Green synthesis of silver doped zinc oxide/magnesium oxide nanocomposite for waste water treatment and examination of their cytotoxicity properties.

This research attempted to prepare silver-doped zinc oxide/magnesium oxide nanocomposite (Ag-doped ZnO/MgO-NCP) using Mentha pulegium plant extract. The synthesized NCP was investigated by X-ray diffraction analysis (XRD), Fourier Transform Infrared (FT-IR), Field Emission Scanning Electron Microscope (FESEM), Energy dispersive X-ray spectroscopy (EDX), Mapping, and UV-Visible analyses. The XRD data displayed cubic crystal structures for silver & magnesium oxide and a hexagonal framework for zinc oxide. Also, FESEM and PSA images of NCP pointed out, that the average size of the spherical morphology is about 10-16 nm, while the analysis of EDX confirmed the attendance of Zn, Mg, Ag, and O elements. Under UVA light, we tested the photocatalytic activity of NCP to the degradation of Methylene blue (MB) and Rhodamine B (RhB) dyes in various temperatures (400, 500, and 600 °C). The results of the photocatalytic test displayed that the degradation percentage of MB dye in pH = 9, nanocomposite amount ∼30 mg, and dye concentration ∼1 × 10 -5 M was about 98 %. We also evaluated the cytotoxicity of nanocomposite on cancer CT-26 cell line through the MTT method and obtained an IC50 value of 250 µg/mL.

In silico approaches to investigate enzyme immobilization: a comprehensive systematic review.

Enzymes are popular catalysts with many applications, especially in industry. Biocatalyst usage on a large scale is facing some limitations, such as low operational stability, low recyclability, and high enzyme cost. Enzyme immobilization is a beneficial strategy to solve these problems. Bioinformatics tools can often correctly predict immobilization outcomes, resulting in a cost-effective experimental phase with the least time consumed. This study provides an overview of in silico methods predicting immobilization processes via a comprehensive systematic review of published articles till 11 December 2022. It also mentions the strengths and weaknesses of the processes and explains the computational analyses in each method that are required for immobilization assessment. In this regard, Web of Science and Scopus databases were screened to gain relevant publications. After screening the gathered documents (n = 3873), 60 articles were selected for the review. The selected papers have applied in silico procedures including only molecular dynamics (MD) simulations (n = 20), parallel tempering Monte Carlo (PTMC) and MD simulations (n = 3), MD and docking (n = 1), density functional theory (DFT) and MD (n = 1), only docking (n = 11), metal ion binding site prediction (MIB) server and docking (n = 2), docking and DFT (n = 1), docking and analysis of enzyme surfaces (n = 1), only DFT (n = 1), only MIB server (n = 2), analysis of an enzyme structure and surface (n = 12), rational design of immobilized derivatives (RDID) software (n = 3), and dissipative particle dynamics (DPD; n = 2). In most included studies (n = 51), enzyme immobilization was investigated experimentally in addition to in silico evaluation.

Biosynthesis of Gold Nanoparticles Using Quince Seed Water Extract and Investigation of Their Anticancer Effect Against Cancer Cell Lines.

In this study, gold nanoparticles (Au-NPs) were synthesized using HAuCl4 and quince seed mucilage (QSM) extract, which was characterized by conventional methods including Fourier transforms electron microscopy (FTIR), UV-Visible spectroscopy (UV-Vis), Field emission electron microscopy (FESEM), Transmission electron microscopy (TEM), Dynamic light spectroscopy (DLS), and Zeta-potential. The QSM acted as reductant and stabilizing agents simultaneously. The NP's anticancer activity was also investigated against osteosarcoma cell lines (MG-63), which showed an IC50 of [Formula: see text]/mL.

Development of a Novel Sulfur Quantum Dots: Synthesis, 99mTc Radiolabeling, and Biodistribution.

