DNA damage through endogenous and environmental toxicants is a constant threat to both a human's ability to pass on intact genetic information to its offspring as well as in somatic cells for its own survival. To counter these threats posed by DNA damage, cells have evolved a series of highly choreographed mechanisms-collectively defined as the DNA-damage response (DDR)-to sense DNA lesions, signal their presence, and mediate their repair. Thus, regular DDR signaling cascades are vital to prevent the initiation and progression of many human diseases including cancer. Consequently, quantitative assessment of DNA damage and response became an important biomarker for assessment of human health and disease risk in biomonitoring studies. However, most quantitative DNA damage biomarker techniques require dissolution of the nuclear architecture and hence loss of spatial information. Laser scanning confocal immunofluorescence microscopy (LSCIM) of three-dimensionally preserved nuclei can be, quantitative and maintain the spatial information. Here we describe the experimental protocols to quantify individual key events of the DDR cascade in three-dimensionally preserved nuclei by LSCIM with high resolution, using the simultaneous detection of Rad50 as well as phosphorylated H2AX and ATM and in somatic and germ cells as an example.

Human biomonitoring studies aim to identify potential exposures to environmental, occupational, or lifestyle toxicants in human populations and are commonly used by public health decision makers to predict disease risk. The Comet assay measures changes in genomic stability and is one of the most reliable biomarkers to indicate early biological effects, and therefore accepted by various governmental regulatory agencies. The appeal of the Comet assay lies in its relative simplicity, rapidity, sensitivity, and economic efficiency. Furthermore, the assay is known for its broad versatility, as it can be applied to virtually any human cell and easily adapted in order to detect particular biomarkers of interest, such as DNA repair capacity or single- and double-strand breaks. In a standard experiment, isolated single cells are first embedded in agarose, and then lysed in high-salt solutions in order to remove all cellular contents except the DNA attached to a nuclear scaffold. Subsequent electrophoresis results in accumulation of undamaged DNA sequences at the proximity of the nuclear scaffold, while damaged sequences migrate towards the anode. When visualized with fluorochromes, these migrated DNA fragments resemble a comet tail and can be quantified for their intensity and shape according to internationally drafted guidelines.

Psoriasis is an immune-mediated disease affecting the skin and joints. biologic treatments are often used in moderate to severe cases, administered as a monotherapy or in combination with other treatments. Liposomes are versatile and can enhance drug retention, reducing systemic side effects, and are used for therapy and research. This research study aimed to investigate the Geno toxicity of Secukinumab, an ideal biologic treatment for psoriasis (a human IgG1k antibody, anti-IL17A), in bulk and liposome nanoparticles on the lymphocytes of psoriatic patients in comparison with healthy persons. Geno toxicity of Secukinumab in bulk and liposomal form was evaluated and compared by using the Comet and micronucleus assays. From assays, it was demonstrated that Secukinumab in both forms did not exhibit Geno toxicity and reduced DNA damage following the treatment of psoriatic patient lymphocytes and healthy individual’s lymphocytes with Secukinumab bulk and liposomes format. Secukinumab used (2.1 and 2.8 µg/mL) with two different concentrations, and effectively decreased DNA damage induced by H2O2 in both groups to almost the negative control level. Secukinumab bulk and liposome form markedly reduced the H2O2-induced damage and proficiently diminished its adverse effects both in the Comet (p<0.0001) and micronucleus as-says (p<0.01). Overall, Secukinumab in both forms showed anti-genotoxic and protective effects by expressing its potential to reduce DNA damage produced by oxidative stress and it was observed that it would not induce any further damage in the lymphocytes of healthy individuals and patients.

