Design and Production of Arginine Deiminase-Azurin Recombinant Fusion Protein from Pseudomonas aeruginosa and its Confirmation by Western Blot

Background: Pseudomonas aeruginosa is a common Gram-negative, rod-shaped bacterium that has a unique genome that allowed the bacteria to produce various enzymes and proteins. Azurin and arginine deiminase are low molecular weight proteins that produced by P. aeruginosa. These proteins can be used alone or in combination together in order to become effective in cancer therapy or inhibiting of metastasis. This study aimed to design, express and purify the Azurin and Arginine Deiminease recombinant fusion protein.Materials and Methods: The sequences of Azurin and arginine deiminase from P. aeruginosa were studied for synthesis in a pET28a vector. The recombinant plasmid was transfected into the E.coli BL-21 strain and expression was induced by isopropyl-β-D-thio galactopyranoside (IPTG). The fusion protein expression was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Purification of the recombinant product was performed by the Ni-NTA chromatography column, obtained product analysis with SDS-PAGE and Western blot technique.Results: Cloning was confirmed by observing bands from the enzyme digestion. The protein band with a molecular weight of 65 kDa on the SDS-PAGE gel was an indication of the correct expression of the protein. The single-band of this recombinant protein was confirmed by the western blot technique.Conclusion: In this study, due to the successful production of arginine deiminase-azurin fusion protein, and considering the separate anti-cancer properties of these compounds, which have been reported in previous studies, it is suggested that immunological assessments and effects of this fusion protein in different cancerous cell line be investigated

Cytotoxic Activity of Nisin on Human Cancer Cell Lines: A Systematic Review

Cancer is one of the leading causes of mortality and morbidity worldwide. Nisin consists of various and uncommon amino acids as an antimicrobial protein synthesized by the Lactobacilli genus. The current systematic review aimed to evaluate the anticancer activity of Nisin, an antibacterial peptide, on different human cancer cell lines. We searched PubMed/Medline and Embase databases to detect the studies addressing the cytotoxic activity of Nisin on human cancer cell lines. Our study was conducted following the "PRISMA" guideline. Of 202 potentially relevant articles, 15 studies met the inclusion criteria and were included for further analysis. The results revealed that Nisin has different levels of anticancer activity on human cancer cell lines. The outcomes of our review indicate that some cancer cell lines, such as cell skin carcinoma (A431), Melanoma cells (A375), and colorectal cancer cell lines (LS180), are strongly affected by the anticancer properties of Nisin. In contrast, the anticancer effect of Nisin on others like Human promyelocytic leukemia (HL60) is lower. Nisin shows significant anticancer effects in different cancer cell lines. Utilizing Nisin simultaneously with other antitumor agents can enhance its anticancer features and efficacy. Further studies, especially in vivo assay and clinical trials, are recommended to achieve more accurate results in this field

Investigation of the Potential Presence of Porphyromonas gingivalis in Esophageal Squamous Cell Carcinoma (ESCC) Paraffin Embedded Tissue Samples

Background and Aim: Esophageal cancer is the eighth most common cancer and the sixth leading cause of cancer death worldwide Evidence suggests that there is a link between bacterial infection and malignancy. There are few studies on the prevalence of Porphyromonas gingivalis in esophageal squamous cell carcinoma (ESCC), so this study aimed to investigate the possible presence of this bacterium in ESCC tissue samples.Materials and Methods: In this study, 34 esophageal squamous cell carcinoma samples were collected to evaluate the potential presence of Porphyromonas gingivalis. After extracting the DNA, the polymerase chain reaction (PCR) technique was used to detect the presence of the bacterium molecularly.Results: The age range of the study population was 26 to 90 years, with a mean age of 63 years. Most tissue samples come from stage I cancer (73.5%). Based on the molecular analysis, no P. gingivalis was detected in any biopsy specimensConclusion: P. gingivalis infection and ESCC were not correlated based on the current in this study. Likely, the use of fresh samples, more accurate diagnostic methods, geographic differences, and larger sample sizes all contribute to the differences in results between related research, which can be clarified through large-scale studies

Investigation of Antibacterial Effect of Cuminum cyminum and Carum carvi against Streptococcus mutans and Streptococcus pyogenes

