Development of Low-Serum Culture Media for the in Vitro Cultivation of Theileria annulata S15 Cell Line.

Background: The production of bovine theileriosis vaccine involves in vitro cultivation of Theileria annulata schizont-infected cell lines. Fetal bovine serum (FBS) is commonly used in animal cell culture, including the Theileria cell line. However, we aimed to reduce the amount of serum needed for cell culture by modifying the Stoker culture medium with supplements such as excretion factor and serum substitutes. Methods: To evaluate the effectiveness of these modifications, techniques such as cell counting, cell viability assays, and genomic analysis were employed in the Parasitic Vaccines Production Department of Razi institute of Iran, from 2020 to 2022. Statistical analysis was used to compare the results of different experimental conditions. Results: The three experimental media were as effective as the commonly used 10% Stoker medium in supporting the growth and viability of cells. Conclusion: The significant reduction in the required amount of serum and the remarkable cell growth achieved by using defined serum replacements for the production of cell culture media is a significant step towards the preparation of a proper cell culture medium for the production of bovine Theileriosis vaccine.

The effect of EGCG/tyrosol-loaded chitosan/lecithin nanoparticles on hyperglycemia and hepatic function in streptozotocin-induced diabetic mice.

We aimed to study the potential of epigallocatechin-3-gallate/tyrosol-loaded chitosan/lecithin nanoparticles (EGCG/tyrosol-loaded C/L NPs) in streptozotocin-induced type 2 diabetes mellitus (T2DM) mice. The EGCG/tyrosol-loaded C/L NPs were created using the self-assembly method. Dynamic light scattering, Field Emission Scanning Electron Microscopy, and Fourier transform infrared spectroscopy were utilized to characterize the nanoparticle. Furthermore, in streptozotocin-induced T2DM mice, treatment with EGCG/tyrosol-loaded C/L NPs on fasting blood sugar levels, the expression of PCK1 and G6Pase, and IL-1ß in the liver, liver glutathione content, nanoparticle toxicity on liver cells, and liver reactive oxygen species were measured. Our findings showed that EGCG/tyrosol-loaded C/L NPs had a uniform size distribution, and encapsulation efficiencies of 84 % and 89.1 % for tyrosol and EGCG, respectively. The nanoparticles inhibited PANC-1 cells without affecting normal HFF cells. Furthermore, EGCG/tyrosol-loaded C/L NP treatment reduced fasting blood sugar levels, elevated hepatic glutathione levels, enhanced liver cell viability, and decreased reactive oxygen species levels in diabetic mice. The expression of gluconeogenesis-related genes (PCK1 and G6 Pase) and the inflammatory gene IL-1ß was downregulated by EGCG/tyrosol-loaded C/L NPs. In conclusion, the EGCG/tyrosol-loaded C/L NPs reduced hyperglycemia, oxidative stress, and inflammation in diabetic mice. These findings suggest that EGCG/tyrosol-loaded C/L NPs could be a promising therapeutic option for type 2 diabetes management.

Safety and efficacy of RCP recombinant spike protein covid-19 vaccine compared to Sinopharm BBIBP: A phase III, non-inferiority trial.

Background: We conducted a phase III, non-inferiority trial comparing safety and efficacy of RCP recombinant spike protein Covid-19 vaccine to BBIBP (Sinopharm). Methods: Adult Iranian population received RCP or BBIBP in a randomized, double blind and an additional non-randomized open labeled trial arms. Eligible participants signed a written informed consent and received two intramuscular injections three weeks apart. In the randomized arm, an intranasal dose of vaccine or adjuvant-only preparation were given to the RCP and BBIBP recipients at day 51 respectively. Participants were actively followed for up to 4 months for safety and efficacy outcomes. Primary outcome was PCR + symptomatic Covid-19 disease two weeks after the second dose. The non-inferiority margin was 10% of reported BBIBP vaccine efficacy (HR = 1.36). Results: We recruited 23,110 participants (7224 in the randomized and 15,886 in the non-randomized arm). We observed 604 primary outcome events during 4 months of active follow-up including 121 and 133 in the randomized and 157 and 193 cases in the non-randomized arms among recipients of RCP and BBIBP respectively. Adjusted hazard ratios for the primary outcome in those receiving RCP compared with BBIBP interval were 0.91 (0.71-1.16) and 0.62 (0.49-0.77) in the randomized and non-randomized arms respectively. The upper boundary of 99.1% confidence interval of HR = 0.91 (0.67-1.22) remained below the margin of non-inferiority in the randomized arm after observing the early stopping rules using O'Brien Fleming method. Conclusion: Our study showed that the RCP efficacy is non-inferior and its safety profile is comparable to the BBIBP.

