Anti-cancer Effects of a Chitosan Based Nanoformulation Expressing miR-340 on 4T1 Breast Cancer Cells.

MicroRNAs (miRNAs) have a crucial role in the regulation of gene expression in tumor development, invasion, and metastasis. Herein, miRNA-340 (miR-340) has been shown to play tumor suppressor activity in breast cancer (BC). However, the clinical applications of miRNAs request the development of safe and effective delivery systems capable of protecting nucleic acids from degradation. In this study, biodegradable chitosan nanoparticles incorporating miR-340 plasmid DNA (pDNA) (miR-340 CNPs) were synthesized and characterized. Then, the anti-tumor effects of miR-340 CNPs were investigated using 4T1 BCE cells. The spherical nanoparticles (NPs) with an appropriate mean diameter of around 266 ± 9.3 nm and zeta potential of +17 ± 1.8 mV were successfully prepared. The NPs showed good stability, high entrapment efficiency and a reasonable release behavior, meanwhile their high resistance against enzymatic degradation was verified. Furthermore, NPs demonstrated appropriate transfection efficiency and could induce apoptosis, so had toxicity in 4T1 BCE cells. Also, CD47 expression on the surface of cancer cells was significantly reduced after treatment with miR-340 CNPs. The results showed that miR-340 CNPs augmented the expression of P-27 in BC cells. Furthermore, miR-340 CNPs caused down-regulation of BRP-39 (breast regression protein-39) increasingly suggested as a prognostic biomarker for neoplastic diseases like BC. In conclusion, our data show that miR-340 CNPs can be considered as a promising new platform for BC gene therapy.

Study the effect of recombinant leukemia inhibitory factor on maintenance of pregnancy and frequency of regulatory T cells in abortion-prone mice.

Recurrent spontaneous abortion (RSA) can have a significant impact on a woman's quality of life. Understanding the mechanisms behind abortion is crucial for developing potential treatments. Among various models of abortion, the CBA/J(♀) × DBA/2J(♂) model stands out as the most extensively studied. This model reveals the influence of an altered immune system on resorption during pregnancy. The leukemia inhibitory factor (LIF) holds considerable importance as a secretory glycoprotein essential for successful implantation. Regulatory T cells (Tregs) have been found to produce high levels of LIF in both mice and humans. LIF plays a vital role in the development of Tregs by upregulating the expression of the Foxp3 transcription factor while downregulating the expression of RORγt. To investigate the impact of recombinant LIF (rLIF) on pregnancy maintenance and Treg cell frequency in abortion-prone (AP) mice, a specific recombinant protein was used in this study. The AP group consisted of CBA/J(♀) × DBA/2J(♂) mice, while the control group comprised CBA/J(♀) × BALB/c(♂) mice. Intraperitoneal injections of rLIF were administered to the AP group on the third day of pregnancy, and its effects on Treg cell frequency and pregnancy maintenance were examined during this period. Following rLIF injections on the fourteenth day of pregnancy, the expression of Foxp3 significantly increased in AP mice (p = 0.02,0.008). Additionally, AP mice injected with rLIF demonstrated a significant reduction in resorption rate (p = 0.01) and a notable increase in birth rate (p = 0.01,0.0005). These findings provide new insights into the potential benefits of LIF in treating RSA patients.

Detailed numerical evaluation of diffusion convection equation in layered reservoirs during tracer injection.

Characterization of heterogeneous reservoirs such as multilayered or fractured systems is an important issue in different disciplines such as hydrology, petroleum and geothermal systems. One of the popular methods that can be used for this purpose is tracer tests. Better understanding of the mechanisms of mass transfer (convection-diffusion process) is essential for having a proper test interpretation. In this study, the solutions of different scenarios of tracer flow in a pair of high and low-permeable layered reservoirs including convection and diffusion mechanisms are discussed. Although analytical solutions generally provided exact solutions, they involve several assumptions and might be hard to use for complex problems. As a result, numerical methods are selected for the investigation of different scenarios and addressing cases that are beyond access of analytical methods. In this study, several scenarios of considering diffusion and convection in low and high permeable zones and effective parameters on tracer concentration are investigated. According to the results of this study, the higher the porosity ratio of low to high permeable layer, the more time is needed to get the final concentration value. Also, by increasing the value of the dispersivity coefficient, the time needed to increase the concentration decreases. In other words, the sharp increase in concentration for lower times is seen in higher dispersivity values. The concentration profile variation is affected by Peclet number. The difference among concentration profiles in different cases is considerable, especially in low Peclet numbers where the diffusion mechanism is dominant. This behavior is more common in low permeable mediums such as multilayered tight or shale reservoirs.