Sulfur quantum dots (SQDs) as free heavy metal element quantum dots have promising applications in diagnosis and therapy; however, SQDs' in vivo biodistribution has not been studied. In the current study, SQDs were synthesized directly from cheap sublimated sulfur powder via a one-pot solvothermal method, and sucrose was used as a stabilizer to enhance stability and biocompatibility. The as-obtained SQDs with an average size of 4.6 nm exhibited great water dispersity, highly favorable quantum yield (21.5%), and uniformly spherical shape which were confirmed by UV-Vis, fluorescence spectrophotometer, TEM, and FESEM/EDX/PSA analyses. Moreover, the as-synthesized SQDs had very low cytotoxicity based on cancer (C26) and normal (L929) cell lines via MTT assay. And also, SQDs were radio-labeled directly by Technetium-99m (99mTc), which had good stability ranging from 86 to 99% in PBS and human serum. The SQDs' cell uptake on C26 and L929 cell lines demonstrated that cancer cells had more uptake than normal cells by increasing concentrations. Moreover, SQDs' in vivo biodistribution results displayed high kidney dose accumulation and rapid renal clearance, making them suitable for imaging and therapeutic applications.

Zn(II) porphyrin-encapsulated MIL-101 for photodynamic therapy of breast cancer cells.

The photodynamic treatment is a non-aggressive and clinically accepted procedure for removing selected cancer cells with the activation of a photosensitizer agent at a specific light. In this study, the zinc porphyrin (Zn[TPP]) was prepared and encapsulated into the MIL-101 (Zn[TPP]@MIL-101). It was used in photodynamic therapy (PDT) against MCF-7 breast cancer cells under a red light-emitting diode. The structure, morphology, surface area, and compositional changes were investigated using conventional characterization methods including FTIR, FESEM, EDX, and BET analyses. The MTT assay was performed under light and dark conditions to explore the ability of Zn[TPP]@MIL-101 in PDT. The results have demonstrated the IC50 of 14.3 and 81.6 mg/mL for light and dark groups, respectively. As the IC50 revealed, the Zn[TPP]@MIL-101 could efficiently eradicate cancer cells using PDT.

Design and preparation of amino-functionalized core-shell magnetic nanoparticles for photocatalytic application and investigation of cytotoxicity effects.

The goal of the current paper was a synthesis of Amino-functionalized Fe3O4@SiO2 core-shell magnetic nanoparticles as a unique efficient photocatalyst for removing organic dyes from aqueous environments. The magnetic Fe3O4@SiO2 core-shell was produced by a silica source to avoid aggregation by the co-precipitation method. Next, functionalized by using 3-Aminopropyltriethoxysilane (APTES) via a post-synthesis link. The chemical structure, magnetic properties, and shape of the manufactured photocatalyst (Fe3O4@SiO2-NH2) were described by XRD, VSM, FT-IR, FESEM, EDAX, and DLS/Zeta potential analyses. The XRD findings approved the successful synthesis of nanoparticles. The photocatalytic activity of Fe3O4@SiO2-NH2 nanoparticles was examined for MB degradation and the degradation performance was about 90% in the optimum conditions. Also, the cytotoxicity of Fe3O4, Fe3O4@SiO2 core-shell, and Fe3O4@SiO2-NH2 nanoparticles was examined on CT-26 cells using an MTT assay, the finding has shown that nanoparticles can be used for inhibiting cancer cells.

Green synthesis of CaCO3 nanoparticles for photocatalysis and cytotoxicity.

In this study, Gum Arabic natural polymer was used to green synthesize calcium carbonate nanoparticles (CaCO3-NPs). Synthesized CaCO3-NPs were identified using various analyses such as FTIR, XRD, FESEM, EDX, and UV-Vis. The UV spectrum of nanoparticles showed an absorption band at a wavelength of 320 nm. FTIR analysis also confirmed the synthesis of nanoparticles. XRD studies showed that CaCO3-NPs have a rhombohedral crystalline structure with space group R-3c and an average size of about 42 nm. FESEM images showed that CaCO3-NPs have cubic morphology and EDX results confirmed the presence of carbon, calcium, and oxygen elements. The synthesized CaCO3-NPs showed good photocatalytic activity to methylene blue (MB) dye degradation, which percentage degradation was 93% after 120 min. Also, the cytotoxicity of CaCO3-NPs has been examined on the normal L929 and cancer CT26 cell lines and the IC50 value was about 250 µg/mL for cancer cells.

Green Synthesis of Silver Nanoparticles Using Salvadora persica and Caccinia macranthera Extracts: Cytotoxicity Analysis and Antimicrobial Activity Against Antibiotic-Resistant Bacteria.