Immunogenicity and safety of RAZI recombinant spike protein vaccine (RCP) as a booster dose after priming with BBIBP-CorV: a parallel two groups, randomized, double blind trial.

BACKGROUND: The immunity induced by primary vaccination is effective against COVID-19; however, booster vaccines are needed to maintain vaccine-induced immunity and improve protection against emerging variants. Heterologous boosting is believed to result in more robust immune responses. This study investigated the safety and immunogenicity of the Razi Cov Pars vaccine (RCP) as a heterologous booster dose in people primed with Beijing Bio-Institute of Biological Products Coronavirus Vaccine (BBIBP-CorV). METHODS: We conducted a randomized, double-blind, active-controlled trial in adults aged 18 and over primarily vaccinated with BBIBP-CorV, an inactivated SARS-CoV-2 vaccine. Eligible participants were randomly assigned (1:1) to receive a booster dose of RCP or BBIBP-CorV vaccines. The primary outcome was neutralizing antibody activity measured by a conventional virus neutralization test (cVNT). The secondary efficacy outcomes included specific IgG antibodies against SARS-CoV-2 spike (S1 and receptor-binding domain, RBD) antigens and cell-mediated immunity. We measured humoral antibody responses at 2 weeks (in all participants) and 3 and 6 months (a subgroup of 101 participants) after the booster dose injection. The secondary safety outcomes were solicited and unsolicited immediate, local, and systemic adverse reactions. RESULTS: We recruited 483 eligible participants between December 7, 2021, and January 13, 2022. The mean age was 51.9 years, and 68.1% were men. Neutralizing antibody titers increased about 3 (geometric mean fold increase, GMFI = 2.77, 95% CI 2.26-3.39) and 21 (GMFI = 21.51, 95% CI 16.35-28.32) times compared to the baseline in the BBIBP-CorV and the RCP vaccine groups. Geometric mean ratios (GMR) and 95% CI for serum neutralizing antibody titers for RCP compared with BBIBP-CorV on days 14, 90, and 180 were 6.81 (5.32-8.72), 1.77 (1.15-2.72), and 2.37 (1.62-3.47) respectively. We observed a similar pattern for specific antibody responses against S1 and RBD. We detected a rise in gamma interferon (IFN-γ), tumor necrosis factor (TNF-α), and interleukin 2 (IL-2) following stimulation with S antigen, particularly in the RCP group, and the flow cytometry examination showed an increase in the percentage of CD3 + /CD8 + lymphocytes. RCP and BBIBP-CorV had similar safety profiles; we identified no vaccine-related or unrelated deaths. CONCLUSIONS: BBIBP-CorV and RCP vaccines as booster doses are safe and provide a strong immune response that is more robust when the RCP vaccine is used. Heterologous vaccines are preferred as booster doses. TRIAL REGISTRATION: This study was registered with the Iranian Registry of Clinical Trial at www.irct.ir , IRCT20201214049709N4. Registered 29 November 2021.

Antimicrobial and antibiofilm properties of selenium-chitosan-loaded salicylic acid nanoparticles for the removal of emerging contaminants from bacterial pathogens.

In this study salicylic acid loaded containing selenium nanoparticles was synthesized and called SA@CS-Se NPs. the chitosan was used as a natural stabilizer during the synthesis process. Fourier transforms infrared spectroscopy (FTIR), Powder X-ray diffraction (XRD), field emission electron microscopy (FESEM), and transmission electron microscopy (TEM) were used to describe the physicochemical characteristics of the SA@CS-Se NPs. The PXRD examination revealed that the grain size was around 31.9 nm. TEM and FESEM techniques showed the spherical shape of SA@CS-Se NPs. Additionally, the analysis of experiments showed that SA@CS-Se NPs have antibacterial properties against 4 ATCC bacteria; So that with concentrations of 75, 125, 150, and 100 µg/ml, it inhibited the biofilm formation of Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus respectively. Also, at the concentration of 300 µg/ml, it removed 22.76, 23.2, 10.62, and 18.08% biofilm caused by E. coli, P. aeruginosa, B. subtilis, and S. aureus respectively. The synthesized SA@CS-Se NPs may find an application to reduce the unsafe influence of pathogenic microbes and, hence, eliminate microbial contamination.