Investigation of Expression Profile of Placenta-specific 1 (PLAC1) in Acute Myeloid and Lymphoid Leukemias.

Background: Placenta-specific 1 (PLAC1) is one of the cancer-testis-placenta antigens that has no expression in normal tissue except placenta trophoblast and testicular germ cells, but is overexpressed in a variety of solid tumors. There is a lack of studies on the expression of PLAC1 in leukemia. We investigated expression of PLAC1 in Acute Myeloid Leukemia (AML) and Acute Lymphoblastic Leukemia (ALL). Methods: In this study, we investigated expression pattern of PLAC1 gene in peripheral blood and bone marrow mononuclear cells of newly-diagnosed patients with AML (n=31) and ALL (n=31) using quantitative real-time PCR. Normal subjects (n=17) were considered as control. The PLAC1 protein expression in the samples were also detected using western blotting. Results: Our data demonstrated that PLAC1 transcripts had 2.7 and 2.9 fold-change increase in AML and ALL, respectively, compared to normal samples. PLAC1 transcript expression was totally negative in all studied normal subjects. Level of PLAC1 mRNA expression in ALL statistically increased compared to normal samples (p=0.038). However, relative mRNA expression of PLAC1 in AML was not significant in comparison to normal subjects (p=0.848). Furthermore, relative mRNA expression of PLAC1 in AML subtypes was not statistically significant (p=0.756). PLAC1 gene expression showed no difference in demographical clinical and para-clinical parameters. Western blotting confirmed expression of PLAC1 in the ALL and AML samples. Conclusion: Considering PLAC1 expression profile in acute leukemia, PLAC1 could be a potential marker in leukemia which needs complementary studies in the future.

Evaluation of Molecular-level Changes of Programmed Cell Death Ligand-1 after Radiation Therapy in a BALB/c CT26 Colorectal Mouse Tumor Model.

The effects of radiation therapy (RT) for cancer can be systemic and partially mediated by the immune system. However, radiation alone is unlikely to transform an immunosuppressive environment into an immunostimulatory one. Therefore, an effective combination of RT and immunotherapy may provide a new, more efficient treatment approach. Here, we investigated how the expression of programmed cell death-ligand 1 (PD-L1) in the tumor microenvironment varied in different RT regimens with the same biologically effective dose. In this study, female BALB/c mice inoculated with CT26 tumor cells were irradiated with 3 different RT regimens using the same BED of 40 gray (Gy). These included ablative RT (1*15 Gy), hypo-fractionated RT (2*10 Gy), and conventional (Hyper-fractionated) RT (10*3 Gy). PD-L1 expression was analyzed with immunohistochemical staining on days 2 and 20 and when the size of tumors had reached 2 cm2 after RT. All treated groups expressed PD-L1, but the group receiving single ablative high-dose RT showed higher expression compared to the other groups. No significant differences in PD-L1 expression were observed at different times in the same group. These findings showed that different regimens of RT have different effects on the TME, so a combination of RT and immune checkpoint blockade could be clinically used in cancer patients.

Evaluation of the anti-tumor effects of an anti-Human Epidermal growth factor receptor 2 (HER2) monoclonal antibody in combination with CD11b+/Gr-1+ myeloid cells depletion using a recombinant peptibody in 4 T1-HER2 tumor model.

