Physico-chemical and antimicrobial characteristics of novel biodegradable films based on gellan and carboxymethyl cellulose containing rosemary essential oil.

The purpose of the current investigation was to produce a novel functional composite biodegradable film by Gellan (Gla) and Carboxymethyl cellulose (CMC) biopolymers containing rosemary essential oils (REO) and evaluate their physicochemical and antimicrobial features. The film containing 5 % REO, due to its better mechanical properties (UTS = 13.44 ± 0.30 Mpa and SB = 21.14 ± 1.15 %) compared to other emulsified samples containing REO, was selected as the optimal film. Furthermore, it had less water vapor permeability (WVP = 6.60 ± 0.31 (g/mhPa) × 10-8) in comparison to control sample (8.21 ± 0.10 (g/mhPa) × 10-8) and the best color properties among the samples. The Scanning Electron Microscopy (SEM) images didn't show the phenomenon of agglomeration and point accumulation of REO. Also, 5 % of REO contributed to the increased compactness of the film in comparison to the film without the REO. Based on the results of Fourier-transform infrared spectroscopy (FTIR) spectra, no new chemical bonds were created by adding REO to the biopolymer substrate, and the REO was well dispersed and distributed among the Gla-CMC chains throughout the film substrate. Adding 5 % REO showed antioxidant effects. Considering the antimicrobial tests, all films containing REO had antimicrobial effects against the Staphylococcus aureus, Escherichia coli, Salmonella typhimurium, and Pseudomonas fluorescens bacterial strains.

The Caucasian Whortleberry Extract/Myrtle Essential Oil Loaded Active Films: Physicochemical Properties and Effects on Quality Parameters of Wrapped Turkey Breast Meat.

In this research work, the effects of myrtle essential oil (MEO) and Caucasian whortleberry extract (CWE) as natural additives were investigated on mechanical, physico-mechanical and antimicrobial properties of gellan/polyvinyl alcohol (G/PVA) film. Then, optimal blend active films were used for the wrapping of turkey breast meat stored at low temperature (4 ± 1 °C) for 15 days and chemical and sensory properties of wrapped meats were evaluated. The addition of MEO and CWE decreased tensile strength and increased the strain at the break of the films (p ≤ 0.05). Additionally, with increasing the amount of MEO and CWE, the permeability to water vapor (WVP) and the moisture content (MC) of the films decreased (p ≤ 0.05). MIC test showed that MEO and CWE were effective against S. aureus, E. coli, S. typhimurium, and P. fluorescens. at the concentrations of 5-6 and 15-17 mg/mL, respectively. Different microbiological, chemical, and sensory tests indicated that active films significantly enhanced the shelf life of turkey breast meat (p ≤ 0.05). Therefore, based on our finding in this study, the use of these active and biodegradable packagings can be effective and useful for protecting the microbial and sensory quality of turkey breast meat.

Rapid Isolation of Gastric Adenocarcinoma Cancer Stem Cells as a Target for Autologous Dendritic Cell-Based Immunotherapy.

Background: Gastric cancer (GC) is a malignancy cause associated with a high death rate in the world. Cancer stem cells (CSCs) are a rare immortal subpopulation of cells within tumors with characteristics of the ability to self-renew, initiate tumor, and differentiate into defined progenies as well as and high resistance to conventional therapies. Objectives: Despite the use of surgery and chemotherapy for GC therapy, there are no efficient therapeutic protocols for it to date. Therefore, rapid isolation of CSCs in order to therapeutic targets, especially immunotherapy is very important. Materials and Methods: Cancerous cell suspension isolated from patients with GC was cultured in the serum-free medium containing EGF, bFGF, LIF, and heparin under non-adherent culture conditions to generate spheres. Expression of mRNA level stemness transcription factors (OCT4, SOX2, SALL4, and Cripto-1), CD44 variable isoforms (CD44s, CD44v3, CD44v6, CD44V8-10) of spheroid-forming single cells compared with gastric normal tissue cells using real time PCR and molecules of CD44, CD54, and EpCAM as gastric CSC markers, and stemness factor Oct4 using flow cytometry, as well as tumorgenicity using subcutaneous injection of sphere-forming cells to nude mice were investigated. Results: Few cancerous cells isolated from patients with GC were able to generate three-dimensional spheroid colonies in the serum-free medium containing EGF, bFGF, LIF, and heparin under non-adherent culture conditions, and form xenograft tumors in immunodeficient nude mice after subcutaneous injection. Spheroid-forming single cells upregulated stemness transcription factors OCT4, SOX2, SALL4, and Cripto-1 that are associated with pluripotency and self-renewal and CD44 isoforms (CD44s, CD44v3, CD44v6, CD44V8-10) compared with gastric normal tissue cells. Finally, molecules of CD44, CD54, and EpCAM as gastric CSC markers and stemness factor Oct4 were expressed in sphere-forming cells. Conclusion: We suggested that the sphere formation and tumorigenicity assays are two procedures, leading to the rapid isolation of cancer cells with certain stem-like properties in order to target CSCs using autologous dendritic cell therapy, especially in patients with advanced disease.

