Targeting Leishmaniasis with Nitrovinyl Derivatives: Their Synthesis, In Vitro Assessment, and Computational Exploration.

INTRODUCTION: Leishmaniasis is an affliction caused by the protozoan parasites of the Leishmania genus. This disease impacts a substantial global populace, exceeding one million individuals, leading to disability-adjusted life years and fatalities, particularly within tropical regions. At present, the existing drug therapies have not attained a degree of efficacy that can be unequivocally classified as genuinely triumphant. In this context, the conception of novel compounds possessing the capacity to impede the parasite's life cycle at various stages holds considerable significance. METHODS: In this research endeavor, an exploration was undertaken involving the design and synthesis of nineteen derivatives incorporating the nitrovinyl pharmacophore. The subsequent evaluation of their impacts on L. major was conducted through a combination of in vitro (amastigote and promastigote inhibition) and in silico (molecular docking) investigations. RESULTS: All of the compounds were synthesized and purified with good yields. In the amastigote inhibition assay, compounds 10, 15, and 18 showed better inhibitory effects than the standard drug meglumine antimonate (MA). Regarding the synergistic impact of synthesized compounds and MA together, all outcomes were significantly better than those of monotherapy of each in amastigote and macrophage forms. In the promastigote assay, compounds 2, 8, 12, 15, 16, 17, and 19 demonstrated superior inhibitory effects compared to MA. Moreover, compounds 4, 12, and 15 showed the best synergies with MA in inhibiting amastigotes. According to docking scores, 1XTP (a SAM-dependent methyltransferase) and 4G5D (Prostaglandin F synthase) receptors were found to be the most probable targets in their mechanism of action. CONCLUSION: In vitro evaluations and computational analyses strongly suggest that these compounds could be effective against both L. major amastigotes and promastigotes. Additionally, they exhibited notable synergistic interactions with MA against both living forms of the parasite.

Medium optimization to improve growth and iron uptake by Bacillus tequilensis ASFS1 using fractional factorial designs.

Many notable applications have been described for magnetic nanoparticles in delivery of diverse drugs and bioactive compounds into cells, magnetofection for the treatment of cancer, photodynamic therapy, photothermal therapy, and magnetic particle imaging (MPI). In response to the growing demand for magnetic nanoparticles for drug delivery or biomedical imaging applications, more effective and eco-friendly methodologies are required for large-scale biosynthesis of this nanoparticles. The major challenge in the large-scale biomedical application of magnetic nanoparticles lies in its low efficiency and optimization of nanoparticle production can address this issue. In the current study, a prediction model is suggested by the fractional factorial designs. The present study aims to optimize culture media components for improved growth and iron uptake of this strain. The result of optimization for iron uptake by the strain ASFS1 is to increase the production of magnetic nanoparticles by this strain for biomedical applications in the future. In the present study, design of experiment method was used to probe the effects of some key medium components (yeast extract, tryptone, FeSO4, Na2-EDTA, and FeCl3) on Fe content in biomass and dried biomass of strain ASFS1. A 25-1 fractional factorial design showed that Na2-EDTA, FeCl3, yeast extract-tryptone interaction, and FeSO4-Na2-EDTA interaction were the most parameters on Fe content in biomass within the experimented levels (p < 0.05), while yeast extract, FeCl3, and yeast extract-tryptone interaction were the most significant factors within the experimented levels (p < 0.05) to effect on dried biomass of strain ASFS1. The optimum culture media components for the magnetic nanoparticles production by strain ASFS1 was reported to be 7.95 g L-1 of yeast extract, 5 g L-1 of tryptone, 75 µg mL-1 of FeSO4, 192.3 µg mL-1 of Na2-EDTA and 150 µg mL-1 of FeCl3 which was theoretically able to produce Fe content in biomass (158 µg mL-1) and dried biomass (2.59 mg mL-1) based on the obtained for medium optimization. Using these culture media components an experimental maximum Fe content in biomass (139 ± 13 µg mL-1) and dried biomass (2.2 ± 0.2 mg mL-1) was obtained, confirming the efficiency of the used method.

Purification, Characterization, and Assessment of Anticancer Activity of Iron Oxide Nanoparticles Biosynthesized by Novel Thermophilic Bacillus tequilensis ASFS1 .