INTRODUCTION: Clinical efficacy of Human Epidermal growth factor Receptor 2 (HER2) targeted strategies is limited due to impaired anti-tumor responses negatively regulated by immunosuppressive cells. We thus, investigated the inhibitory effects of an anti-HER2 monoclonal antibody (1 T0 mAb) in combination with CD11b+/Gr-1+ myeloid cells depletion in 4 T1-HER2 tumor model. METHODS: BALB/c mice were challenged with human HER2-expressing 4 T1 murine breast cancer cell line. A week post tumor challenge, each mouse received 50 µg of a myeloid cells specific peptibody every other day, or 10 mg/kg of 1 T0 mAb two times a week, and their combination for two weeks. The treatments effect on tumor growth was measured by calculating tumor size. Also, the frequencies of CD11b+/Gr-1+ cells and T lymphocytes were measured by flow cytometry. RESULTS: Peptibody treated mice indicated tumor regression and 40 % of the mice eradicated their primary tumors. The peptibody was capable to deplete notably splenic CD11b+/Gr-1+ cells as well as intratumoral CD11b+/Gr-1+ cells (P < 0.0001) and led to an increased number of tumor infiltrating CD8+ T cells (3.3 folds) and also that of resident tumor draining lymph nodes (TDLNs) (3 folds). Combination of peptibody and 1 T0 mAb resulted in enhanced expansion of tumor infiltrating CD4 + and CD8+ T cells which was associated with tumor eradication in 60 % of the mice. CONCLUSIONS: Peptibody is able to deplete CD11b+/Gr-1+ cells and increase anti-tumoral effects of the 1 T0 mAb in tumor eradication. Thus, this myeloid population have critical roles in development of tumors and their depletion is associated with induction of anti-tumoral responses.

Proteome Analysis of Adult Acute Lymphoblastic Leukemia by Two-dimensional Blue Native/Sodium Dodecyl Sulfate Gel Electrophoresis.

Background: Despite the significant progress in the treatment of Acute Lymphoblastic Leukemia (ALL) in children, it still remains as one of the most challenging malignancies in adults. Identification of new biomarkers may improve the management of adult ALL. Proteins expressed on the cell surface can be considered as disease-associated biomarkers with potential for diagnosis and targeted therapies. Thus, membrane proteome studies give essential information about the disease-related biomarkers. Methods: We applied 2-dimensional blue-native SDS-PAGE technique followed by MALDI-TOF/TOF-mass spectrometry to study the cell membrane proteome of peripheral blood mononuclear cells of adult B-ALL patients in comparison to that of the healthy controls. Results: Sixty seven differentially expressed protein spots were detected, among them 52 proteins were found to be up-regulated but the other 15 proteins were down-regulated in B-ALL. Five differentially expressed proteins, involved in energy metabolism pathways, were detected in B-ALL patients compared to the healthy control group. Conclusion: Differentially expressed proteins provide an insight into the molecular biology of B-ALL. Further studies must be done to confirm our data to be considered as potential targets for detection and treatment of B-ALL.

Myeloid-derived suppressor cells (MDSCs) depletion by cabozantinib improves the efficacy of anti-HER2 antibody-based immunotherapy in a 4T1-HER2 murine breast cancer model.

BACKGROUND: Clinical trials using Cabozantinib have shown promising results in metastatic breast cancer. This efficacy mainly results from removing and/or polarization of tumor-promoting myeloid cells. Nevertheless, whether such myeloid-derived suppressor cells (MDSCs) depletion can be used to improve the efficacy of anti-HER2 antibodies in early breast cancer has not been defined yet. METHODS: BALB/c mice were inoculated with 4T1 and 4T1-HER2 murine tumor cell lines, and after 7 days, the mice were divided into different groups. Cabozantinib was orally administrated for 15 consecutive days, and anti-HER2 monoclonal antibody (mAb) 1 T0 was intraperitoneally injected twice a week. Tumor size was measured every other day. RESULTS: Our findings indicated that Cabozantinib combined with anti-HER2 mAb dramatically reduced tumor growth and increased tumor rejection (p = 0.0001). Flow cytometry analysis showed MDSC population decreased in TME, lymph nodes, and spleens by roughly 20%, 0.8%, and 35%, respectively. Myeloid suppressive phenotype was altered through inhibition of the expression of immunosuppressive factor Arg-1. Cytokine profiling of different groups indicated that the level of INF-γ was approximately two times higher than that in the control group, and IL-17 increased compared to the control group. However, IL-4 level was significantly reduced in the groups treated with Cabozantinib. These could bring about a 10% increase in CD8+ infiltration into the tumor bed and activation of tumor-draining lymph nodes and splenic T-lymphocytes. CONCLUSION: Collectively, our data provide pre-clinical evidence for using Cabozantinib to reshape the primary TME, which can enhance the effectiveness of anti-HER2 mAb immunotherapy in primary breast cancer.