S100A12 is not expressed in rodents: Transgenic mouse model is needed.

I read with interest the article authored by Zhang et al. (2020) who examined the role of S100A12 in the pathogenesis of sepsis-induced ARDS (acute respiratory distress syndrome). They used wild-type (WT) mice to measure the expression of S100A12 after induction of sepsis in mice. Based on the fact that S100A12 is not present in the murine genome, the interpretation of the findings could be misleading. Therefore, I discuss the use of S100A12 transgenic mouse models to examine S100A12 expression in mice.

Establishment and characterization of chemotherapy-enriched sphere-forming cells with stemness phenotypes as a new cell line (BAG50) of gastric carcinoma.

Gastric cancer is a malignancy with a high mortality rate worldwide. Cancer stem cells (CSCs) are a small subpopulation of tumor cells that possess the tumor-initiating ability, self-renewal capacity, and high resistance to conventional therapies. Due to the diversity and complexity of human tumors, new cell lines are urgently needed to supply clinically and physiologically relevant cancer models. Here, we report establishing a novel cell line (BAG50) with stemness properties. Chemotherapy-enriched sphere-forming cells with CSC properties isolated from a patient with GC were cultured in a serum-containing medium and passaged for up to 51 passages. The colony-forming ability and tumor-forming capacity of BAG50 cells were evaluated in vitro and in vivo. mRNA upregulation of stemness-related transcriptional factors using real-time PCR as well as expression of CSC markers using flow cytometry was investigated. Finally, STR profiling and chromosome studies were performed. BAG50 cells formed floating spheroid colonies in a serum-free medium. Subcutaneous injection of these cells generated xenograft tumors in nude mice. Pluripotency markers (SOX-2, OCT4, and Cripto-1) in them were upregulated compared with normal gastric cells. The majority of them expressed CSC markers of CD44, CD54, and EpCAM, and stemness marker of oct-4. STR profiling showed a unique DNA fingerprint. Karyotype also demonstrated multiple aneuploidies and chromosomal translocations. We suggested that the highly tumorigenic BAG50 cell line with stem cell-like phenotypes may provide a valuable in vitro tool to support new diagnostic, prognostic, and predictive biomarkers as well as the development of more effective treatment strategies.

Selectively targeting cancer stem cells: Current and novel therapeutic strategies and approaches in the effective eradication of cancer.

Cancer stem cells (CSCs) are a subgroup of cells in malignant cancers, which possess self-renewal capacity, tumor-initiating capability, and pluripotency, as well as being responsible for tumor maintenance, metastasis, relapse, and chemoresistance. The treatment modalities previously established for cancer included surgery, chemotherapy, and radiotherapy. The majority of tumor cells of non-CSCs could be eradicated using conventional chemotherapy and radiotherapy. Therefore, novel and promising therapeutic strategies that selectively target CSCs are of great importance. In this review, we described different therapeutic strategies such as immunotherapy, metabolism-based therapeutic strategies, and additional potential therapeutic approaches (targeting microRNAs [miRNAs], histone deacetylase, and DNA methyl transferase) against CSCs. Taken together, due to the inefficiency of anticancer single therapies, targeting CSCs through their metabolism and using immunotherapy and miRNAs besides classical chemo- and radiotherapy may exert better therapeutic effects.

Kinetics Analysis and Susceptibility Coefficient of the Pathogenic Bacteria by Titanium Dioxide and Zinc Oxide Nanoparticles.