Magnetic nanoparticles (MNPs), particularly iron oxide nanoparticles (IONPs), are a fascinating group of nanoparticles that have been considerably investigated for biomedical applications because of their superparamagnetic properties, biodegradable nature, and biocompatibility. A novel Gram-positive moderately thermophilic bacterial strain, namely Bacillus tequilensis ASFS.1, was isolated and identified. This strain is capable of producing superparamagnetic Fe3O4 nanoparticles and exhibiting magnetotaxis behavior. This strain swimming behavior was investigated under static and dynamic environments, where it behaved very much similar to the magnetotaxis in magnetotactic bacteria. This study is the first report of a bacterium from the Bacillaceae family that has the potential to intracellular biosynthesis of IONPs. MNPs were separated by a magnetic and reproducible method which was designed for the first time for this study. In addition, UV-visible spectrophotometer, Fourier-transform infrared spectroscopy, vibrating sample magnetometer, field emission scanning electron microscopy (FESEM), X-ray diffraction, and thermal gravimetric analysis were utilized to characterize the bio-fabricated magnetite nanoparticles. Analysis of the particle size distribution pattern of the biogenic MNPs by FESEM imaging revealed the size range of 10-100 nm with the size range of 10-40 nm MNPs being the most frequent particles. VSM analysis demonstrated that biogenic MNPs displayed superparamagnetic properties with a high saturation magnetization value of 184 emu/g. After 24 h treatment of 3T3, U87, A549, MCF-7, and HT-29 cell lines with the biogenic MNPs, IC50 values were measured to be 339, 641, 582, 149, and 184 µg mL-1, respectively. This study presents the novel strain ASFS.1 capable of magnetotaxis by the aid of its magnetite nanoparticles and paving information on isolation, characterization, and in vitro cytotoxicity of its MNPs. The MNPs showed promising potential for biomedical applications, obviously subject to additional studies.

Neuroprotective Effects of [Formula: see text]-Terpinene in Rats with Acute Cerebral Ischemia: Modulation of Inflammation, Apoptosis, and Oxidation.

The study aimed to assess 𝛾-Terpinene's (𝛾-TER) neuroprotective potential in acute cerebral ischemia, characterized by reduced cerebral blood flow in rats. Middle cerebral artery occlusion (MCAO), a standard method for inducing cerebral ischemia, was employed in male Wistar rats. 𝛾-TER at varying doses (5, 10, and 15 mg/kg) were intraperitoneally administered during reperfusion onset. Neurological outcomes, cerebral infarct size, edema, and enzymatic activities (SOD, GPx, and catalase) in the brain were evaluated using diverse techniques. The study examined gene expression and pathways associated with neuroinflammation and apoptosis using Cytoscape software, identifying the top 10 genes involved. Pro-inflammatory and pro-apoptotic factors were assessed through real-time PCR and ELISA, while apoptotic cell rates were measured using the TUNEL and Flow cytometry assay. Immunohistochemistry assessed apoptosis-related proteins like Bax and bcl-2 in the ischemic area. 𝛾-TER, particularly at doses of 10 and 15 mg/kg, significantly reduced neurological deficits and cerebral infarction size. The 15 mg/kg dose mitigated TNF-α, IL-1ß, Bax, and caspase-3 gene and protein levels in the cortex, hippocampus, and striatum compared to controls. Furthermore, Bcl-2 levels increased in these regions. 𝛾-TER show cased neuroprotective effects by suppressing inflammation, apoptosis, and oxidation. In conclusion, 𝛾-TER, possessing natural anti-inflammatory and anti-apoptotic properties, shields the brain against ischemic damage by reducing infarction, edema, oxidative stress, and inflammation. It modulates the expression of crucial genes and proteins associated with apoptosis in diverse brain regions. These findings position 𝛾-TER as a potential therapeutic agent for ischemic stroke.

Bedsore Healing Using Selenium-Enriched Lactobacillus brevis LSe: A Randomized, Double-Blind, Controlled Clinical Trial.