The folate-linked chitosan-coated Kaempferol/HSA nano-transporters (FCKH-NTs) as the selective apoptotic inducer in human MCF-7 breast cancer cell line.

BACKGROUND: Kaempferol, the natural bioactive flavonoid, has been utilized as an efficient anti-breast cancer compound. In the current study, the Kaempferol's cellular uptake and its aqueous solubility were improved by using human serum albumin (HSA) as the Kaempferol adjuvant and encapsulating it with the folate-linked chitosan polymer to evaluate the apoptotic, activity of the novel-formulated Kaempferol in human MCF-7 breast cancer cells. METHODS: The folate-linked chitosan-coated Kaempferol/HSA nano-transporters (FCKH-NTs) were synthesized and characterized using FTIR, FESEM, DLS, and Zeta potential analysis. The nano-transporters' selective cytotoxicity was studied by applying an MTT assay on the cancerous MCF-7 cells compared with normal HFF cell lines. Cell death type determination was determined by analyzing the expression of apoptotic (BAX and Cas-8) and anti-apoptotic genes (BCL2 and NF-κB). The FCKH-NTs apoptotic activity was verified by studying the flow cytometry and AO/PI staining results. RESULT: The 126-nm FCKH-NTs (PDI = 0.282) selectively induced apoptotic death in human MCF-7 breast cancer cells by up-regulating the BAX, Nf- κB, and Cas-8 gene expression. The apoptotic activity of FCKH-NTs was verified by detecting the SubG1-arrested cancer cells and increased apoptotic bodies in AO/PI staining images. CONCLUSION: The FCKH-NTs exhibited a selective-cytotoxic impact on human MCF-7 breast cancer cells compared with normal HFF cells, which can be due to the folate receptor-mediated endocytosis mechanism of the nano-transporters. Therefore, the FCKH-NTs have the potential to be used as a selective anti-breast cancer compound.

Phase II, Safety and Immunogenicity of RAZI Cov Pars (RCP) SARS Cov-2 Vaccine in Adults Aged 18-70 Years; A Randomized, Double-Blind Clinical Trial.

BACKGROUND: This study explores the safety and immunogenicity of the Razi-Cov-Pars (RCP) SARS Cov-2 recombinant spike protein vaccine. METHOD: In a randomized, double-blind, placebo-controlled trial, adults aged 18-70 were randomly allocated to receive selected 10 µg/200 µl vaccine strengths or placebo (adjuvant). It included two intramuscular injections at days 0 and 21, followed by an intranasal dose at day 51. Immediate and delayed solicited local and systemic adverse reactions after each dose up to a week, and specific IgG antibodies against SARS Cov-2 spike antigens two weeks after the 2nd dose were assessed as primary outcomes. Secondary safety outcomes were abnormal laboratory findings and medically attended adverse events (MAAE) over six months follow up. Secondary immunogenicity outcomes were neutralizing antibody activity and cell-mediated immune response. RESULT: Between May 27th and July 15th, 2021, 500 participants were enrolled. Participants' mean (SD) age was 37.8 (9.0), and 67.0 % were male. No immediate adverse reaction was observed following the intervention. All solicited local and systemic adverse events were moderate (Grade I-II). Specific IgG antibody response against S antigen in the vaccine group was 5.28 times (95 %CI: 4.02-6.94) the placebo group with a 75 % seroconversion rate. During six months of follow-up, 8 SAEs were reported, unrelated to the study intervention. The participants sustained their acquired humoral responses at the end of the sixth month. The vaccine predominantly resulted in T-helper 1 cell-mediated immunity, CD8+ cytotoxic T-cell increase, and no increase in inflammatory IL-6 cytokine. CONCLUSION: RCP vaccine is safe and creates strong and durable humoral and cellular immunity. TRIAL REGISTRATION: (IRCT20201214049709N2).