Immunologic deviations in recurrent spontaneous abortion mouse model.

BACKGROUND: Recurrent spontaneous abortion (RSA) or recurrent pregnancy loss (RPL) is an abnormality that has a great impact on women's quality of life. RSA is defined as at least three unexplained abortions (without prior live birth) occurring before the 20th week of pregnancy. AIM: The present review attempts to discuss immunologic deviations in mouse models of RSA. CONTENT: The mating of DBA/2J males with CBA/J female mice has provided specialists with a homologous model of RSA. Much of the research using the CBA/J × DBA/2J mouse model has shown immune system alteration results in rejection. The link between RSA and the immune system suggests new approaches to prevent RSA from an immunological perspective. Rejection in this model is linked with the changed immune system during pregnancy, including change in Th1/Th2 ratio and defects in T and NK cells function, and so forth. IMPLICATIONS: The use of animal models prone to RSA can help a lot to solve the remained mysteries. This study reviews the existing knowledge of immune system roles in the RSA mouse models.

Investigation of Durability of SARS-CoV-2-specific IgG and IgM Antibodies in Recovered COVID-19 Patients: A Prospective Study.

Background: Evidence on seroconversion profile of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infected patients is limited. We mainly aimed to evaluate seroconversion and persistence of virus-specific antibodies in patients infected by coronavirus disease 2019 (COVID-19). Methods: This prospective study was conducted on 118 patients with COVID-19 presentations admitted to three hospitals in Iran and recovered from the disease, during April and May 2020. Presence of COVID-19 was confirmed by Polymerase Chain Reaction (PCR) testing on nasopharyngeal swabs. Serum samples were collected at different time points, including 0-5, 6-15, 16-25, 26-35, and 36-95 days of clinical symptom onset. For measurement of SARS-CoV-2-specific IgG and IgM antibody titers, Iran's Food and Drug Administration-approved SARS-CoV-2 ELISA kits were used. Results: Serologic assay revealed that 37.3% of patients (n=44) were positive for IgM at 0-5 days interval after clinical symptom onset. This rate was 60.2% (n=71) for IgG. There were increasing IgM and IgG seroconversion rates during first 25 days of clinical symptom onset, but seropositivity started to decrease thereafter, which was more evident for IgM (17.9%) than IgG (58.9%) at the 36-95 days post symptoms appearance. In other words, it was found that 83.6% of IgM-positive and 32.9% of IgG-positive patients in the first month of clinical symptom onset became seronegative in the third month of clinical symptom onset. Conclusion: The findings demonstrated that antibody responses to SARS-CoV-2 infection were developed in recovered COVID-19 patients; however, some of them were seronegative three months after onset of relevant symptoms. Furthermore, the stability of anti-SARS-CoV-2 antibodies could also correct our expectations from COVID-19 vaccination responses.

Cloning, expression and characterization of a peptibody to deplete myeloid derived suppressor cells in a murine mammary carcinoma model.