Bedsores impose an important challenge to the healthcare system. Se-baring probiotics are considered effective agents in wound healing and inflammation reduction via several pathways. The present study focused on the administration of a Se-enriched probiotic, originally obtained from a traditional dairy product for bedsore healing. Daily doses of the probiotic were administered to 20 ICU patients for 14 days and the wound healing criteria were compared with those of the same group of ICU patients as control, both groups suffering from stages I and II bedsore (a randomized, double-blind, controlled clinical trial). The administered Se-enriched probiotic decreased the bedsore healing period significantly (on average by 2.4 days, P-value: 0.039), as well as bedsore size (on average by 7 mm2, nonsignificant) and bedsore grade (10%, nonsignificant) in the treatment group more efficiently than the control group. Some key laboratory parameters associated with inflammation were also improved in patients receiving the Se-supplemented probiotic. The limitations of this study include the low number of patients meeting inclusion criteria within the timeframe of the study, and the impossibility of following up patients after discharge from the ICU. In summary, this study revealed the effectiveness of the Se-enriched probiotic in bedsore improvement, suggesting consideration of the enriched probiotic as an auxiliary agent in bedsore management.

A qualitative study on barriers and strategies to hospital preparedness against chemical, biological, radiological, and nuclear incidents.

BACKGROUND: The effects of chemical, biological, radiological, and nuclear (CBRN) incidents on human society can be irreparable. Preparing the health system for these incidents is essential. This study aims to identify obstacles to hospital preparedness against CBRN incidents and provide solutions. MATERIALS AND METHODS: This qualitative study was conducted using semi-structured interview method in 2022. The research community included experts in the fields of CBRN, and 17 persons were included in the study through purposive sampling. The interviews were collected by interview guide and recorded face-to-face and online and were analyzed using thematic content analysis method in MS Word 2016. RESULTS: The interviews' analysis was classified into two main categories and 34 sub-categories. Some of the most important obstacles to the hospital preparedness against CBRN incidents were the lack of proper crisis management, the lack of specialist staff, the stress of employees, the lack of turnover and sufficient rest for employees, legal gaps, and so on. The main strategies were determining the type and extent of the risk factor, strong crisis management, the lack of parallelism, continuous monitoring, having a protocol and road map, appropriate training programs, having skilled personnel, rapid response of personnel, positive attitude of the staff, and the favorable condition of the building. CONCLUSION: The appointment of an expert in the field of CBRN and having a specialized unit, the existence of specialized and trained staff along with access to the required facilities, clear instructions, and intra-departmental and inter-departmental cooperation affect the readiness of hospitals against CBRN incidents.

Mutuality of epigenetic and nanoparticles: two sides of a coin.

Nowadays nanoparticles (NPs) due to their multidimensional applications in enormous different fields, has become an exciting research topic. In particular, they could attract a noticeable interest as drug deliver with increased bioavailability, therapeutic efficacy and drug specificity. Epigenetic can be considered as a complex network of molecular mechanism which are engaged in gene expression and have a vital role in regulation of environmental effects on ethology of different disorders like neurological disorders, cancers and cardiovascular diseases. For many of them epigenetic therapy was proposed although its application accompanied with limitations, due to drug toxicity. In this review we evaluate two aspects to epigenetic in the field of NPs: firstly, the role of epigenetic in regulation of nanotoxicity and secondly application of NPs as potential carriers for epidrugs.

Antileishmanial effects and drugability characteristics of a heterocyclic copper complex: An in silico, in vitro and molecular study.

Leishmaniasis caused by the protozoan Leishmania presents a severe illness, principally in tropical and subtropical areas. Antileishmanial metal complexes, like Glucantime®ï¸ with proven activity, are routinely studied to probe their potency. We investigated the effects of a Cu (II) homoleptic complex coordinated by two dimethyl-bipyridine ligands against Leishmania major stages in silico and in vitro. The affinity of this heterocyclic Cu (II) complex (CuDMBP) towards a parasitic metacaspase was studied by molecular docking. Key pharmacokinetic and pharmacodynamic properties of the complex were predicted using three web-based tools. CuDMBP was tested for in vitro antileishmanial activities using MTT assay, model murine macrophages, flow cytometry, and quantitative real-time polymerase chain reaction (qPCR). Molecular docking confirmed the tendency between the target macromolecule and the complex. ADMET evaluations highlighted CuDMBP's key pharmacological features, including P-glycoprotein-associated GI absorption and lack of trans-BBB permeability. MTT showed significant inhibitory effects against promastigotes. CuDMBP significantly increased the level of cellular IL-12 expression (p < 0.05), while the upregulation observed in the expression of iNOS was considered not significant (p > 0.05). It decreased the expression of IL-10 significantly (p < 0.05). Findings demonstrated that CuDMBP deserves to be introduced as a leishmanicidal candidate provided further studies are carried out.

An insight into biofabrication of selenium nanostructures and their biomedical application.