Abstract Background Background: Free radicals and reduced antioxidant levels lead to oxidative stress, which plays a role in causing Inflammatory Bowel Disease (IBD). IBD is a chronic gastrointestinal autoimmune condition that involves an abnormal immune response. This includes Crohn's disease (CD) and ulcerative colitis (UC). In the previous studies, we demonstrated the antioxidant effect of Inonotus obliquus (IO) and flavonoids such as quercetin and epicatechin on lymphocytes from IBD patients. In vitro studies have shown that Chaga extract is effective in reducing oxidative stress in lymphocytes of both healthy individuals and those with IBD. This suggests that Chaga extract may be a useful supplement for anyone looking to inhibit oxidative stress. Methods Methods: In our current study, Cord Blood Stem Cells -derived Exosomes (CBSC Exo) and their synthesised miRNAs were investigated as antioxidant/anti-inflammatory elements on lymphocytes and 3D intestinal epithelial model. There were seven miRNAs found in CBSCs Exo, and among them were two miRNAs that were identified as novel. The Peripheral Blood Monocyte Cells (PBMCs) from IBD patients and healthy individuals after challenging with H2O2 or Interleukin 6 were treated with CBSCs Exo, transfected with miRNA novels or both. The treatment method also was applied to the EpiltestinalTK 3D model from Mattek. Various techniques were utilised, including the Comet assay, Fast Microplate DNA damage assay, and CCK8. Results The findings of the study indicate that the administration of CBSC-derived Exo, Let-7c-mimic miRNA, Let7bSb miRNA, as well as Novel 1 and Novel 2 miRNAs on the 3D model intestine, significantly decreased the level of DNA damage when compared to the positive control H2O2 and IL6, as well as the untreated samples. Conclusion Conclusions: In conclusion, certain natural substances like flavonoids, Inonotus obliquus, and exosomes derived from CBSCs may have an advantage in decreasing significantly oxidative stress and DNA damage in IBD Peripheral Blood Mononuclear Cells (PBMCs) and inflamed intestinal 3D models.

Abstract Background The current treatments for IBD frequently do not provide sufficient control over the disease, warranting the investigation of alternative therapeutic options with minimal risk of side effects. In IBD, the overgeneration and insufficient removal of reactive oxygen species (ROS) leads to oxidative stress. Reduced glutathione (GSH) and n-acetylcysteine (NAC) are antioxidants which may possess therapeutic potential in IBD, by modulating endogenous mechanisms that decrease ROS production or increase antioxidant enzymes. Additonally, exosomes from human umbilical cord blood derived mesenchymal stem cells (hucMSCs-exo) may be used to treat IBD. These are a subset of extracellular nanosized membrane vesicles which participate in intercellular communication by delivering their contents, such as functional miRNAs to recipient cells, thereby influencing the physiological and pathological processes in various diseases. The therapeutic potential of exosomes can be enhanced through the overexpression of the let-7 family of miRNAs. Methods The EpiIntestinal tissue model is a physiologically relevant predictor of drug-induced GI toxicity. EpiIntestinal cells and lymphocytes from 3 IBD patients were transfected with 50 μL miRNA mimics and inhibitors, and treated with 50μL hucMSC-exo, and 50 μL 1mM GSH and NAC. Next, in comparison to negative controls, single-stranded DNA damage was assessed in peripheral blood mononuclear cells (PBMCs) from IBD patients using the fast microplate DNA damage assay, and double-stranded DNA damage was assessed in EpiIntestinal cells using the alkaline Comet assay. In addition, the viability of healthy and IBD lymphocytes was assessed following treatment with selected concentrations of hucMSC-exo, GSH and NAC. Results Double-stranded DNA damage was significantly reduced in EpiIntestinal cells compared to the negative control following treatment with hucMSC-exo, GSH, NAC, and miRNA mimics for let-7a-5p, -7b-5p, -7c-3p, -7d-3p and -7d-5p. In addition, single-stranded DNA damage was reduced in IBD PBMCs compared to the negative control following treatment with hucMSC-exo, and let-7b-5p and -7c-3p mimics, but was increased following treatment with inhibitors for let-7b-5p, -7c-5p and two novel miRNAs. Additionally, GSH, NAC and hucMSC-exo increased cell viability in treated versus untreated lymphocytes from healthy individuals and IBD patients. Conclusion Selected concentrations of HucMSC-exo, GSH, NAC and/or let-7 miRNA mimics reduced DNA damage and increased cell viability in treated versus untreated cells which may reduce the risk of colorectal cancer in IBD patients. Acknowledgements Our gratitude goes to the MatTek Team for generously granting us a SMI-100-FT EpiIntestinal kit in support of this research.