BACKGROUND: Myeloid derived suppressor cells (MDSCs) are an immature heterogeneous population of myeloid lineage that attenuate the anti-tumor immune responses. Depletion of MDSCs has been shown to improve efficacy of cancer immunotherapeutic approaches. Here, we expressed and characterized a peptibody which had previously been defined by phage display technique capable of recognizing and depleting murine MDSCs. MATERIALS AND METHODS: Using splicing by overlap extension (SOE) PCR, the coding sequence of the MDSC binding peptide and linker were synthesized and then ligated into a home-made expression plasmid containing mouse IgG2a Fc. The peptibody construct was transfected into CHO-K1 cells by lipofectamine 3000 reagent and the resulting fusion protein was purified with protein G column and subsequently characterized by ELISA, SDS-PAGE and immunoblotting. The binding profile of the peptibody to splenic MDSCs and its MDSC depletion ability were then tested by flow cytometry. RESULTS: The purified peptibody appeared as a 70 KDa band in Western blot. It could bind to 98.8% of splenic CD11b+/Gr-1+ MDSCs. In addition, the intratumoral MDSCs were significantly depleted after peptibody treatment compared to their PBS-treated negative control counterparts (P < 0.05). CONCLUSION: In this study, a peptibody capable of depleting intratumoral MDSCs, was successfully expressed and purified. Our results imply that it could be considered as a potential tool for research on cancer immunotherapy.

Tim-3 and PD-1 blocking cannot restore the functional properties of natural killer cells in early clinical stages of chronic lymphocytic leukemia: An in vitro study.

Background: Programmed death-1 (PD-1) and T-cell immunoglobulin- and mucin-domain-containing molecule-3 (Tim-3) are two major immune checkpoint receptors expressed on immune cells and their expression is related to the exhaustion phenotype. In the present in vitro study, blocking of PD-1 and Tim-3 molecules was performed on isolated natural killer (NK) cells from patients with chronic lymphocytic leukemia (CLL) to restore their functional properties. Materials and Methods: NK cells fraction was positively isolated from fresh peripheral blood of 18 CLL patients, treated with anti-PD-1 and anti-Tim-3 blocking monoclonal antibodies and co-cultured with K562 target cells to evaluate their apoptosis induction by Annexin V-PI method. Blocked NK cells were also incubated with anti-CD107a antibody to assess their degranulation properties by flow cytometry. The level of secreted tumor node factor-alpha (TNF-α) and interferon-gamma (IFN-γ) by NK cells was also measured by ELISA. Results: Our results showed similar functional properties in terms of degranulation and apoptosis of K562 target cells by isolated NK cells from CLL patients in PD-1/Tim-3 blocked and control groups. It was also shown that blocking of PD-1 and Tim-3 could not improve the production of pro-inflammatory TNF-α and IFN-γ cytokines by isolated NK cells from CLL patients. Conclusion: Altogether, our results indicated that pretreatment of NK cells with anti-PD-1 and anti-Tim-3 blocking antibodies in CLL patients at early clinical stages cannot improve their functional properties. Besides many other malignancies, the application of checkpoint inhibitors in CLL needs more investigations and complementary studies.

Antioxidant nutrients can increase high-dose Methotrexate efficacy in 4T1 breast tumor Model: An experimental study on Vitamin E Succinate and Methyl-selenic acid.

BACKGROUND: We aimed to evaluate the anti-cancer and immune system enhancing properties of Vitamin E succinate (VES) and methylselenic acid (MSA) administration on 4T1 breast tumor model under high-dose methotrexate (HDMTX) therapy and folinic acid (FA) rescue. METHODS: Thirty six 4T1 mammary carcinoma bearing mice were randomly divided into six groups: control (untreated; n = 6), treatment-1 (T1 group; HDMTX; n = 6), T2 (T1 + FA; n = 6), T3 (T2 + MSA; n = 6), T4 (T2 + VES; n = 6) and T5 (T3 + VES; n = 6). On day 21 of the study, all surviving mice were sacrificed and primary tumors and peripheral tissues were examined for histological and gene expression assays. The expression of GATA Binding Protein-3 (GATA3), forkhead box-P3 (FOXP3), T-bet and Retinoic acid receptor-related orphan receptor γt (RORγt) were evaluated in tumors and spleens. Also, vascular endothelial growth factor-A (VEGF-A) and UL16-Binding Protein 1 (ULBP-1) expression were evaluated in tumors. RESULTS: The control, T4 and T5 groups were able to complete the entire 21-day study period. Also, significant tumor shrinkage was occurred in T4 group (P < 0.05). Suppression of splenic FOXP3 and GATA3 were observed in the mice receiving T4 and T5 regimens. Also, induction of tumoral FOXP3 and GATA3 were achieved in the T4 and T5 groups, respectively (P < 0.05). No metastasis occurred in T4 receiving group; while, lung and liver metastasis were observed in T5 group. CONCLUSION: In this study, high and fixed dose of MTX was used. Further studies are needed to optimize MTX dose along with FA, VES and MSA.