Evidence shows that nanoparticles exert lower toxicity, improved targeting, and enhanced bioactivity, and provide versatile means to control the release profile of the encapsulated moiety. Among different NPs, inorganic nanoparticles (Ag, Au, Ce, Fe, Se, Te, Zn, etc.) possess a considerable place owing to their unique bioactivities in nanoforms. Selenium, an essential trace element, played a vital role in the growth and development of living organisms. It has attracted great interest as a therapeutic factor without significant adverse effects in medicine at recommended dose. Selenium nanoparticles can be fabricated by physical, biological, and chemical approaches. The biosynthesis of nanoparticles is shown an advance compared to other procedures, because it is environmentally friendly, relatively reproducible, easily accessible, biodegradable, and often results in more stable materials. The effect of size, shape, and synthesis methods on their applications in biological systems investigated by several studies. This review focused on the procedures for the synthesis of selenium nanoparticles, in particular the biogenesis of selenium nanoparticles and their biomedical characteristics, such as antibacterial, antiviral, antifungal, and antiparasitic properties. Eventually, a comprehensive future perspective of selenium nanoparticles was also presented.

Innovative Diagnostic Peptide-Based Technologies for Cancer Diagnosis: Focus on EGFR-Targeting Peptides.

Active targeting using biological ligands has emerged as a novel strategy for the targeted delivery of diagnostic agents to tumor cells. Conjugating functional targeting moieties with diagnostic probes can increase their accumulation in tumor cells and tissues, enhancing signal detection and, thus, the sensitivity of diagnosis. Due to their small size, ease of chemical synthesis and site-specific modification, high tissue penetration, low immunogenicity, rapid blood clearance, low cost, and biosafety, peptides offer several advantages over antibodies and proteins in diagnostic applications. Epidermal growth factor receptor (EGFR) is one of the most promising cancer biomarkers for actively targeting diagnostic and therapeutic agents to tumor cells due to its active involvement and overexpression in various cancers. Several peptides for EGFR-targeting have been identified in the last decades, which have been obtained by multiple means including derivation from natural proteins, phage display screening, positional scanning synthetic combinatorial library, and in silico screening. Many studies have used these peptides as a targeting moiety for diagnosing different cancers in vitro, in vivo, and in clinical trials. This review summarizes the progress of EGFR-targeting peptide-based assays in the molecular diagnosis of cancer.

A Novel Hyperthermostable Recombinant Protein Nanocage

Background: Background: Ferritin has an important role in iron storage in the cells, and due to its nanocage structure and self-assembly properties, it has wide application prospects in nanobiotechnology. Methods: Methods: The maize (Zea mays) ferritin gene ZmFer1 was cloned and expressed in Escherichia coli BL21 (DE3) for the first time. Change in macromolecular structure of ZmFer1 ferritin due to heat treatment was investigated using native PAGE electrophoresis, dynamic light scattering (DLS), and transmission electron microscopy (TEM). Change in the secondary structures of the protein was evaluated using circular dichroism spectroscopy. Moreover, alteration in the conformation of the protein was evaluated using UV-absorption spectra and intrinsic fluorescence spectra. The melting temperature (Tm) of ZmFer1 was obtained using differential scanning calorimetry (DSC). Finally, the effect of heat on the function of ZmFer1 was assessed by iron loading ability. Results: Results: The purified ZmFer1 protein showed a homopolymer nanocage structure. The results of native PAGE electrophoresis, DLS, and TEM techniques showed that ZmFer1 protein nanocage is stable to heat treatment up to 90 °C, and some of the protein nanocages retain their macromolecular structures even at 100 °C in liquid aqueous solution. Based on the DSC results, ZmFer1 protein nanocage had a Tm of 81.9 °C. After treatment at 100 °C, stable ZmFer1 protein nanocages were able to store iron atoms. Conclusion: Conclusion: Recombinant ZmFer1 ferritin with a Tm > 80°C is a hyperthermostable protein nanocage. The results of this study are beneficial for the development of protein nanocages that are stable under extreme temperature conditions, as well as application of ZmFer1 in nanobiotechnology, biomaterials, and biomedical fields.

In vitro biological and in silico molecular docking and ADME studies of a substituted triazine-coordinated cadmium(II) ion: efficient cytotoxicity, apoptosis, genotoxicity, and nuclease-like activity plus binding affinity towards apoptosis-related proteins.