Abstract Regulatory T (Treg) cells expressing FOXP3 play a critical role in suppressing immune responses against anti-tumor immune responses. However, their presence in tumor tissues is often associated with poor prognosis. Cord blood stem cell-derived exosome vesicles (CBSC-EV) constitute a valuable source of lipids, microRNAs, and proteins, that are essential for cell-to-cell communication and provide therapeutic benefits. This study aimed to investigate the contents of CBSC-EV, identifying several known miRNAs (from Let7 and Let5 families) and two unknown miRNAs. Healthy lymphocytes, fibroblasts, and CHL1 amelanotic melanoma cell lines were transfected with these two novel miRNA inhibitors, and the changes before and after treatment compared to CBSC-EV and H2O2 were examined. Following treatment, the cells underwent a 24-hour incubation period, during which the level of cytotoxicity was assessed using CCK8, and the degree of DNA damage incurred was quantified with the Fast Microplate DNA damage assay. To further investigate the effects of treatment, RNA sequencing was conducted on both healthy and cancerous cells. The results demonstrated that CBSC-EV and novel miRNAs induced cytotoxicity in cancer cells, while in healthy cells, the DNA damage was repaired and cell viability was significantly increased (p<0.001). RNA sequencing revealed that following treatment with CBSC-EV and both CBSC-EV+N1 and N2 inhibitors in malignant melanoma cells, the expression of genes involved in the cytokine-mediated signaling pathway, cytokine-cytokine interaction, and cytokine activity were enhanced. In healthy cells treated with CBSC-EV and novel miRNA inhibitor transfection, the MT-CO1 gene was upregulated, contributing to cytochrome-c oxidase activity. This study presents evidence of the potential use of novel miRNAs as genetic material for cancer therapeutics in the future. Additionally, the findings suggest that these miRNAs could be used as an immune system modulator against melanoma. These findings demonstrate that miRNAs can effectively suppress resistance to anticancer cytotoxic therapy, which is a common feature of cancer cells. This suggests that miRNAs can potentially enhance current cancer therapies. Citation Format: Mojgan Mojgan Najafzadeh, Adi Baumgartner, Shohreh Jafarinejad, Zahra Karimi, Mohammad Isreb, Pouria Akhbari, Nader Ghaderi, Farshid Sefat, Saeed Heidari Keshel, Parisa Naeem, Rojan Ghaderi, Jacobo Elis Gomez, Diana Anderson, Andrew Wright. The exosomes from cord blood stem cells, containing novel miRNAs, induce apoptosis in melanoma cells and enhance T cell anticancer function [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2783.

This study aimed to investigate the potential of Cord Blood Stem Cell-derived exosome vesicles (CBSC-EVs) in regulating cancer growth and preventing cellular damage. The study aimed to examine the effects of CBSC-EVs on CHL1 cancer cells and healthy fibroblasts. The study results reveal that CBSC-EVs have an anti-cancer/antioxidant effect on CHL1 cancer cells without inducing any toxic effects on healthy lymphocytes. Further analysis of CBSC-EVs revealed the presence of seven miRNAs (noncoding RNA molecules), including three from the Let 7 family, two from the Let 5 family, and two novel miRNAs. The function of the novel miRNAs was the main focus of the study. To assess the efficacy of CBSC-EVs, CHL1 cells (a type of malignant melanoma cell line) and human fibroblasts were grouped into different treatments. One group of those cell lines were treated with CBSC-EVs, and in the second group, the cells were transfected with either miRNA inhibitor Novel 1 (miRNA IN1) plus CBSC-EVs or miRNA inhibitor Novel 2 (miRNA IN2) and CBSC-EVs to knock down those novel miRNAs in the CBSC-EVs. Therefore, by comparing these groups, the function of novel miRNAs is more understandable. In this study, the Comet assay was employed to evaluate cancer cells' DNA damage and CCK8 to measure cell viability. The findings revealed that CBSC-EVs caused an increase in DNA damage in CHL1 cells compared to cells without treatment (p

The testis is a complex organ that serves two important functions, namely, synthesis of steroids and production of spermatozoa to attain fertility, which are coordinated by gonadotrophins and various locally synthesized factors. The function of oestrogen in the male reproductive system has been a matter of ongoing debate. The testes produce significant amounts of oestrogenic hormones through the enzyme aromatase and oestrogen receptors (ERs), namely ERα (ESR1) and ERβ (ESR2). These receptors are selectively expressed in the cells of the testis as well as the epididymal epithelium. The importance of oestrogen signalling in male fertility is indicated by the adverse effects of selected oestrogen-like compounds, and their interaction with oestrogen receptors was proven to cause pathologies. The aim of this chapter is to review the current knowledge on oestrogen signalling and adverse effects on spermatogenesis and how they are linked to the induction of germ cell DNA damage and apoptosis.

