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.

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.

Comparative Study on the Biological Effects of Sodium Citrate-Based and Apigenin-Based Synthesized Silver Nanoparticles.

The apigenin is a bioactive flavonoid mostly found in fruits and vegetables that possess various biological activities. The current study was performed to compare the biological potentials of sodium citrate-based (SC-SNPs) and apigenin-based (AP-SNPs) synthesized silver nanoparticles under the in vitro and in vivo conditions. The synthesized silver nanoparticles were physically and chemically characterized. The anticancer, pro-apoptotic, and their anti-bacterial activities were determined. Further, the mice trial was conducted to determine the possible toxic effects of the synthesized silver nanoparticles. The result of particle size analysis revealed the nanometer sizes of the SC-SNPs and AP-SNPs were about 95.5 and 93.94 nm, respectively. Both nanoparticles indicated pseudo-spherical shape, homogenous dispersion with an appropriate good degree of stability. However, the anticancer potential, pro-apoptotic effects and antibacterial activity of AP-SNPs were higher than that of SC-SNPs. Moreover, the mice trial indicated that AP-SNPs improved the liver function through modulation of liver enzymes, lipid peroxidation, and increase in the expression of antioxidant enzymes (SOD and GPx) as compared to the mice received AP-SNPs during 30 day experiment. Consequently the synthesis of silver nanoparticles using apigenin as reducing bioactive compound may result in production of silver nanoparticles with enhanced anticancer, antibacterial and antioxidant activities.

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.

Magnetic dispersive solid-phase extraction based on graphene oxide/Fe3 O4 @polythionine nanocomposite followed by atomic absorption spectrometry for zinc monitoring in water, flour, celery and egg.

BACKGROUND: Magnetic graphene oxide nanocomposite has been proposed as a promising and sustainable sorbent for the extraction and separation of target analytes from food matrices. Sample preparation based on nanocomposite presents several advantages, such as desired efficiency, reasonable selectivity and high surface-area-to-volume ratio. RESULTS: A new graphene oxide/Fe3 O4 @polythionine (GO/Fe3 O4 @PTh) nanocomposite sorbent was introduced for magnetic dispersive solid-phase extraction and flame atomic absorption spectrometric detection of zinc(II) in water, flour, celery and egg. To fabricate the sorbent, an oxidative polymerization of thionine on the surface of magnetic GO was applied, while polythionine was simply employed as a surface modifier to improve extraction yield. The properties of the sorbent were characterized by transmission electron microscopy, scanning electron microscopy, X-ray diffraction, energy-dispersive X-ray analysis, vibrating sample magnetometry and Fourier transform-infrared spectroscopy. The calibration curve showed linearity in the range of 0.5-30 ng mL-1 . Limits of detection (S/N = 3) and quantification (S/N = 10) were 0.08 and 0.5 ng mL-1 , respectively. CONCLUSION: The method was applied for trace-level determination of Zn(II) in water and food samples, and its validation was investigated by recovery experiments and analyzing certified reference material. © 2018 Society of Chemical Industry.

Sol-gel-based SPME fiber as a reliable sampling technique for studying biogenic volatile organic compounds released from Clostridium tetani.

A novel and efficient headspace solid-phase microextraction (HS-SPME) method, followed by gas chromatography mass spectrometry (GC-MS), was developed to study volatile organic compounds (VOCs) emerging from microorganisms. Two homemade SPME fibers, a semi-polar poly (dimethylsiloxane) (PDMS) fiber, and a polar polyethylene glycol (PEG) fiber, along with two commercial fibers (PDMS and PDMS/DVB) were used to collect VOCs emerging from Clostridium tetani which was cultured in different media. The adsorbed VOCs were desorbed and identified, in vitro, using GC-MS. The adsorption efficiency was improved by optimizing the time duration of adsorption and desorption. About 50 components were identified by the proposed method. The main detected compounds appeared to be sulfur containing compounds such as butanethioic acid S-methyl ester, dimethyl trisulfide, and dimethyl tetrasulfide. These volatile sulfur containing compounds are derived from amino acids containing the sulfur element, which probably coexist in the mentioned bacterium or are added to the culture media. The developed HS-SPME-GC-MS method allowed the determination of the chemical fingerprint of Clostridium tetani volatile constituents, and thus provides a new, simple, and reliable tool for studying the growth of microorganisms. Graphical abstract Investigation of biogenic VOCs released from Clostridium tetani using SPME-GC-MS.

Microwave-assisted extraction and high-throughput monolithic-polymer-based micro-solid-phase extraction of organophosphorus, triazole, and organochlorine residues in apple.