A cadmium(II) complex containing dppt ligand with the formula [CdCl2(dppt)2], where dppt is 5,6-diphenyl-3-(2-pyridyl)-1,2,4-triazine was synthesized, elucidated and submitted to in vitro cytotoxicity studies against human breast (MCF-7), glioblastoma (U-87), and lung (A549) cancer cell lines as well as mouse embryo normal cell line (NIH/3T3), in comparison with cisplatin employing MTT assay over 24 and 48 h. The complex exhibited the highest cytotoxic effect against MCF-7 cells among the other three cell lines with IC50 values of 8.7 ± 0.5 (24 h) and 1.2 ± 0.7 µM (48 h). Significantly, flow cytometric assessment of the complex-treated MCF-7 and U-87 cells demonstrated a dose-dependent induced apoptotic cell death. The cellular morphological changes were in concord with cytotoxicity and flow cytometric results. The results of comet assay showed that the complex is able to induce DNA damage in MCF-7 cells. These observations are of importance, as sustained damage to cellular DNA could lead to apoptotic cell death. The results of DNA-binding studies indicated that the complex fits into the DNA minor groove and interacts with DNA via a partial intercalation. Moreover, the complex was able to efficiently cleave pUC19 DNA through a hydrolytic mechanism. The binding affinity between the complex and apoptosis-relevant protein targets including APAF1, Bax, Bcl-2, Cas3, Cas7, and Cas9 was evaluated through molecular docking studies. In silico virtual studies revealed the complex's strong affinity towards apoptosis-related proteins; therefore the complex can act as a potential apoptosis inducer. Physicochemical, pharmacokinetics, lipophilicity, drug-likeness, and medicinal chemistry properties of the complex were also predicted through in silico absorption, distribution, metabolism and excretion studies.

In silico and in vitro antileishmanial effects of gamma-terpinene: Multifunctional modes of action.

INTRODUCTION: Leishmaniasis denotes a significant health challenge worldwide with no ultimate treatment. The current study investigated the biological effects of gamma-terpinene (GT) on Leishmania major in putative antileishmanial action, cytotoxicity, apoptosis induction, gene expression alteration, antioxidant activity, hemolysis, and ROS generation. METHODS: GT and meglumine antimoniate (MA) were probed alone and in combination (GT/MA) for their anti-leishmanial potentials using the MTT biochemical colorimetric assay and a model macrophage cell. In addition, their immunomodulatory properties were assessed by analyzing their effect on the transcription of cytokines related to Th1 and Th2 responses. GT and MA, alone and in combination, were also assessed for their potential to alter metacaspase gene expression in L. major promastigotes by real-time RT-PCR. The hemolytic potential of GT and MA-treated promastigotes were also measured by routine UV absorbance reading. Electrophoresis on agarose gel was employed to analyze genomic DNA fragmentation. RESULTS: GT demonstrated notable dose-dependent antileishmanial effects towards promastigotes and amastigotes of L. major. The IC50 values for GT against L. major promastigotes and amastigotes were 46.76 mM and 25.89 mM, respectively. GT was considerably safer towards murine macrophages than L. major amastigotes with an SI value of 3.17. Transcriptional expression of iNOS, JAK-1, Interleukin (IL-10), and TGF-ß was meaningfully decreased, while the levels of metacaspase mRNA were increased. Results also confirmed GT antioxidant activities. Also, increased levels of intracellular ROS were observed upon treatment of promastigotes with GT. The gel electrophoresis result indicated slight DNA fragmentation in the treated promastigotes by both drugs. A weak hemolytic effect was also observed for GT. CONCLUSION: The results demonstrated that GT showed potent activity against L. major stages. It seems that its mechanism of action involves representing an immunomodulatory role towards upregulation of iNOS and JAK-1, while downregulation of IL-10 and TGF- ß. Moreover, GT has an antioxidative potential and exerts its action through activating macrophages to kill the organism. Further in vivo and clinical studies are essential to explore its effect in future programs.

Nuclease-like metalloscissors: Biomimetic candidates for cancer and bacterial and viral infections therapy.