Pembrolizumab has shown significant anticancer effects against various human cancers. The present study investigated the effects of pembrolizumab liposome and nano (naked) forms in treated lymphocytes from head and neck squamous cell carcinoma (HNSCC) patients compared to healthy individuals. The level of oxidative DNA damage induced by hydrogen peroxide (H2O2) was also investigated. A concentration of 10 mu g/ml of pembrolizumab liposome was used to treat the lymphocytes in the Comet and micronucleus assays based on the preliminary dosage optimization tests. To determine the cellular pathways involved in the protective role of pembrolizumab against H2O2, several proteins involved in apoptosis (P53, P21 and Bcl-2) were assessed. Pembrolizumab significantly reduced DNA damage and decreased the number of micronuclei in lymphocytes from HNSCC patients (p < 0.01) compared with healthy individuals. The 10 g/ml of pembrolizumab liposome significantly reduced the oxidative stress induced by H2O2 and was effective in healthy and HNSCC groups using the Comet and micronucleus assays (p < 0.001). To our knowledge, this is the first report of pembrolizumab in liposome and naked forms exhibiting a protective effect on DNA damage in the treatment of HNSCC patients.

Psoriasis is a multi-factorial immune-mediated disease that affects approximately 2% of the UK population and 2.5% worldwide. The condition can impact multiple organs, including the skin and joints. In moderate to severe cases, systemic and topical medications may not provide adequate relief, and as a result, biologic treatments are often administered as a monotherapy or in combination with other treatments. While biologic treatments are more effective than systemic and topical medications in treating psoriasis, patients require close supervision and monitoring while receiving this type of treatment. Liposomes are highly versatile and can be employed for therapy and research applications. They enhance drug retention at the site of administration, thereby reducing systemic side effects. Our study was the first to investigate the genotoxicity of secukinumab, an ideal biologic treatment for psoriasis (a human IgG1k antibody, anti-IL17A), in bulk and liposome nanoparticles on the lymphocytes of psoriasis patients compared to healthy individuals. We evaluated and com-pared the genotoxicity of secukinumab in bulk and liposomal form using the Comet and micronucleus assays. Both assays demonstrated that secukinumab in both forms did not ex-hibit genotoxicity and reduced DNA damage following treatment of the lymphocytes from healthy individuals and psoriatic patients with secukinumab bulk and liposomes format. Both concentrations of secukinumab used (2.1 and 2.8 µg/mL) effectively decreased H2O2-induced DNA damage in both groups to nearly the level of the negative control. Secukinumab bulk and liposome form significantly decreased H2O2-induced damage and efficiently attenuated its adverse effects both in the Comet (p<0.0001) and micronucleus assays (p<0.01). Overall, secukinumab in both forms exhibited protective and an-ti-genotoxic effects by demonstrating its potential to reduce DNA damage caused by oxida-tive stress and by not inducing any further damage in the lymphocytes of either healthy in-dividuals or patients.

Globally, lung cancer affected 2.2 million individuals and caused 1.8 million deaths in 2021. Lung cancer is caused by smoking, genetics and other factors. IFN-γ has anticancer activity. However, the mechanism by which IFN-γ has an effect on lung cancer is not fully understood. The present study aimed to assess the effect of IFN-γ on the peripheral lymphocytes of patients with lung cancer compared with healthy controls. The efficacy of IFN-γ against oxidative stress was assessed using a comet repair assay and the effects of IFN-γ on p53, PARP1 and OGG1 genes and protein levels in lymphocytes was evaluated by RT-qPCR and western blotting. DNA damage was significantly reduced in the lymphocytes of patients treated with IFN-γ. However, there was no effect in the cells of healthy individuals after treatment with naked IFN-γ [IFN-γ (N)] and liposomal IFN-γ [IFN-γ (L)]. Following treatment with IFN-γ (N) and IFN-γ (L), the p53, PARP1 and OGG1 protein and gene expression levels were significantly increased (P<0.001). It has been suggested that IFN-γ may induce p53-mediated cell cycle arrest and DNA repair in patients. These findings supported the idea that IFN-γ (N) and IFN-γ (L) may serve a significant role in the treatment of lung cancer, via cell cycle arrest of cancer cells and repair mechanisms.