Synthesis and its characterisation of selenium/silver/chitosan and cellular toxicity against liver carcinoma cells studies.

Liver cancer is one of the most common lethal malignancy in the world. To treat liver cancer, new cure options are crucial. The use of natural substances along nanosciences may provide healing with lower toxicity and a smaller amount of side properties. In this research, The three-component selenium-silver-chitosan nanocomposite (Se-Ag-CS NCs) were synthesised with the help of ultrasound in a stepwise manner. The as-synthesised Se-Ag-CS NCs were characterised accordingly by applying powder X-Ray diffraction (PXRD), Fourier transforms infrared spectroscopy (FTIR), field emission scanning electron microscopy (FESEM), energy dispersive x-ray analysis (EDX), transmission electron microscopy (TEM), dynamic light scattering (DLS) and potential. The PXRD demonstrated that the NCs were synthesised successfully and the grain sizes of 27.3 were obtained. The FESEM and TEM analyses have shown the NCs have a nano-sized structure with spherical and rod-like morphologies in a coating of CS. The DLS analysis also revealed that NCs were synthesised in nanoscale particles. The NCs' surface charge was also positive due to the presence of chitosan. Different concentrations of NCs (0, 0.125, 0.250, 0.500, and 1 mg/ml) were tested at different times (24, 48, and 72 h) to measure cytotoxicity against liver cancer cells. The results showed at a concentration of 1 mg/mL in 72 h, the most toxicity effects were applied to liver cancer cells. Moreover, the results indicated NCs can inhibit the growth of cancer cells in a dose-dependent manner, while the toxicity of nanocomposite on normal cells was less. It is important to create nanocomposites derived from natural polymers as a new strategy in cancer treatment that can fight cancer cells while having low toxicity for normal cells. Therefore, the present results can be considered in improving cancer-fighting methods.

Immunogenicity and Safety of a Combined Intramuscular/Intranasal Recombinant Spike Protein COVID-19 Vaccine (RCP) in Healthy Adults Aged 18 to 55 Years Old: A Randomized, Double-Blind, Placebo-Controlled, Phase I Trial.

Objectives: This study aimed to determine the safety and immunogenicity of a combined intramuscular/intranasal recombinant spike protein COVID-19 vaccine (RCP). Methods: We conducted a randomized, double-blind, placebo-controlled, phase I trial. Three vaccine strengths were compared with an adjuvant-only preparation. It included two intramuscular and a third intranasal dose. Eligible participants were followed for adverse reactions. Specific IgG, secretory IgA, neutralizing antibodies, and cell-mediated immunity were assessed. Results: A total of 153 participants were enrolled (13 sentinels, 120 randomized, 20 non-randomized open-labeled for IgA assessment). No related serious adverse event was observed. The geometric mean ratios (GMRs) and 95% CI for serum neutralizing antibodies compared with placebo two weeks after the second injection were 5.82 (1.46-23.13), 11.12 (2.74-45.09), and 20.70 (5.05-84.76) in 5, 10, and 20 µg vaccine groups, respectively. The GMR for anti-RBD IgA in mucosal fluid two weeks after the intranasal dose was 23.27 (21.27-25.45) in the 10 µg vaccine group. The humoral responses were sustained for up to five months. All vaccine strengths indicated a strong T-helper 1 response. Conclusion: RCP is safe and creates strong and durable humoral and cellular immunity and good mucosal immune response in its 10 µg /200 µL vaccine strengths. Trial registration: IRCT20201214049709N1.

Evaluation of Serum Selenium and Copper Levels with Inflammatory Cytokines and Indices of Oxidative Stress in Type 2 Diabetes.

