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Cancer is the uncontrolled division of cells and can be caused by genetic or environmental factors. Pancreatic cancer is one of the deadliest among all cancers. The role of bacteria as an anticancer agent dates back to almost 100 years ago. The microbiome has recently become a focus of research in carcinogenesis and even pancreatic cancer. Shigella flexneri is a gram-negative bacterium, which causes shigellosis with symptoms such as diarrhea, fever, and stomach cramps in human. Shigella flexneri may play a very important role in the internal pathways of apoptosis and may induce apoptosis in some of the cancerous cells. In this experiment bacteria were cultured on Salmonella-Shigella agar, then inoculated into BHI Broth medium. After sonication, the protein concentration of the bacterium was measured by using the ZellBio Sensitive Protein Bradford Assay kit. MTT assay was performed to obtain IC50 for the said bacterial protein. Later by cDNA kit synthesized the cDNA based on the RNA template. In the end, the results were analyzed using real-time PCR and the expression of Bax and Bcl-2 genes was measured before and after treatment. The results showed that Shigella flexneri has the potential anti-proliferative effect in pancreatic cancer. The inhibitory concentration, pro-apoptotic amount to upregulate Bax, and meanwhile also to downregulate the bcl-2 found to be 10 μl. In general, due to defects in the apoptotic pathway in cancer cells and the existence of drug-resistant cells, the detection of new apoptotic inducers such as Shigella flexneri cell extract can be used for further studies on cancer therapy.

IntroductionThe top 3 causes of death worldwide include heart disease, injury, and cancer; and cancer records the 2nd place as the leading cause of death in the United States of America after cardiovascular diseases and injuries. Cancer can begin and progress in a very highly twisted and complex pattern and follow the multifactorial route. There is only very few research on medicinal properties Oliveria decumbens rare and valuable plant specially on cancer. So, in this study we tried to cover all needs for future in vivo research.MethodsMTT assay has been performed to estimate the cytotoxicity of the ethanolic extract of the plant. Its free radical capacity evaluation was done by DPPH assay. Furthermore, real-time PCR, the wound-healing assay along with a DNA damage test to study DNA fragmentation characteristics. The plant’s transcriptomic study was performed by NGS de Novo assembly.ResultOliveria decumbens ethanolic extract showed an Ic50 of 14.39 μg/ml. The real-time PCR showed that Oliveria decumbens ethanolic extract significantly induced apoptosis by upregulating the bax gene and slight downregulation of bcl2 an anti-apoptosis gene. The NGS de Novo transcriptome analysis discovered 38 genes responsible for secondary metabolite synthesis so far. The remaining genes and reconstruction of the co-expression network of the transcriptome are underway.ConclusionThe outcome of the Scratch-test and DNA fragmentation confirmed the anti-metastatic and DNA damage properties respectively. Based on these findings; Oliveria decumbens ethanolic extract shall be considered as potential anticancer and chemotherapeutic agents which may elucidate in upcoming studies.

The most common chronic bacterial infection is Helicobacter pylori. The connection between chronic H. pylori infection and gastric cancer is recognized. The early detection of gastric cancer improves survival. miRNAs regulate gene expression in eukaryotes by inhibiting mRNA translocation or degradation. The objective of this study was to compare the expression of miRNA-17-3p and miRNA-17-5p genes in gastric cancer patients with Helicobacter pylori infection. Herein, 30 isolates were identified as H. pylori based on urease test, and 30 and 12 cases were isolated from gastric cancer patients and non-Helicobacter pylori cases as control, respectively. A peripheral blood sample was collected from patients. Analysis of total mRNA extracts from peripheral blood samples, for gene expression changes (miRNA-17-3p and miRNA-17-5p) by quantitative real-time polymerase chain reaction (qRT-PCR), was done. As said by the results, p values showed that expression levels of miRNA-17-3p and miRNA-17-5p were significantly higher in H. pylori-positive GC patients and H. pylori-positive non-GC patients with comparing by healthy controls. So, there was no significant difference between expression levels of miRNA-17-3p and miRNA-17-5p in H. pylori-positive GC patients and H. pylori-positive non-GC patients. Considering our results, the high expression of miRNA-17-3p and miRNA-17-5p has a direct relationship with increased cell proliferation, inhibition of tumor cell apoptosis and tumor angiogenesis, in addition to miRNAs play an important role as biomarkers in helping for detection of the patient by H. pylori infection to become cancerous. Therefore, it can be used to make specific diagnostic kits and to treat patients.