Purpose: The increase of bacterial resistance to common antibacterial agents is one of the major problems of health care systems and hospital infection control programs. In this study, antimicrobial activity of titanium dioxide (TiO2 ) and zinc oxide (ZnO) nanoparticles (NPs) was investigated against E. coli, Salmonella enteritidis, Listeria monocytogenes, and Staphylococcus aureus pathogenic bacteria by determining sensitivity coefficient and kinetics of bacterial death. Methods: Antimicrobial tests were performed with ~106 CFU/mL of each bacterium at baseline. At first, minimum inhibitory concentration (MIC) was concluded by the dilution method and then, death kinetic and susceptibility coefficient of NPs suspensions was determined at 0 to 360 min. treatment time. Results: The results of this study revealed that, the highest susceptibility was observed for L. monocytogenes (Z=0.025 mL/µg) to TiO2 NPs, whereas the lowest susceptibility was obtained in the reaction of ZnO NPs with S. enteritidis (Z=0.0033 mL/µg). The process of bacterial death in NPs suspension was assumed to follow first-degree kinetic and the survival ratio of bacteria decreased by the increase in treatment time. An increase in the concentration of NPs was seen to enhance the bactericidal action. Conclusion: Results showed that L. monocytogenes had higher sensitivity compared to S. enteritidis. The results of this study also demonstrated that TiO2 NPs have a strong antimicrobial effect in comparison with ZnO NPs and it could be employed to aid the control of pathogenic bacteria.

Nanoparticles and Zeolites: Antibacterial Effects and their Mechanism against Pathogens.

Nowadays, distribution and microorganism resistance against antimicrobial compounds have caused crucial food safety problems. Hence, nanotechnology and zeolite are recognized as new approaches to manage this problem due to their inherent antimicrobial activity. Different studies have confirmed antimicrobial effects of Nano particles (NPs) (metal and metal oxide) and zeolite, by using various techniques to determine antimicrobial mechanism. This review includes an overview of research with the results of studies about antimicrobial mechanisms of nanoparticles and zeolite. Many researches have shown that type, particle size and shape of NPs and zeolite are important factors showing antimicrobial effectiveness. The use of NPs and zeolite as antimicrobial components especially in food technology and medical application can be considered as prominent strategies to overcome pathogenic microorganisms. Nevertheless, further studies are required to minimize the possible toxicity of NPs in order to apply suitable alternatives for disinfectants and antibacterial agents in food applications.

Induction of T cell-mediated immune response by dendritic cells pulsed with mRNA of sphere-forming cells isolated from patients with gastric cancer.

Gastric cancer (GC) as the third most common cause of cancer-associated mortality worldwide is one of the cancers with very high heterogeneity. Cancer stem cells (CSCs) as a small subset of cancer cells in solid tumors with the self-renewal, differentiation and tumorigenic ability are responsible for tumor initiation, progression, recurrence, metastasis, and resistance to current treatments. Therefore, eradication of CSCs is very vital to cure cancer. Here, we first isolated and identified sphere-forming cells in tumor tissue from four GC patients and then analyzed T cell responses induced by monocyte-derived dendritic cells (DCs) loaded with total mRNA of sphere-forming cells in terms of interferon-gamma (IFN-γ) gene expression and specific cytotoxicity. Spheroid colonies were formed in serum-free media. Sphere-forming cells dissociated from tumorspheres heterogeneously expressed CD44, CD54, and epithelial cell adhesion molecule (EpCAM) markers and generated one tumor in nude mice. These results demonstrated that gastric CSCs were enriched in tumorspheres. Cytokine-matured DCs loaded with mRNA of sphere-forming cells were able to induce IFN-γ gene expression in T-lymphocytes after a 12-day co-culture. mRNA level of IFN-γ gene in these lymphocytes was more highly expressed compared to stimulated T-lymphocytes by DCs transfected with normal tissue (6.4-9.39 folds). Cytotoxic activity of primed T-lymphocytes with antigens of sphere-forming cells was significantly higher than normal tissue antigens and mock DCs (P ≤ 0.0001). Taken together, DCs loaded with mRNA of sphere-forming cells that elicit effectively specific T cell-mediated immune responses in vitro, may be considered as a promising therapeutic vaccination in GC patients in future.