Silver nanoparticles (AgNPs) have gained great interest because of their specific and distinct properties. Chemically synthesized AgNPs (cAgNPs) are often unsuitable for medical applications due to requiring toxic and hazardous solvents. Thus, green synthesis of AgNPs (gAgNPs) using safe and nontoxic substances has attracted particular focus. The current study investigated the potential of Salvadora persica and Caccinia macranthera extracts in the synthesis of CmNPs and SpNPs, respectively. Aqueous extracts of Salvadora persica and Caccinia macranthera were prepared and taken as reducing and stabilizing agents through gAgNPs synthesis. The antimicrobial effects of gAgNPs against susceptible and antibiotic-resistant bacterial strains and their toxicity effects on L929 fibroblast normal cells were evaluated. TEM images and particle size distribution analysis showed that the CmNPs and SpNPs have average sizes of 14.8 nm and 39.4 nm, respectively. The XRD confirms the crystalline nature and purity of both CmNPs and SpNPs. FTIR results demonstrate the involvement of the biologically active substances of both plant extracts in the green synthesis of AgNPs. According to MIC and MBC results, higher antimicrobial effects were seen for CmNPs with a smaller size than SpNPs. In addition, CmNPs and SpNPs were much less cytotoxic when examined against a normal cell relative to cAgNPs. Based on high efficacy in controlling antibiotic-resistant pathogens without detrimental adverse effects, CmNPs may have the capacity to be used in medicine as imaging, drug carrier, and antibacterial and anticancer agents.

The photocatalytic, cytotoxicity, and antibacterial properties of zinc oxide nanoparticles synthesized using Trigonella foenum-graecum L extract.

In this study, the manufacture of zinc oxide nanoparticles (ZnO-NPs) was completed via the sol-gel method with Trigonella foenum-graecum L extract for the first time to function as the stabilizing and reducing agent. The obtained product was investigated by various analyzing procedures such as TGA/DTG, FT-IR, UV-Vis, XRD, and EDX/FESEM. The calcination of our product was conducted at temperatures of 400, 500, and 600 °C. In conformity to the XRD pattern, heightening the temperature of calcination caused an enlargement in the size of nanoparticles. The photocatalytic performance of ZnO-NPs was evaluated to degrade methylene blue and Eriochrome black T (EBT) dyes under UV light, which resulted in a degradation percentage of about 96% and 94%, after 90 min, respectively. There has been some evidence suggesting that the green synthesis of ZnO-NPs has increased their use in medicine. The outcomes of examining the cytotoxicity effect of this product against the Huh-7 cell line by the performance of the MTT assay were indicative of an IC50 of around 62.5 µg/mL. Finally, according to the results of the broth microdilution method, which was performed to assess the antibacterial activity of ZnO-NPs towards gram-positive and gram-negative bacteria, the value of MIC was in the range of 31 to 125 µg/mL. The obtained results from biological studies confirm the antibacterial and anticancer properties of ZnO-NPs, which are promising for applying NPs in medical fields.

Comparative evaluation of incorporation calcium silicate and calcium phosphate nanoparticles on biomimetic dentin remineralization and bioactivity in an etch-and-rinse adhesive system.

Background: This study aimed to evaluate the remineralization potential and bioactivity of adhesives, containing amorphous calcium phosphate (ACP) and calcium silicate (CS) nanoparticles (NPs). Material and Methods: In this study, dentin slices (n=60) were prepared and etched with phosphoric acid. Next, they were divided into two groups: pre- and post-immersion in a simulated body fluid (SBF) for three weeks. The two groups were also divided into five subgroups (n=6 per subgroup), including the control (0 wt.% NPs); adhesives containing 1 wt.% and 2.5 wt.% (CS) nanoparticles; and adhesives containing 1 wt.% and 2.5 wt.% ACP nanoparticles. The remineralization potential and bioactivity of the adhesives were evaluated. The shear bond strength of the samples (n=18) was also assessed using a universal testing machine. Results: The present results revealed that the adhesive containing ACP and CS nanoparticles showed bioactivity and remineralization potential without any reduction in the bond strength. Conclusions: The outcomes revealed that Cs and ACP nanoparticles induced mineralization in the dentin and incorporation of these nanoparticles to dentin bonding agents could improve the bio-functionalization of dentin bond. Key words:Calcium phosphate, calcium silicate, fourier transform infrared spectroscopy, scanning electron microscopy, tooth remineralization.