Background: Streptococcus mutans is major cause of tooth decay. In addition, there is some report about effects of Streptococcus pyogenes on odontogenic infections. Herbal drugs is now administrating for treatment of some disease. The aim of this study was to evaluate the antibacterial effect of Cuminum cyminum and Carum carvi against Streptococcus mutans and Streptococcus pyogenes.Materials and Methods: In this study, standard strains of Streptococcus mutans and Streptococcus pyogenes were used, and ethanolic extract was prepared in the microbiological laboratory of Shahid Beheshti University of Medical Sciences. The experiment was repeated 6 times and chlorohexine 2% was used as a control.Results: The best effect of Carum carvi was on Streptococcus mutans, whereas on Streptococcus pyogenes both extracts of Cuminum cyminum and Carum carvi showed the same effect.Conclusion: This study showed, due to the fact that the Cuminum cyminum and Carum carvi extracts exhibited excellent antibacterial properties, they could be a suitable candidate in compounds such as mouthwash, toothpastes and Gums. Therefore, could be used in the prevention and treatment of future oral and dental diseases

Genetic Diversity Analysis of Methicillin-resistant Staphylococcus aureus Strains Isolated from Intensive Care Unit in Iran

Objectives: Staphylococcus aureus has emerged as a major public health concern. It is a common pathogen in high-risk hospital intensive care units (ICUs). We analyzed the molecular characteristics on the SCCmec and spa genes of S. aureus isolates gathered from ICUs. The antibiotic resistance patterns and carriage of resistance and virulence determinants were also identified. Methods: In this cross-sectional study, 84 non-duplicated S. aureus strains isolated from ICU patients in were genotyped using SCCmec and spa typing. The Kirby-Bauer disk diffusion and micro-broth dilution methods were used to determine resistance patterns. Virulence and resistance gene profiling were also determined using the polymerase chain reaction technique. Results: All isolates were methicillin-resistant S. aureus and belonged to seven spa types: t388 (36.9%), t852 (14.3%), t924 (13.1%), t790 (11.9%), t064 (10.7%), t037 (9.5%), and t084 (3.6%). They differed in the carriage of resistance and toxin genes. The most common SCCmec type was III detected in 50 isolates (59.5%), followed by type IV in 34 isolates (40.5%). The pvl gene was detected in 14.3% (n = 12) of isolates, of which 66.7% (n = 8) belonged to t852 and 33.3% (n = 4) belonged to t790. Among the tested strains, 9.5% (n = 8) carried the mupA gene and belonged to the t064 spa type. Conclusions: The data revealed a high resistance rate to antibiotics, which could be a threat to ICU patients. It is necessary to detect antimicrobial resistance and resistance and toxin-encoding of gene profiles in different molecular types

Skin Manifestations in COVID-19 Patients: Are They Indicators for Disease Severity? A Systematic Review

Introduction: Until now, there are several reports on cutaneous manifestations in COVID-19 patients. However, the link between skin manifestations and the severity of the disease remains debatable. We conducted a systematic review to evaluate the temporal relationship between different types of skin lesions and the severity of COVID-19. Methods: A systematic search was conducted for relevant studies published between January and July 2020 using Pubmed/Medline, Embase, and Web of knowledge. The following keywords were used: SARS-CoV-2 or COVID-19 or new coronavirus or Wuhan Coronavirus or coronavirus disease 2019 and skin disease or skin manifestation or cutaneous manifestation. Results: Out of 381 articles, 47 meet the inclusion criteria and a total of 1,847 patients with confirmed COVID-19 were examined. The overall frequency of cutaneous manifestations in COVID-19 patients was 5.95%. The maculopapular rash was the main reported skin involvement (37.3%) commonly occurred in middle-aged females with intermediate severity of the disease. Forty-eight percentage of the patients had a mild, 32% a moderate, and 20% a severe COVID-19 disease. The mild disease was mainly correlated with chilblain-like and urticaria-like lesions and patients with vascular lesions experienced a more severe disease. Seventy-two percentage of patients with chilblain-like lesions improved without any medication. The overall mortality rate was 4.5%. Patients with vascular lesions had the highest mortality rate (18.2%) and patients with urticaria-like lesions had the lowest mortality rate (2.2%). Conclusion: The mere occurrence of skin manifestations in COVID-19 patients is not an indicator for the disease severity, and it highly depends on the type of skin lesions. Chilblain-like and vascular lesions are the ends of a spectrum in which from chilblain-like to vascular lesions, the severity of the disease increases, and the patient\u27s prognosis worsens. Those with vascular lesions should also be considered as high-priority patients for further medical care

Pulmonary Manifestations of SARS Co V 2 Infection in Mild/Severe Patients

The coronavirus disease 2019 (COVID-19) caused viral pneumonia in Wuhan City in China in December of 2019. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) primarily targets the lungs with severe hypoxia, which usually results in death. COVID-19 is highly heterogeneous regarding severity, clinical phenotype, and more importantly, global dispersal. The respiratory system in all aspects such as respiratory airways, endothelium of pulmonary vessels, conducting airways, the alveoli, neuromuscular breathing structure, and pulmonary circulation are affected by this virus. A comprehensive concept of the source and dynamic action of the SARS-CoV-2 and the possible causes of heterogeneity in COVID-19 is required for predicting and managing the illness in acute and chronic stages of the pulmonary sign