Despite the extensive and rapid discovery of modern drugs for treatment of cancer, microbial infections, and viral illnesses; these diseases are still among major global health concerns. To take inspiration from natural nucleases and also the therapeutic potential of metallopeptide antibiotics such as the bleomycin family, artificial metallonucleases with the ability of promoting DNA/RNA cleavage and eventually affecting cellular biological processes can be introduced as a new class of therapeutic candidates. Metal complexes can be considered as one of the main categories of artificial metalloscissors, which can prompt nucleic acid strand scission. Accordingly, biologists, inorganic chemists, and medicinal inorganic chemists worldwide have been designing, synthesizing and evaluating the biological properties of metal complexes as artificial metalloscissors. In this review, we try to highlight the recent studies conducted on the nuclease-like metalloscissors and their potential therapeutic applications. Under the light of the concurrent Covid-19 pandemic, the human need for new therapeutics was highlighted much more than ever before. The nuclease-like metalloscissors with the potential of RNA cleavage of invading viral pathogens hence deserve prime attention.

Immobilization of Clostridium perfringens type D in calcium alginate beads: toxin production mimics free cell culture.

Background and Objectives: Cell-immobilization is used to maintain microbial culture to produce metabolites in repeated-batch or continuous fermentations, thereby reducing the time and resources spent on delivering mass production of microbe. The technique also enables shortening of the detoxification phase and the amount of formaldehyde required due to low incidence of viable bacteria in the extract. Materials and Methods: A solution of sodium alginate containing Clostridium perfringens cells was dropped into stirring CaCl solution via a sterile syringe needle. Optimizations resulted in reasonably uniform beads containing C. perfringens. Beads were externally stabilized by poly L-lysine, followed by immersion in a solution of Na-alginate to coat them with a new layer of alginate forming an alginate-PLL-alginate cortex. Results: This study proved successful in immobilizing C. perfringens cells inside uniform alginate microspheres. Cell loading and cell propagation inside the beads were measured. The cell loaded beads were cultivable in liquid media producing 550 minimum lethal doses per milliliter (MLD/ml) in a 72 h. Conclusion: The research paved the way for further investigations to optimize and establish an efficient bacterial encapsulation method. Thus, it seems possible to produce toxins from beads engulfing C. perfringens on larger scales via repeated-batch or continuous fermentation processes.

Rapid microwave-assisted biosynthesis of platinum nanoparticles and evaluation of their antioxidant properties and cytotoxic effects against MCF-7 and A549 cell lines.

In this study, platinum nanoparticles (Pt NPs) were synthesized by a green method using an aqueous extract of Eucalyptus camaldulensis with assistance of microwave irradiation (850 W) and their physicochemical characteristics were studied by UV-visible spectroscopy, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) analyses. Antioxidant activities, hemocompatibility, and cytotoxic effects of the prepared Pt NPs were then evaluated. The attained results showed that the newly formed Pt NPs possess a size range between 7.4 and 11.2 nm. These spherical-shaped NPs were slightly aggregated and held various functional groups on their surface. The antioxidant activity of Pt nanostructures was comparable to that of butylated hydroxyl anisole at concentrations higher than 320 µg/mL. At the same concentration of 640 µg/mL, the scavenging activities were 3.36 ± 0.9% (hexachloroplatinic acid) and 52.13 ± 0.43% (Pt NPs). The results of hemolytic assay revealed satisfactory hemocompatibility of the Pt NPs even at the concentration as high as 4 mg/mL (hemolysis percent equal to 3.5 ± 1.3%). The cytotoxicity studies revealed that MCF-7, A549, and 3T3 cell lines treated with hexachloroplatinic acid and cisplatin for 24 h and 48 h showed a higher percentage of cell death compared with the Pt NPs. After 24 h, for A549, 3T3, and MCF-7 cells exposed to Pt NPs, the cell viability was measured to be 80 ± 3.2%, 96 ± 1%, and 89 ± 2.6%, respectively, at concentration of 640 µg/mL. Further investigations are required to elucidate the mechanisms behind the biological activities of as-synthesized Pt NPs.

A brief review on DNA vaccines in the era of COVID-19.

This article provides a brief overview of DNA vaccines. First, the basic DNA vaccine design strategies are described, then specific issues related to the industrial production of DNA vaccines are discussed, including the production and purification of DNA products such as plasmid DNA, minicircle DNA, minimalistic, immunologically defined gene expression (MIDGE) and Doggybone™. The use of adjuvants to enhance the immunogenicity of DNA vaccines is then discussed. In addition, different delivery routes and several physical and chemical methods to increase the efficacy of DNA delivery into cells are explained. Recent preclinical and clinical trials of DNA vaccines for COVID-19 are then summarized. Lastly, the advantages and obstacles of DNA vaccines are discussed.