Background: Pancreatic cancer is one of the most common types of cancer that affects the gastrointestinal tract and the induction of apoptosis in cancer cells is generally one of the suitable methods for treating cancer. Salmonella typhi induces apoptosis by activating the caspase pathway. Methods: This experimental study was conducted on a standard strain and the bacteria were cultured on an SS-Agar medium. Then, sonication was used to isolate bacterial proteins and ZellBio was utilized to determine the concentration of such proteins. In addition, 1 million cells per well were cultivated in the cell culture plate in order to treat the cell line with bacteria and to examine the expression of the Bax and Bcl-2 genes. Then, RNA was extracted and cDNA was synthesized after 24 hours of incubation. The Bax and Bcl-2 genes were expressed by Real-time polymerase chain reaction (RT-PCR). Results: The findings confirmed the role of Salmonella typhi proteins in inducing apoptosis in a pancreatic cancer cell. In other words, the concentration of 10 mg Salmonella typhi proteins increased the expression of the Bax gene while reducing the expression of the Bcl-2 gene. Conclusions: Overall, bacterial treatments have unique mechanisms for treating cancer. For example, bacteria can specifically target the tumors, actively penetrate into the tissue, and potentially form a controlled form of toxicity. Accordingly, further studies are recommended to investigate the role of bacteria, especially Salmonella in the treatment of cancer cells.

Methicillin-resistant Staphylococcus aureus is a problem in both healthcare institutions and community settings. This is due to its multi-drug-resistant challenges. Recent advancement in nanotechnology has expanded our ability to design and construct nanomaterials to treat bacterial infections. Carbon nanotubes are one of these nanomaterials. Herein, twenty isolates from hospitals were identified as S. aureus based on morphology, Gram stain, catalase test, coagulase test, and mannitol salt agar fermentation. Isolates were subjected to bio-typing, the methicillin resistance was characterized using the disk diffusion method and PCR of the mecA gene. Then, the single-walled carbon nanotubes (SWCNTs) were functionalized and conjugated to methicillin. The prepared nano-antibiotics were characterized using FTIR and SEM analysis. The characterization of the nano-antibiotics confirmed the successful functionalization of the SWCNTs and the addition of carboxyl groups. Interestingly, the results of the antibacterial activity indicated that the functionalized methicillin-loaded SWCNTs have a significant increase in the antibacterial activity against the S. aureus in comparison to the methicillin-free drug.

In recent years, CD surface modification methods are employed for immunoassay techniques that is called BioCD technology. In this research, first polycarbonate surface was activated with UV ozone and a hydrophilic surface was obtained. Contact angle measurements and atomic force microscopy technique confirmed the hydrophilic property of surface. After that, tetanus toxoid was immobilized on modified CD surface then specific monoclonal antibody, gold nanoparticles conjugated antibody, silver salt, and hydroquinone were added on modified CD surface. So a sandwiches complex as tetanus toxoid, tetanus toxoid monoclonal antibody, and gold nanoparticles conjugated antibody was obtained on CD surface. ATR result showed the immobilization of tetanus toxoid on modified CD surface. Localized surface plasmon resonance (LSPR) and DLS results confirmed the complex formation. Silver salt and hydroquinone were added for signal amplification. Detection limit of anti-tetanus toxoid IgG monoclonal antibody was obtained 0.005 IU/ml by LSPR and DLS techniques. The presented method increases the assay’s sensitivity. BioCD-based immunoassay for detection of anti-tetanus toxoid IgG monoclonal antibody could be applicable in development and fabrication of biomedical devices.

