating the effect of brand extensions on brand mental image (Case study: Distribution branches of Minoo Foodstuffs in Mazandaran)

The aim of the present paper is to study the effect of brand extensions on brand mental image distribution branches of Minoo Foodstuffs in Mazandaran during 2013. In this paper, the effects of the related variables were studied such as the effects of mental image of Initial Brand and its fitness with the perceived one based on the brand extensions approach, its fitness with the mental and its effect between initial and final mental image, the effect of mental fitness based on the brand extensions approach and also the brand extensions approach based on the final mental and the perceived fit based on the brand extensions approach. This paper is a functional and measuring study based on its aim and method, respectively. The method used for collecting data is field as well a standardized questionnaire in the papers of Martinez et al. There are 24 questions in this questionnaire, which were arranged in the form of Likert Scale. The statistical method in this paper is structural equations which were analyzed by Lisrel software. In this paper, 7 hypotheses were presented as all of them were confirmed except 5th one

Investigating the effect of brand extensions on brand mental image (Case study: Distribution branches of Minoo Foodstuffs in Mazandaran)

The aim of the present paper is to study the effect of brand extensions on brand mental image distribution branches of Minoo Foodstuffs in Mazandaran during 2013. In this paper, the effects of the related variables were studied such as the effects of mental image of Initial Brand and its fitness with the perceived one based on the brand extensions approach, its fitness with the mental and its effect between initial and final mental image, the effect of mental fitness based on the brand extensions approach and also the brand extensions approach based on the final mental and the perceived fit based on the brand extensions approach. This paper is a functional and measuring study based on its aim and method, respectively. The method used for collecting data is field as well a standardized questionnaire in the papers of Martinez et al. There are 24 questions in this questionnaire, which were arranged in the form of Likert Scale. The statistical method in this paper is structural equations which were analyzed by Lisrel software. In this paper, 7 hypotheses were presented as all of them were confirmed except 5th one