S100A12-CD36 axis: A novel player in the pathogenesis of atherosclerosis?

S100A12 is a member of the S100 family of EF-hand calcium-binding proteins and have a variety of intracellular and extracellular activities. It exerts its proinflammatory effects by binding to the receptor for advanced glycation end products (RAGE) and Toll-like receptor 4 (TLR4). CD36 is a class B scavenger receptor that acts as a fatty acid transporter. Both S100A12 and CD36 are implicated in vascular inflammation and atherosclerosis. It has recently been demonstrated that S100A12 binds with high affinity to CD36. On the other hand, RAGE and TLR4 play a key role in the regulation of CD36 expression. These observations point to the fact that S100A12 is an interesting molecular target for the development of therapeutics. This Cytokine stimulus will focus on the possible mechanisms of S100A12-CD36 axis in the pathogenesis of atherosclerosis.

CXCL12 and CXCR4 in the Peripheral Blood of Patients with Parkinson's Disease.

OBJECTIVE: The role of CXCL12 and its receptor CXCR4 has not been fully examined in Parkinson's disease (PD). The purpose of this study was to investigate the role of CXCL12/CXCR4 in the peripheral blood of patients with PD and healthy controls. METHODS: CXCL12 serum levels and CXCR4 mRNA levels were measured in 30 PD patients and 40 controls using ELISA and real-time PCR, respectively. RESULTS: CXCL12 serum levels were significantly higher in PD patients compared to controls (p < 0.0001). Moreover, CXCR4 expression in peripheral blood mononuclear cells (PBMC) of PD patients was significantly increased compared to controls (p < 0.0001). CONCLUSIONS: Our findings provide new information on the expression of CXCL12/CXCR4 in PD. CXCR4 expression in PBMC or CXCL12 serum levels may be potential biomarkers of inflammation in PD patients.

Isolation and identification of chemotherapy-enriched sphere-forming cells from a patient with gastric cancer.

Gastric cancer (GC) is the third and fifth cause of cancer-associated mortality for men and women throughout the world, respectively. Despite the use of surgery and chemotherapy for GC therapy, there are no efficient therapeutic protocols for it to date. Cancer stem cells (CSCs) due to their pivotal role in tumor initiation, growth, progression, invasion, distant metastasis, recurrence and resistance to anticancer drugs are very appealing targets for cancer therapies. Here, we isolated and identified CSCs from a chemotherapy-treated patient. Small subpopulation of dissociated cells after tissue digestion formed spheroid colonies in serum-free media under the non-adherent condition. These spheroid colonies differentiated into epithelial like cells in serum-containing medium. Few sphere-forming cells carried CD44 and CD54 markers overexpressed DLL4 that is responsible for tumor growth and angiogenesis. Subcutaneous injections of sphere-forming cells in different passages conferred tumorigenicity in nude mice. Sphere-forming cells upregulated CD44 polymorphisms CD44v3, -v6, and -v8 -10, stemness factors OCT4, SOX2, SALL4 and Cripto-1, self-renewal molecules IHh, Wnt, ß-catenin and BMI1, and epithelial mesenchymal transition (EMT) markers Twist1 and Snail1 in vitro and in vivo. Moreover, these cells similar to sphere-forming cells isolated from a chemotherapy-free patient expressed Oct-4 and ß-catenin proteins. However, the Twist1 protein was only expressed by sphere-forming cells derived from the chemotherapy-treated patient. Thus, these cells have all the characteristics of stationary and migratory CSCs, including tumorigenicity, self-renewal, pluripotency, invasion and metastasis. Taken together, targeting chemotherapy-enriched CSCs as chemo-resistance cells observed in GC patients can provide more effective therapeutic strategies compared to untreated patients.

Cytokine networks and their association with Helicobacter pylori infection in gastric carcinoma.

Cytokine networks as dynamic networks are pivotal aspects of tumor immunology, especially in gastric cancer (GC), in which infection, inflammation, and antitumor immunity are key elements of disease progression. In this review, we describe functional roles of well-known GC-modulatory cytokines, highlight the functions of cytokines with more recently described roles in GC, and emphasize the therapeutic potential of targeting the complex cytokine milieu. We also focus on the role of Helicobacter pylori (HP)-induced inflammation in GC and discuss how HP-induced chronic inflammation can lead to the induction of stem cell hyperplasia, morphological changes in gastric mucosa and GC development.