The Comet assay, a highly sensitive method for detecting DNA damage, is used to measure DNA breaks and alkali-labile lesions in eukaryotic cells. This study outlines the application of whole blood in the alkaline gel electrophoresis method. A total of 1,200 blood samples from various individuals were analysed, comprising 300 samples from healthy individuals, 300 from suspected or pre-cancerous individuals, and 600 from cancer patients. Each sample was divided into two identical volumes in separate falcon tubes. The blood samples were prepared and preserved by adding an equal amount of RPMI medium and 10% DMSO. Analysis of the data using Student’s t-Test revealed a p-value of 0.59 for Olive tail moment (OTM) and 0.16 for % tail DNA, indicating no statistically significant differences between the two methods, whether with or without treatment. In conclusion, employing whole blood instead of isolated lymphocytes not only saves time but also maintains a high level of sensitivity, requiring less than 20 µL of blood from each individual.

The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA-DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility.

This study compared the expression of TP53 in lymphocytes from malignant melanoma (MM) patients with positive sentinel nodes to healthy controls (HCs) following exposure to various doses of UVA radiation. The Lymphocyte Genome Sensitivity (LGS) assay indicated significant differences in DNA damage in lymphocytes between MM patients and HCs. qPCR data demonstrated an overall 3.4-fold increase in TP53 expression in lymphocytes from MM patients compared to healthy controls, following treatment with 0.5 mW/cm 2 UVA radiation. Western blotting confirmed that p53 expression was increased in MM lymphocytes following UVA exposure compared to healthy individuals. Genome transcriptome profiling data displayed differences in gene expression between UVA-treated lymphocytes from MM patients and HCs. Peripheral lymphocytes from MM patients are more susceptible to the genotoxic effects of UVA compared to healthy individuals. Our previous studies showed that UVA exposure of various intensities caused significant differences in the levels of DNA damage between lymphocytes from cancer patients compared to HCs through the LGS assay. The present study’s results provide further credibility to the LGS assay as a screening test for cancer detection. Peripheral lymphocytes could be a promising blood biopsy biomarker for staging of carcinomas and prevention of carcinoma progression at early stages.

Abstract This study aimed to evaluate the expression of the P53 gene following exposure to varying doses of UVA radiation, using lymphocytes as surrogates. Lymphocytes from malignant melanoma (MM) patients (n = 20) with positive sentinel nodes were compared to healthy controls (HC)(volunteers) (n = 20). These samples were processed by Comet assay following the Lymphocyte Genome Sensitivity (LGS) test, quantitative real-time Polymerase Chain Reaction (qPCR), western blotting and whole genome transcriptome profiling. LGS test evaluates the level of alterations in lymphocytes resulting from continuous exposure to various physical and chemical insults in the blood, promoting DNA damage, ultimately leading to oxidative stress. It is believed that in cancer, the circulatory tumour cells, exosomes and cytokines impact peripheral lymphocytes. The Comet assay performed within the LGS test indicated a significant difference between the lymphocytes from two groups of HC and MM patients. The qPCR data demonstrated an overall 43.8-fold increase in TP53 gene expression in lymphocytes from MM patients after treatment with 0.2mW/cm2 UVA intensity radiation, compared to healthy and untreated controls. Western blotting was used to confirm gene expression. The p53 protein expression was significantly increased in MM lymphocytes after UVA exposure compared to healthy individuals (p-value < 0.05). The genome transcriptome profiling data also displayed differences in gene expression between the UV-treated lymphocytes from healthy groups as compared to melanoma samples. Nine out of the 23 (~ 40%) genes displaying differences in gene expression were mitochondrial genes, which were increased in lymphocytes from MM compared to HCs. The genes that play an important role in oxidative phosphorylation, such as MT-CYB, MT-CO2, MT-ND2, MT-ND6 and MTRNR2L12, were upregulated in lymphocytes from MM patients compared to HCs. The down-regulated genes in lymphocytes from MM, such as MYH9, RN7SL2, ACTB, AHNAK and FLNA, are related to cell structure, migration and tumour metastasis. Peripheral lymphocytes from MM patients are more sensitive and susceptible to the genotoxic effects of UVA compared to healthy individuals. Our previous studies showed that UVA exposure in various intensities distinguishes differences in the level of DNA damage between lymphocytes from cancer patients compared to HCs through the LGS test. The current results provide further credibility to the LGS assay as a screening test for detecting cancer. This feature could be a promising blood biopsy biomarker for staging and preventing carcinomas at early stages.

