Thymus Gland: A Double Edge Sword for Coronaviruses.

The thymus is the main lymphoid organ that regulates the immune and endocrine systems by controlling thymic cell proliferation and differentiation. The gland is a primary lymphoid organ responsible for generating mature T cells into CD4+ or CD8+ single-positive (SP) T cells, contributing to cellular immunity. Regarding humoral immunity, the thymic plasma cells almost exclusively secrete IgG1 and IgG3, the two main complement-fixing effector IgG subclasses. Deformity in the thymus can lead to inflammatory diseases. Hassall's corpuscles' epithelial lining produces thymic stromal lymphopoietin, which induces differentiation of CDs thymocytes into regulatory T cells within the thymus medulla. Thymic B lymphocytes produce immunoglobulins and immunoregulating hormones, including thymosin. Modulation in T cell and naive T cells decrement due to thymus deformity induce alteration in the secretion of various inflammatory factors, resulting in multiple diseases. Influenza virus activates thymic CD4+ CD8+ thymocytes and a large amount of IFNγ. IFNs limit virus spread, enhance macrophages' phagocytosis, and promote the natural killer cell restriction activity against infected cells. Th2 lymphocytes-produced cytokine IL-4 can bind to antiviral INFγ, decreasing the cell susceptibility and downregulating viral receptors. COVID-19 epitopes (S, M, and N proteins) with ≥90% identity to the SARS-CoV sequence have been predicted. These epitopes trigger immunity for antibodies production. Boosting the immune system by improving thymus function can be a therapeutic strategy for preventing virus-related diseases. This review aims to summarize the endocrine-immunoregulatory functions of the thymus and the underlying mechanisms in the prevention of COVID-19.

Genome composition and genetic characterization of SARS-CoV-2.

SARS-CoV-2 is a type of Betacoronaviruses responsible for COVID-19 pandemic disease, with more than 1.745 million fatalities globally as of December-2020. Genetically, it is considered the second largest genome of all RNA viruses with a 5' cap and 3' poly-A tail. Phylogenetic analyses of coronaviruses reveal that SARS-CoV-2 is genetically closely related to the Bat-SARS Like-Corona virus (Bat-SL-Cov) with 96% whole-genome identity. SARS-CoV-2 genome consists of 15 ORFs coded into 29 proteins. At the 5' terminal of the genome, we have ORF1ab and ORF1a, which encode the 1ab and 1a polypeptides that are proteolytically cleaved into 16 different nonstructural proteins (NSPs). The 3' terminal of the genome represents four structural (spike, envelope, matrix, and nucleocapsid) and nine accessory (3a, 3b, 6, 7a, 7b, 8b, 9a, 9b, and orf10) proteins. As the number of COVID-19 patients increases dramatically worldwide, there is an urgent need to find a quick and sensitive diagnostic tool for controlling the outbreak of SARS-CoV-2 in the community. Today, molecular testing methods utilizing viral genetic material (e.g., PCR) represent the crucial diagnostic tool for the SARS-CoV-2 virus despite its low sensitivity in the early stage of viral infection. This review summarizes the genome composition and genetic characterization of the SARS-CoV-2.

Immunogenicity of Multiple Doses of pDNA Vaccines against SARS-CoV-2.

Since its identification in Wuhan, China, in December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), has resulted in 46 million cases and more than one million deaths worldwide, as of 30 October 2020. Limited data exist on the magnitude and durability of antibodies generated by natural infection with SARS-CoV-2 and whether they can provide long-lasting immunity from reinfection. Vaccination has proven the most effective measure for controlling and preventing pandemics and, thus, development of a vaccine against COVID-19 is a top priority. However, the doses required to induce effective, long-lasting antibody responses against SARS-CoV-2 remain undetermined. Here, we present the development of SARS-CoV-2 vaccine candidates encoding the viral spike (S) gene, generated using plasmid (p)DNA technology, and we demonstrate the eliciting of S-specific antibodies in mice after three and four doses. The magnitude of binding and neutralizing antibody responses with three doses of synthetic, codon-optimized, full-length S (S.opt.FL) vaccine is comparable to that generated after four doses, suggesting that three doses are sufficient to elicit robust immune responses. Conversely, four doses of S1.opt pDNA vaccine, containing the S globular head, are required to elicit high levels of neutralizing antibodies. Furthermore, the S.opt.FL pDNA vaccine induces the highest serum levels of interferon (IFN)-γ, a marker for activation of cellular immune responses. Overall, our data show that three doses of S.FL pDNA vaccine elicit potent neutralizing antibody responses, with preclinical data that support the immunogenicity of these COVID-19 vaccine candidates and provide justification for further translational studies.

Delivery, fate and physiological effect of engineered cobalt ferrite nanoparticles in barley (Hordeum vulgare L.).