A high-throughput micro-solid-phase extraction device based on a 96-well plate was constructed and applied to the determination of pesticide residues in various apple samples. Butyl methacrylate and ethylene glycol dimethacrylate were copolymerized as a monolithic polymer and placed in the cylindrically shaped stainless-steel meshes of 96-micro-solid-phase extraction device and used as an extracting unit. Before the micro-solid-phase extraction, microwave-assisted extraction was employed to facilitate the transfer of the pesticide residues from the apple matrix to liquid media. Then, 1 mL of the aquatic samples was transferred into the 96-well plate and the 96-micro-solid-phase extraction device was applied for the extraction of the selected pesticides. Influential parameters, such as sorbent-to-sorbent reproducibility, microwave-assisted extraction time, ionic strength and micro-solid-phase extraction time, were optimized. The limits of quantitation were below 120 µg/kg, which are lower than the maximum residue limits. The developed method was successfully implemented for the extraction and determination of the selected pesticides from 20 different apple samples gathered from local markets. Phosalone was identified and quantified at the concentration level of 147 (±16.4) µg/kg in one of the samples.

Resorcinol-formaldehyde xerogel as a micro-solid-phase extraction sorbent for the determination of herbicides in aquatic environmental samples.

In this study, organic aerogels were synthesized by the sol-gel polycondensation of mixed cresol with formaldehyde in a slightly basic aqueous solution. Carbon aerogels and xerogels are generated by pyrolysis of organic aerogels. The novel sol-gel-based micro-solid-phase extraction sorbent, resorcinol-formaldehyde xerogel, was employed for preconcentration of some selected herbicides. Three herbicides of the aryloxyphenoxypropionate group, clodinafop-propargyl, haloxyfop-etotyl, and fenoxaprop-P-ethyl, were extracted from aqueous samples by micro-solid-phase extraction and subsequently determined by gas chromatography with mass spectrometry. The effect of different parameters influencing the extraction efficiency of these herbicides including sample flow rate, sample volume, and extraction time were investigated and optimized. Under optimum conditions, linear calibration curves in the range of 0.10-500 ng/L with R(2) > 0.99 were obtained. The relative standard deviation at 50 µg/L concentration level was lower than 10% (n = 5) and detection limits were between 0.05 and 0.20 µg/L. The proposed method was successfully applied to the sampling and extraction of herbicides from Zayanderood and paddy water samples.

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.

Amyloid fibrillation in native and chemically-modified forms of carbonic anhydrase II: role of surface hydrophobicity.

Chemical modification or mutation of proteins may bring about significant changes in the net charge or surface hydrophobicity of a protein structure. Such events may be of major physiological significance and may provide important insights into the genetics of amyloid diseases. In the present study, fibrillation potential of native and chemically-modified forms of bovine carbonic anhydrase II (BCA II) were investigated. Initially, various denaturing conditions including low pH and high temperatures were tested to induce fibrillation. At a low pH of around 2.4, where the protein is totally dissociated, the apo form was found to take up a pre-molten globular (PMG) conformation with the capacity for fibril formation. Upon increasing the pH to around 3.6, a molten globular (MG) form became abundant, forming amorphous aggregates. Charge neutralization and enhancement of hydrophobicity by methylation, acetylation and propionylation of lysine residues appeared very effective in promoting fibrillation of both the apo and holo forms under native conditions, the rates and extents of which were directly proportional to surface hydrophobicity, and influenced by salt concentration and temperature. These modified structures underwent more pronounced fibrillation under native conditions, than the PMG intermediate form, observed under denaturing conditions. The nature of the fibrillation products obtained from intermediate and modified structures were characterized and compared and their possible cytotoxicity determined. Results are discussed in terms of the importance of surface net charge and hydrophobicity in controlling protein aggregation. A discussion on the physiological significance of the observations is also presented.

Determination of free formaldehyde in vaccines and biological samples using solid-phase microextraction coupled to GC-MS.

A headspace solid-phase microextraction method coupled to GC-MS was successfully developed for the trace determination of formaldehyde in veterinary bacterial and human vaccines, and diphtheria-tetanus antigen. The formaldehyde was derivatized by means of the Hantzsch reaction prior to extraction and subsequent determination. Three different types of solid-phase microextraction fibers, polar, and nonpolar poly(dimethylsiloxane) and polyethylene glycol were prepared by using a sol-gel technique. The effects of different parameters such as type of fiber coating, extraction time and temperature, desorption conditions, agitation rate, and salt effect were investigated. Under the optimized conditions, the detection limit of the method was 979 ng/L using the selected ion-monitoring mode. The interday and intraday precisions of the developed method under the optimized conditions were below 13%, and the method shows linearity in the range of 1.75-800 µg/L with a correlation coefficient of 0.9963. The optimized method was applied to the determination of formaldehyde from some biological products. The results were satisfactory compared to the standard method.

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.

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.

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).

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.