The engineered probiotics for the treatment of chronic diseases: A systematic review.

Engineered probiotics (EPs) are a group of probiotics whose proteome is manipulated by biotechnological techniques. EPs have attracted a lot of attention in recent researches for preventing and treating chronic diseases. The current study has been conducted to provide an overview regarding the EPs application in the treatment of chronic disease by a comprehensive systematic review of the published articles up to January 2022. To retrieve the related publications, three databases (PubMed/MEDLINE, Web of Sciences, and Scopus) were searched systematically. Finally, all human (n = 2) and animal (n = 37) studies were included. The included articles evaluated the effects of EPs on treatment of arthritis (n = 3), cancer (n = 2), autoimmune encephalomyelitis (EAE; n = 6), Parkinson disease (PD; n = 1), Alzheimer diseases (AD; n = 1), colitis (n = 11), celiac disease (n = 1), diabetes (n = 8) and cardiovascular disease (CVD; n = 6). Induction of oral tolerance (OT) is the most important mechanism of EPs action in the treatment of chronic disease. Providing oral vaccine and bioactive compounds are the other mechanisms of EPs action. PRACTICAL APPLICATIONS: The current systematic review gathered evidence about the application of EPs in the treatment of chronic diseases. Evidence suggests that EPs have very broad and potent effects in the treatment of chronic and even genetic diseases.

Evaluation the reactivity of a peptide-based monoclonal antibody derived from OmpA with drug resistant pulsotypes of Acinetobacter baumannii as a potential therapeutic approach.

BACKGROUND: Acinetobacter baumannii is an opportunistic and antibiotic-resistant pathogen that predominantly causes nosocomial infections. There is urgent need for development nonantibiotic-based treatment strategies. We developed a novel monoclonal antibody (mAb) against a peptide of conserved outer membrane protein A (OmpA) and evaluated its reactivity with different pulsotypes of A. baumannii. METHODS: Peptide derived from A. baumannii OmpA was conjugated to keyhole limpet hemocyanin and injected into BALB/c mice. Splenocytes of immunized mice were fused with SP2/0 myeloma cells followed by selection of antibody-producing hybridoma cells. After screening of different hybridoma colonies by ELISA, one monoclone was selected as 3F10-C9 and the antibody was tested for reaction with five different Acinetobacter pulsotypes that were resistant to carbapenem antibiotics. The affinity constant was measured by ELISA. The ELISA, western blotting, indirect immunofluorescence (IFA), and in vitro opsonophagocytosis assays were used to evaluate the reactivity of generated mAb. RESULTS: The anti-OmpA antibody reacted with the immunizing peptide and had a high affinity (1.94 × 10-9 M) for its antigen in the ELISA. Specific binding of mAb to OmpA was confirmed in Western blot. IFA assays revealed that mAb recognized specific OmpA on the pulsotypes. Opsonophagocytosis assays showed that the mAb increased the bactericidal activity of macrophage cells. The antibody function was higher in the presence of serum complement. CONCLUSIONS: The peptide-based mAb demonstrated optimal performance in laboratory experiments which may be appropriate in investigation on OmpA in Acinetobacter pathogenesis and development of passive immunization as a novel therapeutic approach.

Functionalized SPION immobilized on graphene-oxide: Anticancer and antiviral study.