Type 2 diabetes mellitus (T2DM) is a metabolic and multifactorial disease in which inflammatory markers, oxidative stress, and certain trace elements seem to have an essential role. This study investigated the relationship between serum selenium and copper level with inflammatory cytokines and oxidative stress in T2DM.In this case-control study, 30 patients with T2DM and 30 healthy individuals were selected. Serum levels of copper and selenium were measured by atomic absorption spectrometry, and TNF-α and IL-6 and oxidative stress markers were measured by ELISA. The SPSS v.22 was used for data analysis and the significance level is less than 5%.The mean age of patients was 52.9 ± 10.4 years, and the control group was 48.5 ± 10.4 years. In this study, 53.3% were female, and 46.7% were male. The levels of BMI (p = 0.002), systolic pressure (p = 0.034), insulin, selenium, malondialdehyde, and glutathione peroxidase (p = 0.0001; each), insulin resistance, copper, and superoxide dismutase, IL6, and TNF-α (p = 0.001; each) in T2DM were significantly higher than the control group. While levels of lipid profile, uric acid, creatinine, and diastolic pressure were not significantly different between the two groups. Selenium and copper are related to insulin resistance, and their increasing levels are associated with increased levels of markers of oxidative stress and inflammatory cytokines (p < 0.05).Increased levels of copper and selenium are associated with T2DM and this increase is also associated with increased levels of TNF-α, IL-6, and oxidative stress in T2DM. Therefore, controlling these markers can lead us to control this disease better.

ZnO/CeO2 Nanocomposites: Metal-Organic Framework-Mediated Synthesis, Characterization, and Estimation of Cellular Toxicity toward Liver Cancer Cells.

The Zinc-doped cerium oxide nanocomposite (ZnO/CeO2 NC) was synthesized using a metal-organic framework as a precursor through the combustion method. It was characterized by powder X-ray diffraction (PXRD), Fourier transform infrared spectroscopy (FTIR), field emission electron microscopy (FESEM), energy dispersive analysis (EDX), transmission electron microscopy (TEM), dynamic light scattering (DLS), and ξ-potential. The PXRD demonstrated the successful synthesis of ZnO/CeO2 NC with a crystallite size of 31.9 nm. FESEM and TEM images displayed hexagonal and spherical morphologies, and the solid-phase size was 65.03 ± 30.86 nm for ZnO/CeO2 NCs. DLS, TEM, and FESEM showed that the NCs have a high tendency for agglomeration/aggregation in both aqueous media and solid phase. The anticancer attributes of ZnO/CeO2 NC were investigated against Liver cancer cells (HepG2), which showed inhibition of cancer cell growth on a concentration-dependent gradient. The cell toxicity effects of ZnO/CeO2 nanocomposites were also studied toward NIH-3T3, in which the data displayed the lower toxicity of NC compared to the HepG2 cell line.

A Combination of MALDI-TOF MS Proteomics and Species-Unique Biomarkers' Discovery for Rapid Screening of Brucellosis.

Brucellosis is considered to be a zoonotic infection with a predominant incidence in most parts of Iran that may even simply involve diagnostic laboratory personnel. In the present study, we apply matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for rapid and reliable discrimination of Brucella abortus and Brucella melitensis, based on proteomic mass patterns from chemically treated whole-cell analyses. Biomarkers of the low molecular weight proteome in the MALDI-TOF MS spectra were assigned to conserved ribosomal and structural protein families that were found in genome assemblies of B. abortus and B. melitensis in the NCBI database. Significant protein mass signals successfully mapped to ribosomal proteins and structural proteins, such as integration host factor subunit alpha, cold-shock proteins, HU family DNA-binding protein, ATP synthase subunit C, and GNAT family N-acetyltransferase, with specific biomarker peaks that have been identified for each virulent and vaccine strain. Web-accessible bioinformatics algorithms, with a robust data analysis workflow, followed by ribosomal and structural protein mapping, significantly enhanced the reliable assignment of key proteins and accurate identification of Brucella species. Furthermore, clinical samples were analyzed to confirm the most dominant protein biomarker candidates and their relevance for the identifications of B. melitensis and B. abortus. With proper optimization, we envision that the presented MALDI-TOF MS proteomics analyses, coupled with special usage of bioinformatics, could be used as a cost-efficient strategy for the diagnostics of brucellosis and introduce a reliable identification protocol for species of dangerous bacteria.

Prognostic value of HIF-1α in digestive system malignancies: evidence from a systematic review and meta-analysis.