Facile green synthesis of Ag-doped ZnO/CaO nanocomposites with Caccinia macranthera seed extract and assessment of their cytotoxicity, antibacterial, and photocatalytic activity.

The current paper exhibited a green method for the manufacture of Ag-doped ZnO/CaO nanocomposites (NCPs) by the usage of Caccinia macranthera seed extract, zinc, calcium, and silver salts solution, for the first time. The chemical structure of NCPs was studied by the FT-IR technique. The XRD pattern shows a crystallite structure with an Fm3m group space and particle size of about 23 nm. The FESEM/PSA images displayed that NCPs have uniform distribution with spherical morphology. Also, the cytotoxicity of synthesized NCPs was examined on Huh-7 cells by MTT test and the IC50 value was 250 ppm. Additionally, the photocatalytic activity of NCPs was investigated to the methylene blue MB dye degradation, which resulted in a removal of about 90% after 100 min. According to the results of the broth microdilution process, which was done to evaluate the antibacterial activity of NCPs towards gram-positive and gram-negative bacteria, the MIC values were in the range of 0.97-125 ppm.

Effective features extraction by analyzing heart sound for identifying cardiovascular diseases related to COVID-19: A diagnostic model.

Incidence and exacerbation of some of the cardiovascular diseases in the presence of the coronavirus will lead to an increase in the mortality rate among patients. Therefore, early diagnosis of such diseases is critical, especially during the COVID-19 pandemic (mild COVID-19 infection). Thus, for diagnosing the heart diseases related to the COVID-19, an automatic, non-invasive, and inexpensive method based on the heart sound processing approach is proposed. In the present study, a set of features related to the nature of heart signals is defined and extracted. The investigated features included morphological and statistical features in the heart sound frequencies. By extracting and selecting a set of effective features related to the mentioned diseases, and avoiding to use different segmentation and filtering techniques, dependence on a limited dataset and specific sampling procedures has been eliminated. Different classifiers with various kernels are applied for diagnosis in data unbalanced and balanced conditions. The results showed 93.15% accuracy and 93.72% F1-score using 60 effective features in data balanced conditions. The identification system using the extracted features from Azad dataset is able to achieve the desired results in a generalized dataset. In this way, in the shortest possible sampling time, the present system provided an effective and generalizable method and a practical model for diagnosing important cardiovascular diseases in the presence of coronavirus in the COVID-19 pandemic.

Green synthesis of selenium nanoparticles using Rosmarinus officinalis and investigated their antimicrobial activity.

The interest of many has been attracted by plant-mediated synthesizing procedures for nanoparticles since they provide certain qualities including being cost-effective, quick, and compatible with the environment. In this regard, this work introduces the production of selenium-nanoparticles (Se-NPs) in a biological manner utilizing aqueous extracts of Rosmarinus officinalis (R. officinalis). Production of Se-NPs was confirmed using UV-visible (UV-Vis) spectrophotometry. Also, dynamic light scattering (DLS) analysis was used for determination particle size distribution, while we distinguished the identification of crystalline construction of nanoparticles through the means of X-ray diffraction (XRD) pattern, DLS, and transmission electron microscopy (TEM) examination indicated that Se-NPs are often spherical with a size about 20 to 40 nm. The minimum inhibitory concentration (MIC) of the synthesized Se-NPs by R. officinalis extract against Mycobacterium tuberculosis (M. tuberculosis), Staphylococcus aureus (S. aureus), Streptococcus mutans (S. mutans), Escherichia coli (E. coli), and Pseudomonas aeruginosa (P. aeruginosa) was 256, 16, 32, 128, and 64 µg/mL, respectively. The synthesized Se-NPs had no significant effect on Mycobacterium simiae (M. simiae) and had exhibited a strong antimicrobial functionality towards the gram-positive and gram-negative bacteria and can stand as a potent antibacterial agent.

Administration of Silver Nanoparticles in Diabetes Mellitus: A Systematic Review and Meta-analysis on Animal Studies.