The progressive and fatal outbreak of some diseases such as cancer and coronavirus necessitates using advanced materials to bring such devastating illnesses under control. In this study, graphene oxide (GO) is decorated by superparamagnetic iron oxide nanoparticles (SPION) (GO/SPION) as well as polyethylene glycol functionalized SPION (GO/SPION@PEG), and chitosan functionalized SPION (GO/SPION@CS). Field emission scanning electron microscopic (FESEM) images show the formation of high density uniformly distributed SPION nanoparticles on the surface of GO sheets. The structural and chemical composition of nanostructures is confirmed by X-ray diffraction and Fourier transform infrared spectroscopy. The saturation magnetization of GO/SPION, GO/SPION@PEG and GO- SPION@CS are found to be 20, 19 and 8 emu/g using vibrating sample magnetometer. Specific absorption rate (SAR) values of 305, 283, and 199 W/g and corresponding intrinsic loss power (ILP) values of 9.4, 8.7, and 6.2 nHm2kg-1 are achieved for GO/SPION, GO/SPION@PEG and GO/SPION@CS, respectively. The In vitro cytotoxicity assay indicates higher than 70% cell viability for all nanostructures at 100, 300, and 500 ppm after 24 and 72 h. Additionally, cancerous cell (EJ138 human bladder carcinoma) ablation is observed using functionalized GO/SPION under applied magnetic field. More than 50% cancerous cell death has been achieved for GO/SPION@PEG at 300 ppm concentration. Furthermore, Surrogate virus neutralization test is applied to investigate neutralizing property of the synthesized nanostructures through analysis of SARS-CoV-2 receptor-binding domain and human angiotensin-converting enzyme 2 binding. The highest level of SARS-CoV-2 virus inhibition is related to GO/SPION@CS (86%) due to the synergistic exploitation of GO and chitosan. Thus, GO/SPION and GO/SPION@PEG with higher SAR and ILP values could be beneficial for cancer treatment, while GO/SPION@CS with higher virus suppression has potential to use against coronaviruses. Thus, the developed nanocomposites have a potential in the efficient treatment of cancer and coronavirus.

Virulence genes expression profiling of different Shigella flexneri serotypes in response to sub-inhibitory concentrations of azithromycin and ciprofloxacin.

BACKGROUND: Shigellosis is a self-limiting disease that antibiotic therapy could decrease its complications and duration. However, sublethal levels of antibiotics, may lead to alteration in disease state, besides its role in the emergence of resistant variants. To understand this link, we investigated diversity of Shigella serogroups in children with diarrhea, diversity of S. flexneri serotypes, cytotoxic potential, resistance patterns to antibiotics, and alteration in transcriptional expression of main virulence genes in response to sub-inhibitory concentrations of azithromycin and ciprofloxacin. RESULTS: The most frequently isolated serogroups were S. sonnei (70.3%), followed by S. flexneri (29.1%) and S. boydii (0.6%). Ten serotypes were characterized among the S. flexneri isolates, including 2b, 1b, 2a, 1c, 4a, 3a, 3b, 6 and X and/or Xv. Antimicrobial susceptibility testing showed low frequency of multi-drug resistance phenotype among S. flexneri isolates with minimum inhibitory concentrations (MIC) of 0.5-64 and 0.25-8 µg/mL for azithromycin and ciprofloxacin, respectively. Gene expression analysis showed upregulation of icsA in serotype 4a after exposure with azithromycin, whereas other genes in the VirF pathway were downregulated, and downregulation of virB in serotypes 2a and 3a after exposure with ciprofloxacin, while upregulation of noted genes was detected. CONCLUSIONS: Alteration in transcription of key virulence genes of S. flexneri serotypes was shown in response to sublethal concentration of antibiotics. The detected incongruency in the extent of gene transcription proposed that diverse regulatory pathways are possibly mediating response to sub-MIC concentrations of antibiotics in S. flexneri.

The Association between Consumption of Dairy-Originated Digestion Resistant and Bioactive Peptides and Breast Cancer Risk: A Case-Control Study.