Background: Parathyroid Hormone (PTH) is secreted by parathyroid glands and controls the level of calcium in bones and kidney. PTH is a small polypeptide with 84 amino acids, but the first 34 amino acids of which are enough for hormone biological activity and can be used in the treatment of Osteoporosis. The expression efficiency of recombinant human parathyroid hormone rhPTH (1-34) or Teriparatide using a cleavable fusion protein strategy was compared in two strains of E. coli. Materials and Methods: A cassette was designed and fully synthesized for prokaryotic expression of rhPTH using pET system. From 5’ to 3’, the cassette consisted of: Trx tag to increase the solubility of protein, His tag for purification and detection of protein, enterokinase site to cleave all fusion moieties, and an optimized gene code for Teriparatide corresponding to the amino acid sequence of hPTH. This cassette was cloned into pET32a vector. The vector was simultaneously transformed and expressed in two different E. coli strains. The ability of strains for expression of this recombinant pharmaceutical was compared. Early expression was confirmed by SDS-PAGE and Western Blotting. The soluble fusion protein was harvested and purified by immobilized affinity chromatography. Then the fusion moiety was released from Teriparatide by enterokinase digestion. Results: The fusion form of rhPTH was efficiently expressed in both E. coli strains. However, the percentage of the target protein to the total protein content in Rosetta-gami was more than its amount in BL21 (­60 % vs 25%).The fusion protein was highly purified with Ni-NTA column. Up to 18.5 mg/ml of pure fusion protein has been obtained from 1-liter Rosetta-gami strain of E. coli. The pure Teriparatide was released by enterokinase digestion. Conclusion: The pure rhPTH (1-34) produced here, could be the subject for biological activity and quality control assessments, and following formulation processing, it could be applied as a peptide drug in the treatment of Osteoporosis. &nbsp

BACKGROUND: Human monoclonal antibodies are important molecules in clinical research. Current Limitations of mAb technologies namely instability of immortalized B-cell line and probability of forming unusual VH-VL pairs in phage-display method led to mAbs technology based on single plasma cell called “SYMPLEX”. OBJECTIVE: In this method, cognate VH and VL fragments generated from individual antibody genes exactly the same as natural ones. METHODS: PBMCs of whole blood of an immunized candidate was used as a resource of rearranged Ab genes. Then flowcytometric screening was performed to isolate VH and VL from PBMCs. Various VH and VLκ were amplified by six pairs of primers. Overlap Extension PCR was accomplished to link VH and Vκ regions. ScFv inserted into T-vector and its sequence was determined and eventually analyzed by using blast analysis tools. RESULTS:Electrophoresis results indicated that VH and VL fragments were separately amplified by PCR with a length of about 400bp and linked through OE-PCR. Hence, ScFv, which was approximately 800bp in size, was constructed then sequencing and BLASTn results of the ScFv fragment consequently proved the accuracy. CONCLUSION: Results showed 88% similarity to available sequences in mentioned databank. ScFv was ultimately inserted into expression vector for producing recombinant human anti-tetanus mAb.

Background Over the last three decades, monoclonal antibodies have become powerful therapeutics and diagnostics tools. Progress in the antibody engineering, and the appearance of great selection technology such as phage display has made human antibodies production possible against antigens with high affinities. Objective The purpose of this study was to construct an immune antibody library from a vaccinated donor against tetanus toxin. Methods A blood sample was drawn from the donor who was vaccinated with tetanus toxoid. PBMC were isolated by using ficoll. After RNA extraction and cDNA synthesis. In order to amplify VH and VL regions, two uniplex PCRs were performed and put considering NcoI & HindIII, MluI & NotI restriction sites respectively for their cloning. These amplicons were cloned to pSEX81 vector and transformed to E. coli XL1blue strain. Eventually, recombinant plasmids were extracted and sequenced. Results The result showed acceptable similarity between antibody gene library nucleotide sequences and the antibody genes were deposited in this database. Conclusion In this study, the VH and VL genes human antibody library was constructed and confirmed by using DNA sequencing and alignment of sequences with blast database.