Investigation of the Potential Presence of Porphyromonas gingivalis in Esophageal Squamous Cell Carcinoma (ESCC) Paraffin Embedded Tissue Samples

Background and Aim: Esophageal cancer is the eighth most common cancer and the sixth leading cause of cancer death worldwide Evidence suggests that there is a link between bacterial infection and malignancy. There are few studies on the prevalence of Porphyromonas gingivalis in esophageal squamous cell carcinoma (ESCC), so this study aimed to investigate the possible presence of this bacterium in ESCC tissue samples. Materials and Methods: In this study, 34 esophageal squamous cell carcinoma samples were collected to evaluate the potential presence of Porphyromonas gingivalis. After extracting the DNA, the polymerase chain reaction (PCR) technique was used to detect the presence of the bacterium molecularly. Results: The age range of the study population was 26 to 90 years, with a mean age of 63 years. Most tissue samples come from stage I cancer (73.5%). Based on the molecular analysis, no P. gingivalis was detected in any biopsy specimens Conclusion: P. gingivalis infection and ESCC were not correlated based on the current in this study. Likely, the use of fresh samples, more accurate diagnostic methods, geographic differences, and larger sample sizes all contribute to the differences in results between related research, which can be clarified through large-scale studies

Diagnostic Accuracy of Pyrazinamide Susceptibility Testing in Mycobacterium tuberculosis: A Systematic Review with Meta-Analysis

Introduction: Pyrazinamide (PZA) susceptibility testing plays a critical role in determining the appropriate treatment regimens for multidrug-resistant tuberculosis. We conducted a systematic review and meta-analysis to evaluate the diagnostic accuracy of sequencing PZA susceptibility tests against culture-based susceptibility testing methods as the reference standard. Methods: We searched the MEDLINE/PubMed, Embase, and Web of Science databases for the relevant records. The QUADAS-2 tool was used to assess the quality of the studies. Diagnostic accuracy measures (i.e., sensitivity and specificity) were pooled with a random-effects model. All statistical analyses were performed with Meta-DiSc (version 1.4, Cochrane Colloquium, Barcelona, Spain), STATA (version 14, Stata Corporation, College Station, TX), and RevMan (version 5.3, The Nordic Cochrane Centre, the Cochrane Collaboration, Copenhagen, Denmark) software. Results: A total of 72 articles, published between 2000 and 2019, comprising data for 8,701 isolates of Mycobacterium tuberculosis were included in the final analysis. The pooled sensitivity and specificity of the PZA sequencing test against all reference tests (the combination of BACTEC mycobacteria growth indicator tube 960 (MGIT 960), BACTEC 460, and proportion method) were 87% (95% CI: 85–88) and 94.7% (95% CI: 94–95). The positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio, and the area under the curve estimates were found to be 12.0 (95% CI: 9.0–16.0), 0.17 (95% CI: 0.13–0.21), 106 (95% CI: 71–158), and 96%, respectively. Deek's test result indicated a low likelihood for publication bias (p = 0.01). Conclusions: Our analysis indicated that PZA sequencing may be used in combination with conventional tests due to the advantage of the time to result and in scenarios where culture tests are not feasible. Further work to improve molecular tests would benefit from the availability of standardized reference standards and improvements to the methodology

Cloning and Expression of Truncated Chlamydial Major Outer Membrane Protein in E.coli: A Miniature Step Forward

Administration of Prophylactic vaccines still is the cutting-edge of prevention, control and elimination of infections. Among the common causes of sexually transmitted diseases, chlamydia trachomatis is an important socioeconomic burden worldwide which impairs human genital tract function, resulting in severe complications like urethritis, pelvic inflammatory disease (PID), getting ease of HIV transmission, playing cofactor role in human papilloma virus (HPV)- induced cervical neoplasia, and unfortunately infertility. Because of all difficulties, vaccination is considered to be the most economical and reliable way to escape from unrestrained exacerbation states of infection than any other prevalence program. But despite much advancement in the field of veterinary, the dream has yet to be realized. Among a number of potential candidates, the major outer membrane protein (MOMP) is a vanguard of subunit vaccines. In this attempt, MOMP217 gene fragment cloned in PET28b+ and expressed in E.coli. The protein expression confirmed by SDS-PAGE. These findings demonstrate that, considering the antigenicity prediction and due to the existence of potential epitopic region, rMOMP217 could be a potential capable peptide which can represent as an alternative to whole MOMP