Bioactive peptides (BPs) content of dairy products is suggested to be a significant ingredient for reducing breast cancer (BC) risk. There is no observational study regarding the correlation between BPs and the risk of chronic disease because BPs' content of food items has not been evaluated in any study. The goal of the current study was to assess the association of dairy-originated BPs with BC risk. One hundred thirty-four women with BC and 267 cancer-free controls were selected from referral hospitals in Tehran, Iran. The development of an in-silico model for estimation of the bioactive and digestion-resistant peptides content of dairy products was done in our previous research. The risk assessment for BPs and BC association was performed across the tertiles of the peptide's intake. Odds ratios (OR) were calculated by logistic regression. The negative association of all bioactive and digestion-resistant peptides except for peptides with high hydrophilicity and low bioactivity was seen in all models. In PR-negative subjects only the association of total dairy intake (OR: 0.61; 95% CI: 0.26-1.45; P for trend: 0.276), peptides with low bioactivity (OR: 0.40; 95% CI: 0.16-1.02; P for trend: 0.0.052), antidiabetic peptides (OR: 0.42; 95% CI: 0.17-1.05; P for trend: 0.0.062) and di-peptides (OR: 0.42; 95% CI: 0.17-1.05; P for trend: 0.0.062) were not significant in the final model. Also, no significant association between ER-negative subjects and total dairy intake (OR: 0.41; 95% CI: 0.16-1.07; P for trend: 0.0.068) was noted. Our findings deduced that milk-derived BPs negatively associate with the risk of ER/PR/HER2 negative BC among Iranian women.Supplemental data for this article is available online at https://doi.org/10.1080/01635581.2021.2009884.

Correlations and diagnostic tools for metabolic syndrome (MetS) and chronic obstructive pulmonary disease (COPD).

BACKGROUND: Regarding the importance of obesity in patients with chronic obstructive pulmonary disease (COPD), we aimed to evaluate of correlation between metabolic syndrome (MetS) and COPD. METHODS: In this cross-sectional study, 96 patients with COPD were evaluated. This study was conducted in 2016-2018. The severity of COPD was determined by Global Initiative for Chronic Obstructive Lung Disease (GOLD) 2017 criteria. We investigated the correlations between MetS with COPD and possible diagnostic tools. RESULTS: Of all COPD patients, 86.5% had MetS, and the means of waist circumference, fasting blood glucose, systolic and diastolic blood pressure, body mass index, and triglyceride in patients with MetS were significantly higher than the patients without MetS (P < 0.05). We showed that forced expiratory volume in 1 second (FEV1) with a 37% cutoff had 92.8% and 69.2% sensitivity and specificity, respectively (area of the curve: 0.51, 0.31-0.71). CONCLUSION: MetS is prevalent among COPD and FEV1 could be considered as important diagnostic tool for COPD.

Galectin-9 inhibits cell proliferation and induces apoptosis in Jurkat and KE-37 acute lymphoblastic leukemia cell lines via caspase-3 activation.

BACKGROUND AND PURPOSE: Acute lymphoblastic leukemia (ALL) is a type of cancer of blood and bone marrow characterized by abnormal proliferation of lymphoid progenitor cells. Galectin-9 is a tandem-repeat type galectin expressed in various tumor cells. It seems that the connection between galectin-9 and T cell immunoglobulin mucin-3 receptor acts as a negative regulator of cancer cells proliferation. EXPERIMENTAL APPROACH: In this research, the effects of galectin-9 were investigated using MTS cell proliferation colorimetric, colony-forming, annexin V-FITC/PI, and caspase-3 assays in the Jurkat and KE-37 cell lines of ALL. Furthermore, the western blotting technique was used to evaluate the levels of apoptotic proteins such as Bax and Bcl-2 in these cell lines. FINDINGS/RESULTS: Our results indicated that galectin-9 can considerably reduce the cell growth and colony formation ability of both Jurkat and KE-37 cell lines in a concentration-dependent manner. Besides, galectin-9 induced apoptosis in a concentration-dependent manner in ALL cells by a mechanism associated with Bax/Bcl-2 expression and activation of the caspase-3 activation. CONCLUSION AND IMPLICATIONS: Galectin-9 inhibited the growth and proliferation of cell lines with increased programmed cell death, therefore it can be considered as a potential factor in the progression of ALL therapeutics that needs more research in this context.