Aim: This meta-analysis aimed to evaluate the association of HIF-1α expression with clinicopathological features and overall survival (OS) of patients with digestive system malignancies. Background: Numerous studies have demonstrated that hypoxia-inducible factor-1α (HIF-1α) is abnormally expressed in various solid tumors. However, the clinicopathological features and prognostic value of HIF-1α expression in patients with digestive system malignancies remain controversial. Methods: A literature search in PubMed, Web of Science, and Scopus databases was performed to identify all relevant studies published in English until 15 October 2020. The pooled effect was calculated to evaluate the association between HIF-1α expression and clinicopathological features and overall survival in cancer patients. Pooled odds ratios (ORs) or hazard ratios (HRs) with a 95% confidence interval (CI) were calculated using fixed- or random-effects model based on between-study heterogeneity. Results: A total of 44 eligible studies with 5,964 patients were included. The pooled results indicated a positive association of HIF-1α overexpression with poor overall survival (OS) (HR=1.990, 95% CI: 1.615-2.453, p<0.001) and disease-free survival (DFS) (HR=1.90, 95% CI: 1.084-3.329, p=0.043). Meta-analysis results showed that HIF-1α level expression was significantly associated with positive lymph node metastasis (OR=1.869, 95% CI: 1.488-2.248, p<0.001), distance metastasis (OR=2.604, 95% CI: 1.500-4.519, p<0.001), tumor stage (OR=1.801, 95% CI: 1.437-2.257, p<0.001) and tumor size (OR=1.392. 95% CI: 1.068-1.815, p=0.014). Conclusion: This meta-data suggest that HIF-1α expression might serve as an independent prognostic marker and a promising therapeutic target in patients with digestive system malignancies.

Safety and Efficacy of Combined Intramuscular/Intranasal RAZI-COV PARS Vaccine Candidate Against SARS-CoV-2: A Preclinical Study in Several Animal Models.

Several vaccine candidates for COVID-19 have been developed, and few vaccines received emergency approval with an acceptable level of efficacy and safety. We herein report the development of the first recombinant protein-based vaccine in Iran based on the recombinant SARS-CoV-2 spike protein in its monomeric (encompassing amino acid 1-674 for S1 and 685-1211 for S2 subunits) and trimer form (S-Trimer) formulated in the oil-in-water adjuvant system RAS-01 (Razi Adjuvant System-01). The safety and immunity of the candidate vaccine, referred to as RAZI-COV PARS, were evaluated in Syrian hamster, BALB/c mice, Pirbright guinea pig, and New Zeeland white (NZW) rabbit. All vaccinated animals received two intramuscular (IM) and one intranasal (IN) candidate vaccine at 3-week intervals (days 0, 21, and 51). The challenge study was performed intranasally with 5×106 pfu of SARS-CoV-2 35 days post-vaccination. None of the vaccinated mice, hamsters, guinea pigs, or rabbits showed any changes in general clinical observations; body weight and food intake, clinical indicators, hematology examination, blood chemistry, and pathological examination of vital organs. Safety of vaccine after the administration of single and repeated dose was also established. Three different doses of candidate vaccine stimulated remarkable titers of neutralizing antibodies, S1, Receptor-Binding Domain (RBD), and N-terminal domain (NTD) specific IgG antibodies as well as IgA antibodies compared to placebo and control groups (P<0.01). Middle and high doses of RAZI-COV PARS vaccine significantly induced a robust and quick immune response from the third-week post-immunization. Histopathological studies on vaccinated hamsters showed that the challenge with SARS-CoV-2 did not induce any modifications in the lungs. The protection of the hamster was documented by the absence of lung pathology, the decreased virus load in the lung, rapid clearance of the virus from the lung, and strong humoral and cellular immune response. These findings confirm the immunogenicity and efficacy of the RAZI-COV PARS vaccine. Of the three tested vaccine regimens, the middle dose of the vaccine showed the best protective immune parameters. This vaccine with heterologous prime-boost vaccination method can be a good candidate to control the viral infection and its spread by stimulating central and mucosal immunity.

The Brassica Napus Extract (BNE)-Loaded PLGA Nanoparticles as an Early Necroptosis and Late Apoptosis Inducer in Human MCF-7 Breast Cancer Cells.