Cobalt ferrite nanoparticles (CoFe2O4 NPs) have received increasing attention in a widespread application. This work examines the fate and impact of terbium (Tb) substituted CoFe2O4 NPs on the growth, physiological indices, and magnetic character of barley (Hordeum vulgare L.). Sonochemically synthesized NPs were hydroponically applied on barley with changing doses (125-1000 mg/L) at germination and seedling (three weeks) stages. Results revealed a significant reduction in germination rate (∼37% at 1000 mg/L); however, a remarkable growth (∼38-65%) and biomass (∼72-133%) increase were detected at three weeks of exposure (p < 0.05). The elements that make up the NPs (i.e., Tb, Co, and Fe) increased significantly in both root and leaf tissues, indicating the translocation of NPs from the root to leaf. Vibrating-sample magnetometer (VSM) analysis confirmed this finding, where magnetic signals in the root and leaf samples of the control were respectively about 26 and 75 times lower than that of NPs-treated tissues. Also, the accumulation of NPs altered the leaf photoluminescence (PL) behavior, which may have contributed to the biomass increase. Overall, Tb-doped CoFe2O4 NPs translocate from root-to-leaf and enhance plant growth, possibly due to i) incorporation of iron within tissues, and ii) changes in photoluminescence. However, since its effects on other living things are not known yet, its agricultural use and release to nature should be considered well.

Effect of Nigella sativa on general health and immune system in young healthy volunteers; a randomized, placebo-controlled, double-blinded clinical trial.

Nigella sativa ( N. sativa) is traditionally used as an immune enhancer in different communities. The aim of this study was to evaluate the effect of N. sativa on immunity related parameters in young healthy subjects. This study was a double blind, randomized, placebo controlled clinical trial. Fifty-two healthy subjects (48 male and 4 female) 18-25 years old were enrolled in the study. They were randomly divided into four groups; the first received charcoal capsules and served as controls and the other three received 0.5, 1 g, and 2 g of powdered N. sativa capsules, respectively. Two blood samples were obtained from all participant, before initiation of the trial and at the end of the four weeks intervention. One sample was used for routine health screening by evaluating liver and renal functions as well as complete blood count and differential. The second sample was used to measure certain cytokines including; IL-1, IL-4, IL-6, IL-10, and TNF. A third and fourth samples were obtained from the last cohort of subjects before and after treatment; the third was used for measuring immunoglobulins and CD profile and the fourth for evaluating certain gene expressions (INF-γ, NF-κ-B, TNF-α, IL-1ß, IL-13, IL-8, and IL-6). Only 1 g dose of N. sativa produced a significant elevation in total lymphocyte count, CD3+ and CD4+ counts. One gram N. sativa increased the absolute lymphocyte count from 1850±0.24 to 2170±0.26 (p=0.008), CD3+ from 1184.4±75.60 to 1424±114.51 (p=0.009), and CD4+ from 665.6±141.66 to 841±143.36 (p=0.002).  This elevation in T cells was lost by increasing the dose of N. sativa to 2g. The rest of the parameters were not changed significantly in all doses. The results show a promising immunopotentiation effect of N. sativa by elevating helper T cells and the optimum dose for young age group seems to be 1 g.

Uptake, translocation, and physiological effects of hematite (α-Fe2O3) nanoparticles in barley (Hordeum vulgare L.).

There has been a growing concern with the environmental influences of nanomaterials due to recent developments in nanotechnology. This study investigates the impact and fate of hematite nanoparticles (α-Fe2O3 NPs) (∼14 nm in size) on a crop species, barley (Hordeum vulgare L.). For this purpose, hematite NPs (50, 100, 200, and 400 mg/L) were hydroponically applied to barley at germination and seedling stages (three weeks). Inductively coupled plasma mass spectrophotometry (ICP-MS) along with vibrating sample magnetometer (VSM) techniques were used to track the NPs in plant tissues. The effects of NPs on the root cells were observed by scanning electron microscopy (SEM) and confocal microscopy. Results revealed that α-Fe2O3 NPs significantly reduced the germination rate (from 80% in control to 30% in 400 mg/L), as well as chlorophyll (36-39%) and carotenoid (37%) contents. Moreover, the treatment led to a significant decline in the quantum yield of photosystem II (Fv/Fm). Leaf VSM analysis indicated a change in magnetic signal for NPs-treated samples compared with untreated ones, which is mostly attributed to the iron (Fe) ions incorporated within the leaf tissue. Besides, Fe content in the roots and leaf had gradually increased by the increasing doses of NPs, which was confirming NPs' translocation to the aerial parts. Microscopic observations revealed that α-Fe2O3 NPs altered root cell morphology and led to the injury of cell membranes. This study, in the light of our findings, shows that α-Fe2O3 NPs (∼14 nm in size) are taken up by the roots of the barley plants, and migrate to the plant leaves. Besides, NPs are phytotoxic for barley as they inhibit germination and pigment biosynthesis. This inhibition is probably due to the injury of the cell membranes in the roots. Therefore, the use of hematite NPs in agriculture and thereby their environmental diffusion must be addressed carefully.