Incidence of Malignant Melanoma has become the 5th in the UK. To date, the major anticancer therapeutics include cell therapy, immunotherapy, gene therapy and nanotechnology-based strategies. Recently, extracellular vesicles, especially exosomes, have been highlighted for their therapeutic benefits in numerous chronic diseases. Exosomes display multifunctional properties, including inhibition of cancer cell proliferation and initiation of apoptosis. In the present in vitro study, the antitumour effect of cord blood stem cell (CBSC)-derived exosomes was confirmed by the CCK-8 assay (p < 0.05) on CHL-1 melanoma cells and improve the repair mechanism on lymphocytes from melanoma patients. Importantly, no significant effect was observed in healthy lymphocytes when treated with the exosome concentrations at 24, 48 and 72 h. Comet assay results (OTM and %Tail DNA) demonstrated that the optimal exosome concentration showed a significant impact (p < 0.05) in lymphocytes from melanoma patients whilst causing no significant DNA damage in lymphocytes of healthy volunteers was 300 μg/ml. Similarly, the Comet assay results depicted significant DNA damage in a melanoma cell line (CHL-1 cells) treated with CBSC-derived exosomes, both the cytotoxicity of CHL-1 cells treated with CBSC-derived exosomes exhibited a significant time-dependent decrease in cell survival. Sequencing analysis of CBSC exosomes showed the presence of the let-7 family of miRNAs, including let-7a-5p, let-7b-5p, let-7c-5p, let-7d-3p, let-7d-5p and two novel miRNAs. The potency of CBSC exosomes in inhibiting cancer progression in lymphocytes from melanoma patients and CHL-1 cells whilst causing no harm to the healthy lymphocytes makes it a potential candidate as an anticancer therapy.

An acute respiratory disease caused by the 2019 novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), otherwise known as coronavirus disease 2019 (COVID-19), recently emerged in Wuhan, Hubei Province, China, and has since spread rapidly across the globe. As of 30 January 2020, the COVID-19 epidemic was officially declared as a public health emergency of international concern by the World Health Organization (WHO). Coronaviruses are enveloped positive-sense RNA viruses that can cause a variety of diseases in both birds and mammals. SARS-CoV-2 is a novel strain that has not been previously identified in humans. However, it is the third introduction of a highly pathogenic coronavirus into the human population since SARS-CoV-1 was identified in 2002 and Middle East respiratory syndrome (MERS-CoV), which was first identified in 2012. To date, no effective or specific treatment has been identified for SARS-CoV-2; despite this, certain candidates have shown great efficacy in viral inhibition of the disease. Natural substances have previously exhibited anti-viral and anti-inflammatory activity. Thus, the possibilities of natural substances as effective treatments against SARS-CoV-2 may seem promising. This chapter aims to provide a comprehensive overview of the possible use of natural products against SARS-CoV-2 by evaluating their anti-viral and anti-inflammatory effects, which have been assessed previously in laboratory conditions.

Reproductive health embraces a range of factors and processes, comprising wellbeing and physical health, and is important in maintaining the quality and quantity of future generations. The current pandemic status of the novel coronavirus disease 2019 (COVID-19) outbreak on reproductive health is of a significant public concern within the epidemic. There is only limited scientific evidence available to identify the effects of COVID-19 on reproductive health, principally consisting of reported outcomes of the infection during pregnancy and published cases of COVID-19 occurring during pregnancy. Most women admitted to hospital have been in the late second or third trimesters of pregnancy and from black or other ethnic minority groups. However, questions arise concerning the continuation of pregnancy, the possibility of virus transmission through the placenta, isolation of the newborn after birth, and breastfeeding. The health implications of the spread of COVID-19 from the initial site of infection to the female reproductive organs in pregnant women are the main focus of this review; male reproductive effects are also considered.