Loading of the Brassica napus extract (BNE) on PLGA nanoparticle (BNE-PNP) and study its necroptotic activity in human MCF7-breast cancer cells. Double emulsion solvent evaporation methods were used for synthesis of BNE-PNP and DLS, SEM, and surface Zeta-potential analysis were applied for defining the physicochemical properties of BNE-PNP. The cytotoxic impact of BNE-PNP nanoparticles was analyzed by MTT assay and expression of apoptotic (P53 and Cas-3) and necrotic (TNF-α) gene markers were measured by qPCR to evaluate the BNE-PNP-induced cell death type. The stable (-36.07 mV) BNE-PNP were synthesized at 71.07 nm dimension. They significantly decrease the count of metabolically active MCF7 cells (IC50: 170.94 µg/ml after 48 h). The BNE-PNP induced an early programmed necrotic (necroptosis) and late apoptotic death on the MCF7 cancer cells by up-regulating all the P53/TNF-α and Cas-3 gene expression, respectively. The BNE-PNP dose-dependently induced an early cell-selective necroptotic death. Since the necroptotic death is known as a biocompatible cellular death induction, the BNE-PNP have the potential to be used as a safe efficient anticancer compound.

Biosynthesis and characterisation of solid lipid nanoparticles and investigation of toxicity against breast cancer cell line.

Solid lipid nanoparticles (SLNs) comprise non-toxic surface-active lipidic agents combined with appropriate ratios of drugs or essential oils. The goal of this research was to investigate the effects of the SLN synthesised using essential oils of Foeniculum vulgare on the MCF-7 breast cancer cell line. SLNs were prepared by homogenisation and ultrasound techniques and characterised by dynamic light scattering (DLS), zeta potential assessment, and transmission electron microscopy (TEM). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT assay), flow-cytometry, and Acridine-Orange assay were employed for assessing the biological activities of the SLNs. The average particle size was 55.43 nm and the net surface charge was -29.54 ± 11.67 mV. TEM showed that the mean particle size was 33.55 nm and the synthesised SLNs had a uniform round morphology. The MTT assay showed that the prepared SLNs had high toxicity against MCF-7 cells and low toxicity against normal HUVECs cells. Flow-cytometry revealed a noteworthy rise in the subG1 peak of the cell cycle in the cancer cells treated with SLNs compared to the controls, indicating apoptosis in cancer cells. The results also showed discolouration in SLNs-treated cells, which further confirmed the induction of apoptosis and the toxicity of the SLNs against MCF-7 cells.

The Clinical Value of Serum Adropin Level in Early Detection of Diabetic Nephropathy.

BACKGROUND/AIMS: Adropin is a metabolic hormone secreted by the liver, brain, and many peripheral tissues and is involved in energy homeostasis and insulin sensitivity. Some reports have indicated a significant decrease in serum adropin levels in type 2 diabetic patients. However, the significance of a decline in adropin level in early detection of diabetic nephropathy (DN) remains to be clarified. The purpose of this study was to evaluate the serum levels of adropin in patients with type 2 diabetes with and without nephropathy. METHODS: A total of 135 unrelated subjects (including 45 diabetic patients with nephropathy, 45 without nephropathy, and 45 healthy controls) were enrolled in this study. Fasting venous blood samples were collected from all patients. Serum adropin levels of all cases were analyzed by an enzyme-linked immunosorbent assay method. The correlations of serum adropin levels with anthropometric and biochemistry variables were determined. Logistic regression was performed to assess the association of adropin with odds of nephropathy. A receiver operating characteristic (ROC) curve was obtained to explore the optimum serum adropin concentration in distinguishing diabetic patients with and without nephropathy. RESULTS: Diabetic patients with nephropathy showed lower serum adropin levels than those in patients without nephropathy and healthy controls (p < 0.001). Pearson correlation analysis indicated that serum adropin was negatively correlated with BMI, FBS, HbA1c, blood urea, creatinine, LDL, and ACR and positively correlated with HDL and albumin. Logistic regression analysis showed that serum adropin was correlated with decreased risk of developing diabetic nephropathy. Moreover, in ROC analysis, at cutoff value 3.20 (mg/dL) with an AUC = 0.830, adropin had 80% sensitivity and 60% specificity for distinguishing the diabetic nephropathy. CONCLUSIONS: This study demonstrates that decreased level of adropin is associated with renal dysfunction in patients with type 2 diabetes mellitus. Serum adropin concentrations may be used as a biomarker for early detection of diabetic nephropathy.