Engineered magnetic nanoparticles enhance chlorophyll content and growth of barley through the induction of photosystem genes.

This study investigates the impact of an engineered magnetic nanoparticle (MNP) on a crop plant. For this purpose, a sonochemical synthetic approach was utilized in order to dope magnetic elements (Co and Nd) into technologically important iron oxide NPs. After being characterized by using TEM, SEM, and XRD instruments, the MNPs were hydroponically applied to barley plants with varying doses (from 125 to 1000 mg/L) both in germination (4 days) and early growing stages (3 weeks). Physiological responses, as well as expression of photosystem marker genes, were assessed. Compared to the untreated control, MNP treatment enhanced germination rate (~ 31%), tissue growth (8% in roots, 16% in shoots), biomass (~ 21%), and chlorophyll (a, b) (~ 20%), and carotenoids (~ 22%) pigments. In general, plants showed the highest growth enhancement at 125 or 250 mg/L treatment. However, higher doses diminished the growth indices. Compared to the control, the catalase activity was significantly reduced in the leaves (~ 33%, p < 0.005) but stimulated in the roots (~ 46%, p < 0.005). All tested photosystem marker genes (BCA, psbA, and psaA) were overexpressed in MNP-treated leaves than non-treated control. Moreover, the gene expressions were found to be proportionally increased with increasing MNP doses, indicating a positive correlation between MNPs and the photosynthetic machinery, which could contribute to the enhancement of plant growth.

Size effect of iron (III) oxide nanomaterials on the growth, and their uptake and translocation in common wheat (Triticum aestivum L.).

Nanomaterials (NMs) have emerged in the last decades and are used in many disciplines such as industry, material sciences, biomedicine, biotechnology, bioenergy, and agriculture. The size of the NMs is a critical factor that affects NMs' integration and transfer into the biological systems. Therefore, this study aims at investigating the effect of NMs-size on i) plant growth and physiology, and ii) NMs uptake and translocation in plant tissues. For these purposes, iron (III) oxide (Fe2O3) NMs with varied sizes, 8-10, 20-40, and 30-50 nm, have been applied to wheat plants in a hydroponic system. Results showed that Fe2O3 NMs enhanced root length, plant height, biomass, and chlorophyll content of wheat. Confocal microscopy analysis indicated that Fe2O3 NMs cause injury in root-tip cells without a visible toxic symptom. Vibrating sample magnetometer (VSM), and inductively coupled plasma-mass spectroscopy (ICP-MS) analyses of leaf tissues revealed that all tested NMs were up taken by wheat plant and translocated to the leaves. Iron content was found to be dramatically increased in NMs-treated plant tissues, which possibly contributed to the growth enhancement. Experiments confirmed that Fe2O3 NMs with 20-40 nm size is much more efficient in plant growth compared to those with 8-10 and 30-50 nm size. Overall, Fe2O3 NMs with 20-40 nm in size could be proposed as a nano-fertilizer for agricultural applications. On the other hand, the translocation of NMs in the wheat plant requires further investigation of their effects on the end users.

Impact of calcium and magnesium substituted strontium nano-hexaferrite on mineral uptake, magnetic character, and physiology of barley (Hordeum vulgare L.).

In this study, calcium and magnesium substituted strontium nano-hexaferrites (Sr0.96Mg0.02Ca0.02Fe12O19, SrMgCa nano-HF) were synthesized by the sol-gel auto-combustion method and their impact on the nutrient uptake, magnetic character and physiology of barley (Hordeum vulgare L.), a crop plant, was investigated. Structural, microstructural, and magnetic properties of nano-HF were evaluated by using vibrating sample magnetometry (VSM), X-ray diffraction (XRD), scanning electron microscopy (SEM) along with energy-dispersive X-ray (EDX) and elemental mapping techniques. Plants were hydroponically exposed to nano-HF (ranging from 125 to 1000 mg/L) for three weeks. Results showed that the SrMgCa nano-HF application enhanced germination rate (about 20%), tissue growth (about 38%), biomass (about 20%), soluble protein content (about 41%), and chlorophyll pigments (about 33-42%) when compared to the untreated control. In general, the plants showed the highest growth achievement at 125 or 250 mg/L of nano-HF treatment. However, higher doses diminished the growth parameters. Element concentrations and magnetic behavior analyses of plant parts proved that SrMgCa nano-HF with a size of 42.4 nm are up-taken by the plant roots and lead to increase in iron, calcium, magnesium, and strontium contents of leaves, which were about 20, 18, 3, and 60 times higher in 500 mg/L nano-HF-treated leaves than those of control, respectively. Overall, this study shows for the first time that the four elements have been internalized into the plant body through the application of substituted nano-HF. These findings suggest that mineral-substituted nanoparticles can be incorporated into plant breeding programs for the i) enhancement of seed germination and ii) treatment of plants by fighting with mineral deficiencies.