Cytotoxicity of mancozeb on Sertoli-germ cell co-culture system: Role of MAPK signaling pathway.

Mancozeb (MZB) is a worldwide fungicide for the management of fungal diseases in agriculture and industrial contexts. Human exposure occurs by consuming contaminated plants, drinking water, and occupational exposure. There are reports on MZB's reprotoxicity such as testicular structure damage, sperm abnormalities, and decrease in sperm parameters (number, viability, and motility), but its molecular mechanism on apoptosis in testis remains limited. To investigate the molecular mechanisms involved in male reprotoxicity induced by MZB, we used primary cultures of mouse Sertoli-germ cells. Cells were exposed to MZB (1.5, 2.5, and 3.5 µM) for 3 h to evaluate viability by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, reactive oxygen species (ROS) generation, and oxidative stress parameters (lipid peroxidation). Cell death and mitogen-activated protein kinase (MAPK) signaling were measured in these cells using flow cytometry and western blotting. In addition, some groups were exposed to N-acetylcysteine (NAC, 5 mM) in the form of co-treatment with MZB. Mancozeb reduced viability and increased the level of intracellular ROS, p38 and c-Jun N-terminal kinases (JNK) MAPK proteins phosphorylation, and apoptotic cell death, which could be blocked by NAC as an inhibitor of oxidative stress. The present study indicated for the first time the toxic manifestations of MZB on the Sertoli-germ cell co-culture. Redox imbalance and p38 and JNK signaling pathway activation might play critical roles in MZB-induced apoptosis in the male reproductive system.

Optimization of immobilization of Pseudomonas cepacia lipase on multiwalled carbon nanotubes functionalized with glycyrrhizin and Tween 80.

In the present study, multiwalled carbon nanotubes (MWCNTs) were functionalized with glycyrrhizin and Tween 80 and applied for immobilization of Pseudomonas cepacia lipase (PcL). Characterization of f-MWCNTs was performed through Fourier-transform infrared spectroscopy, thermal gravimetric, field emission scanning electron microscopy, and energy-dispersive X-ray spectroscopy analysis. The optimum specific activity of immobilized PcL (studied by Plackett-Burman statistical design) occurred at 0.3 mg/mL of f-MWCNTs, 25 mM of phosphate buffer (pH 6.0), 15 min sonication time, 8 U/mL of enzyme concentration, and 24 h immobilization time at 4 °C in the absence of glutaraldehyde. In these conditions, the specific activity was 16.57 ± 0.71 U/mg, which was very close to the predicted amount (16.62 ± 0.64 U/mg). The results of thermal and pH stability showed that the stability of immobilized PcL was higher than that of the free PcL. The activity of immobilized PcL on f-MWCNTs held 93% after being incubated for 60 min at 70 °C. Moreover, the immobilized PcL on f-MWCNTs retained about 65% of its initial activity after 30 days of storage at 25 °C. In addition, about 50% of initial activity of immobilized PcL retained after 10 cycles of uses. Therefore, f-MWCNTs could be introduced as suitable support for enzymes immobilization.

In silico and in vitro inhibitory potential of an organometallic Cu (II) complex on Leishmania major stages.

Leishmaniosis results in a serious complication, principally in the tropical and subtropical areas. Metalcored complexes, like meglumine antimoniate (MA) have proven antileishmanial activity. Similarly, in this research, we investigated the effects of Cu (II) dimethoxy bipyridine (CuDMOBP) against Leishmania major stages in silico and in vitro. Molecular docking analysis was carried out on the complex and a protozoan metacaspase. The complex's antipromastigote and its cytotoxicity towards macrophages were assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method to calculate relative Inhibitory Concentration 50% (IC50), Cytotoxic Concentration 50% (CC50), and Selectivity Index (SI). Expression of TNF-α and IL-10 in intracellular amastigotes and induction of apoptosis was also investigated using quantitative real-time PCR. The complex interacted effectively with four amino acid residues including lysine (Lys171), histidine (His193), arginine (Arg44 and Arg243) of the targeted metacaspase. This indicates a potential affinity between the target macromolecule and the complex. MTT results showed significant in vitro inhibitory effects against promastigotes. Reduction in cellular expression of IL-10 and TNF-α was also significant, p<0.05 and p<0.005, respectively. CuDMOBP showed powerful in vitro anti-leishmanial activity and could be introduced as a new leishmanicidal candidate.