Biological features of silver nanoparticles in rising the insulin level of diabetic animal models were considered in recent years, which resulted in decreasing hyperglycemia condition. We reviewed the published literature to investigate the possible role of silver nanoparticles (Ag-NPs) throughout the treatment of diabetes mellitus in animal studies. In this systematic review and meta-analysis, we performed a search throughout the English literature of electronic databases, including Scopus, PubMed, and ISI Web of Science, up to the date of May 22, 2020. Primary outcomes and data regarding fast blood sugar (FBS), lipid profile, and liver enzyme were collected from the available articles, while the studies that did not provide sufficient information on the effects of silver nanoparticles through the course of diabetes mellitus were excluded. Our search yielded 1283 results that included five animal studies in the meta-analysis. The comparison between the plasma insulin level of the diabetic group treated by Ag-NPs with the diabetic control group displayed no significant differences with the P values = 0.299. In addition, significant differences were revealed by comparing the FBS level of the diabetic group treated by Ag-NPs with the diabetic control group (P value < 0.001). According to the present meta-analysis, the application of Ag-NPs in animal models resulted in displaying the anti-diabetic effects, which can be applied in future treatments. Furthermore, a correlation was noticed between these nanoparticles and the reduction of serum FBS among diabetic cases.

Green synthesis of nickel oxide nanoparticles using Salvia hispanica L. (chia) seeds extract and studies of their photocatalytic activity and cytotoxicity effects.

The physical and chemical properties of Nickel oxide nanoparticles (NiO-NPs) have attracted the attention of many and in this regard, this study was performed to produce NiO-NPs by the means of Salvia hispanica L. (chia) seeds extract as the capping agent. Physical and morphological features of the obtained NiO-NPs were examined through the application of TGA, FTIR, UV-Vis, XRD, FESEM/EDAX/PSA, and VSM procedures. According to the FESEM/PSA images, the biosynthesized NiO-NPs contained a spherical shape and a size of about 30 nm, while the results of the EDAX study approved the existence of oxygen and nickel elements in the structure of this product. Furthermore, certain corresponding peaks to the crystal structure of NiO-NPs were observed throughout the XRD pattern. Next to the superparamagnetic behavior that was detected in the results of VSM analysis, the cytotoxicity effect of NiO-NPs was not reported to be dependent on concentration. Considering the high photocatalytic capacity along with the low cytotoxic effects of NiO-NPs, we can suggest the applicability of this product for various applications such as disease control and removal of residual toxins.

Green synthesis of colloidal selenium nanoparticles in starch solutions and investigation of their photocatalytic, antimicrobial, and cytotoxicity effects.

In this research, we have offered a green and new synthesizing procedure for selenium nanoparticles (Se-NPs) through the utilization of Na2SeO3, in which starch has a role of stabilizer and capping agent, as the functionality of green reducing mediums is taken by glucose and ascorbic acid. According to the observations, this method has been capable of producing Se-NPs in lab conditions. Additionally, the synthesized Se-NPs can be separated from the aqueous solution of stabilizer and reducing agents by a high-speed. Certain analyzing procedures have been used to characterize the obtained particles including TEM, XRD, UV-VIS, DLS, FESEM, EDX, FTIR, and AFM. In this paper, we have investigated the antimicrobial and photocatalytic functionality of Se-NPs on Mycobacterium tuberculosis and Methylene blue (MB) and according to the results, these particles have shown satisfying activity in both cases. To be stated in exact, about 60% of MB has degraded under UV light after 150 min, which indicates the acceptable position of Se-NPs could be applied for eliminating water pollutions. Moreover, the attained data on colorectal cancer SW480 cell lines in regards to the in vitro cytotoxicity assessments have exhibited non-toxic effects, which had lasted throughout concentrations that had measured up to even 100 µg/mL within MTT assay.

Tragacanth-mediate synthesis of NiO nanosheets for cytotoxicity and photocatalytic degradation of organic dyes.

In this study, NiO nanosheets have been manufactured using a co-precipitation approach that involved the usage of nickel nitrate (Ni (NO3)2.6H2O) as the raw material and tragacanth in the role of a stabilizing agent. NiO nanosheets have been fabricated through the reduction of nickel nitrate solution that had been obtained by the application of aqueous extract of tragacanth, which is capable of functioning as a reducing and stabilizing agent. In the following, the physical and chemical properties of tragacanth-stabilized NiO nanosheets have been identified via FESEM, EDS, XRD, UV-Vis, and FT-IR techniques. According to the XRD pattern, these particular nanosheets have contained a cubic structure and group space Fm3m, along with the average size of about 18 to 43 nm that had been in agreement with the FESEM measurements. In addition, we have evaluated the photocatalytic activity of tragacanth-stabilized NiO nanosheets on the degradations of methylene blue (MB) and methyl orange (MO) dyes. The performed photocatalytic assessment has displayed that the nanosheets can degrade 82% of MO within 210 min and 60% of MB in 300 min. The cytotoxicity of tragacanth-stabilized NiO nanosheets on human Glioblastoma cancer (U87MG) cell lines has been investigated via the MTT assay, while it has been detected in the obtained results that the inhibitory concentration (IC50) had been 125 µg/mL.