Toll like receptor 4: an important molecule in recognition and induction of appropriate immune responses against Chlamydia infection.

Chlamydia species are obligate intracellular pathogens causing different infectious diseases particularly asymptomatic genital infections and are also responsible for a wide range of complications. Previous studies showed that there are different immune responses to Chlamydia species and their infections are limited to some cases. Moreover, Chlamydia species are able to alter immune responses through modulating the expression of some immune system related molecules including cytokines. Toll like receptors (TLRs) belonge to pathogen recognition receptors (PRRs) and play vital roles in recognition of microbes and stimulation of appropriate immune responses. Therefore, it appears that TLRs may be considered as important sensors for recognition of Chlamydia and promotion of immune responses against these bacterial infections. Accordingly, TLR4 detects several microbial PAMPs such as bacterial lipopolysacharide (LPS) and subsequently activates transcription from pro-inflammatory cytokines in both MYD88 and TRIF pathways dependent manner. The purpose of this review is to provide the recent data about the status and major roles played by TLR4 in Chlamydia species recognition and promotion of immune responses against these infections and also the relationship between TLR4 activities and pathogenesis of Chlamydia infections.

S100A12: Friend or foe in pulmonary tuberculosis?

In humans, S100A12 (also named Calgranulin C and EN-RAGE) is mainly expressed and secreted by neutrophil granulocytes. Extracellular S100A12 is involved in innate immune responses against microorganisms and parasites. S100A12 is a ligand for the receptor for advanced glycation end products (RAGE), which is a cell surface receptor on macrophages, endothelium, and lymphocytes. In a recent study, Realegeno et al. showed that S100A12 exerts antimicrobial activity against Mycobacterium leprae in infected human macrophages. Recently, some interesting data on the antimicrobial activity of S100A12 have been reported. Proinflammatory role of S100A12 is supported by another newly found receptor, Toll-like receptor 4 (TLR4). These observations emphasize the importance of S100A12 for the development of potential therapeutic approaches to increase protective immunity or reduce immunopathogenesis.

Neutralizing human recombinant antibodies against herpes simplex virus type 1 glycoproteins B from a phage-displayed scFv antibody library.

The HSV-1 envelope glycoprotein B (gB) plays a critical role in virus entry into host cells. Neutralizing antibodies can therefore potentially prevent virus entry into target cells and cell-to-cell spread of infection. Our present study focused on the selection of neutralizing single-chain Fv (scFv) antibodies of a phage-displayed nonimmune human scFv antibody library against gB of HSV-1. To enrich specific scFvs, two phage antibodies were isolated against amino acid residues 31-43 derived from the N-terminal part of gB using panning technique. Two scFvs, scFv-gB1 and scFv-gB2, with frequencies of 45% and 20% were obtained from scFv clones after performing PCR and MvaI fingerprinting. In phage ELISA analysis, both gB1 and gB2 scFvs demonstrated high reactivity with the gB peptide. In the neutralization assay, scFv-gB1 and scFv-gB2 represented neutralizing effects of 55% and 59%, respectively. Upon further enhancement of the neutralizing effects of these antibodies, they can be considered as new potential alternatives in the treatment and prophylaxis of HSV-1 infections.

Isolation, identification, and characterization of cancer stem cells: A review.

Cancer stem cells (CSCs) or tumor-initiating cells (TICs) as a small subset of neoplastic cells are able to produce a tumor (tumorigenesis), maintain the population of tumorigenic cells (self-renewal), and generate the heterogeneous cells constructing the entire tumor (pluripotency). The research on stationary and circulating CSCs due to resistance to conventional therapies and inability in complete eradication of cancer is critical for developing novel therapeutic strategies for a more effective reduction in the risk of tumor metastasis and cancer recurrence. This review compiles information about different methods of detection and dissociation, side population, cellular markers, and establishment culture of CSCs, as well as characteristics of CSCs such as tumorigenicity, and signaling pathways associated with self-renewal and the capability of the same histological tumor regeneration in various cancers.