MOF-Mediated Synthesis of CuO/CeO2 Composite Nanoparticles: Characterization and Estimation of the Cellular Toxicity against Breast Cancer Cell Line (MCF-7).

A copper oxide/cerium oxide nanocomposite (CuO/CeO2, NC) was synthesized via a novel method using a metal-organic framework as a precursor. This nanomaterial was characterized by Fourier transform infrared spectroscopy (FTIR), powder X-ray diffraction (PXRD), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), dynamic light scattering size analysis (DLS), and zeta potential. The PXRD showed the successful synthesis of the CuO/CeO2 NC, in which the 2theta values of 35.55° (d = 2.52 Å, 100%) and 38.73° (d = 2.32 Å, 96%) revealed the existence of copper (II) oxide. FTIR analysis showed the CeO2, hydroxyl groups, absorbed water, and some residual peaks. The solid phase analysis by FESEM and TEM images showed mean particle sizes of 49.18 ± 24.50 nm and 30.58 ± 26.40 nm, respectively, which were comparable with crystallite size (38.4 nm) obtained from PXRD, but it appears the CuO/CeO2 NC was not evenly distributed and in some areas, showed it was highly agglomerated. The hydrodynamic size (750.5 nm) also showed the agglomeration of the CuO/CeO2 NCs in the solution, which had a negatively charged surface. The CuO/CeO2 NCs showed anti-proliferative activity against human breast cancer cell line (MCF-7) in a dose- and time-dependence way, while affecting normal cells less significantly.

Application of Response Surface Methodology for Optimizing the Therapeutic Activity of ZnO Nanoparticles Biosynthesized from Aspergillus niger.

In this study, the biosynthesis of zinc oxide nanoparticles using Aspergillus niger (A/ZnO-NPs) is described. These particles have been characterized by UV-Vis spectrum analysis, X-ray powder diffraction, field emission scanning electron microscopy, and transmission electron microscopy. To use this biosynthesized nanoparticle as an antiproliferative and antimicrobial agent, the IC50 value against the breast cancer cell line and inhibition zone against Escherichia coli were used to optimize the effect of two processing factors including dose of filtrate fungi cell and temperature. The biosynthesized A/ZnO-NPs had an absorbance band at 320 nm and spherical shapes. The mean particles size was 35 nm. RSM (response surface methodology) was utilized to investigate the outcome responses. The Model F-value of 12.21 and 7.29 implies that the model was significant for both responses. The contour plot against inhibition zone for temperature and dose showed that if the dose increases from 3.8 to 17.2 µg/mL, the inhibition zone increases up to 35 mm. As an alternative to chemical and/or physical methods, biosynthesizing zinc oxide NPs through fungi extracts can serve as a more facile and eco-friendly strategy. Additionally, for optimization of the processes, the outcome responses in the biomedical available test can be used in the synthesis of ZnO-NPs that are utilized for large-scale production in various medical applications.

Two Dimensional Anion Exchange-Size Exclusion Chromatography Combined with Mathematical Modeling for Downstream Processing of Foot and Mouth Disease Vaccine.

The production of high-quality purified virus particles in high quantities for vaccine preparation requires a scalable purification procedure in the downstream step. A purification scheme based on combined strong anion-exchange and size exclusion chromatography (2D-AEC-SEC) was developed for the production of non-structural protein-free foot and mouth disease vaccine, and the whole procedure was accomplished with 77.9% virus yield. Additionally, a mathematical modeling and a simulation approach based on a plate model of chromatography were developed and matched with the experimental chromatography data to improve prediction of retention behavior and save time in the development of the downstream scale-up method. The purified pooled virus fraction obtained from the final polishing step had a purity higher than 85% based on analytical size exclusion analysis. Moreover, more than 90.1% of residual DNA (rDNA) was removed from the purified vaccine. The analysis of purified virus particles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), dynamic light scattering (DLS), high performance size exclusion chromatography (HP-SEC), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and transmission electron microscopy (TEM) provided clear evidence of purity and demonstrated that the final product is structurally spherical, intact particles qualified for formulation as a vaccine product.