IgD attenuates the IgM-induced anergy response in transitional and mature B cells.

Self-tolerance by clonal anergy of B cells is marked by an increase in IgD and decrease in IgM antigen receptor surface expression, yet the function of IgD on anergic cells is obscure. Here we define the RNA landscape of the in vivo anergy response, comprising 220 induced sequences including a core set of 97. Failure to co-express IgD with IgM decreases overall expression of receptors for self-antigen, but paradoxically increases the core anergy response, exemplified by increased Sdc1 encoding the cell surface marker syndecan-1. IgD expressed on its own is nevertheless competent to induce calcium signalling and the core anergy mRNA response. Syndecan-1 induction correlates with reduction of surface IgM and is exaggerated without surface IgD in many transitional and mature B cells. These results show that IgD attenuates the response to self-antigen in anergic cells and promotes their accumulation. In this way, IgD minimizes tolerance-induced holes in the pre-immune antibody repertoire.

Redemption of autoantibodies on anergic B cells by variable-region glycosylation and mutation away from self-reactivity.

The best-understood mechanisms for achieving antibody self/non-self discrimination discard self-reactive antibodies before they can be tested for binding microbial antigens, potentially creating holes in the repertoire. Here we provide evidence for a complementary mechanism: retaining autoantibodies in the repertoire displayed as low levels of IgM and high IgD on anergic B cells, masking a varying proportion of autoantibody-binding sites with carbohydrates, and removing their self-reactivity by somatic hypermutation and selection in germinal centers (GCs). Analysis of human antibody sequences by deep sequencing of isotype-switched memory B cells or in IgG antibodies elicited against allogeneic RhD+ erythrocytes, vaccinia virus, rotavirus, or tetanus toxoid provides evidence for reactivation of anergic IgM(low) IgD+ IGHV4-34+ B cells and removal of cold agglutinin self-reactivity by hypermutation, often accompanied by mutations that inactivated an N-linked glycosylation sequon in complementarity-determining region 2 (CDR2). In a Hy10 antibody transgenic model where anergic B cells respond to a biophysically defined lysozyme epitope displayed on both foreign and self-antigens, cell transfers revealed that anergic IgM(low) IgD+ B cells form twice as many GC progeny as naïve IgM(hi) IgD+ counterparts. Their GC progeny were rapidly selected for CDR2 mutations that blocked 72% of antigen-binding sites with N-linked glycan, decreased affinity 100-fold, and then cleared the binding sites of blocking glycan. These results provide evidence for a mechanism to acquire self/non-self discrimination by somatic mutation away from self-reactivity, and reveal how varying the efficiency of N-glycosylation provides a mechanism to modulate antibody avidity.

Decrease in topoisomerase I is responsible for activation-induced cytidine deaminase (AID)-dependent somatic hypermutation.

Somatic hypermutation (SHM) and class-switch recombination (CSR) of the Ig gene require both the transcription of the locus and the expression of activation-induced cytidine deaminase (AID). During CSR, AID decreases the amount of topoisomerase I (Top1); this decrease alters the DNA structure and induces cleavage in the S region. Similarly, Top1 is involved in transcription-associated mutation at dinucleotide repeats in yeast and in triplet-repeat contraction in mammals. Here, we report that the AID-induced decrease in Top1 is critical for SHM. Top1 knockdown or haploinsufficiency enhanced SHM, whereas Top1 overexpression down-regulated it. A specific Top1 inhibitor, camptothecin, suppressed SHM, indicating that Top1's activity is required for DNA cleavage. Nonetheless, suppression of transcription abolished SHM, even in cells with Top1 knockdown, suggesting that transcription is critical. These results are consistent with a model proposed for CSR and triplet instability, in which transcription-induced non-B structure formation is enhanced by Top1 reduction and provides the target for irreversible cleavage by Top1. We speculate that the mechanism for transcription-coupled genome instability was adopted to generate immune diversity when AID evolved.