Clean synthesis of silver nanoparticles (AgNPs) on polyamide fabrics by Verbascum thapsus L. (mullein) extract: characterization, colorimetric, antibacterial, and colorfastness studies.

The production of antibacterial colored textiles using nanomaterials (NMs) has become an ideal goal from both a research and industrial perspective. In this study, the clean synthesis and characterization of silver nanoparticles (AgNPs) on polyamide fabrics were performed using mullein extract for the first time. Natural dyes were extracted from mullein leaves using an ultrasonic method, with an optimal amount of 15 g/L. The synthesized AgNPs in different ratios of mullein extract and Ag ions were analyzed (using UV-visible spectroscopy) and dynamic light scattering (DLS). It was found that AgNPs synthesized with a ratio of 1:4 of mullein extract: to Ag ions had a diameter of 85 nm. The active site groups of the synthesized AgNPs were characterized using Fourier transform infrared spectroscopy (FT-IR). Nylon fabrics dyed with different ratios of mullein extract and Ag ions exhibited acceptable color strength values (K/S) of 3.36. Furthermore, the reduction in bacterial growth for dyed fabrics improved with an increase in the ratio of Ag ions, with a 100% reduction observed for a sample dyed with mullein extract: Ag ions at a ratio of 1:4. Overall, this method offers a simple, low-cost, and compatible process with environment without the consumption of any chemicals to producing nylon with acceptable antibacterial and dyeing properties.

Wool dyeing using Ziziphus bark extract as a natural dye: studies on the dyeing, antibacterial, and antioxidant characteristics.

Considering the growing importance of natural colorants and sustainable products, the research on application of natural dyes has been focused on new color resources, identification, and standardization of natural dyes. Hence, the extraction of natural colorants available in Ziziphus bark was performed by ultrasound technique and its application on the wool yarn to produce the antioxidant and antibacterial fibers. The optimal conditions for the extraction process were as follows: ethanol/water (1/2 v/v) as solvent, concentration of Ziziphus dye 14 g/L, pH 9, temperature 50 °C, time 30 min, and L.R ratio 50:1. Moreover, the effect of important variables for application of Ziziphus dye on the wool yarn was investigated and optimized temperature 100 °C, concentration of Ziziphus dye 50% o.w.f., time for dyeing 60 min and pH 8, and L.R 30:1. The reduction values of Gram-negative and Gram-positive bacteria on dyed samples at optimized condition were 85% and 76%, respectively. Moreover, the antioxidant property of dyed sample was 78%. The color variations on the wool yarn were produced with diverse metal mordants, and color fastness properties were measured. Ziziphus dye not only can be used as an origin for a natural dye, but also provided the antibacterial and antioxidant agent on the wool yarn, which can be a step towards the fabrication of green products.

Using a Dendritic Sensor as a Feasible Method for Detection of Copper in Water Samples.

Due to the severe dangers of copper on human health, development of identification and measurement copper methods in aquatic environments as well as prevention transfer methods of these substances to water resources have received much attention. Among of this expansion, rapid, simple, safe, sensitive and economically viable detection methods are more considerable. In this paper, the sensor activity of the poly(amidoamine) dendrimer containing eight methoxy groups modified with naphthalimide fluorescent derivative is studied. The sensor potential of dendrimer-naphthalimide fluorescence was evaluated in various solvents with different polarities, and results indicated that the detection of cuprum cations (Cu2+) is possible at low concentration. Also, the fluorescence properties of dendrimer-naphthalimide system are measured in the presence of diverse metal cations. In light of above, from the obtaining results, it can be concluded that dendritic sensor can be used as a feasible method to detect of copper in water samples in low concentrations.

Isolation of Innate Lymphoid Cells from Murine Intestinal Lamina Propria.

Natural killer (NK) cells are innate cytotoxic immune cells essential for mediating first-line defense against various environmental antigens. With the discoveries of other subsets of innate lymphocytes over the last decade, NK cells are categorized as innate lymphoid cells (ILC) and as the innate counterparts of cytotoxic T cells. Besides NK cells, ILCs are classified into three groups distinguished by their dependence on distinct transcription factors for development and unique effector functions. Subsets of ILCs share many surface proteins that, however, have initially been identified as NK cell markers, making them hard to be distinguished for detailed investigations. Here, we describe a method to identify and individually isolate subsets of innate lymphoid cells from gut lamina propria using cell surface markers.

Methods to Analyze the Developmental Trajectory of Human Primary NK Cells Using Monocle and SCENIC Analyses.

Development of novel cellular therapies based on primary human NK cells is under active investigation. Human NK cells are comprised of distinct subsets with high transcriptomic heterogeneity. Unique methodologies are being developed to determine the transcriptomic profiles of human NK cells. NK cells account for 10-20% of total lymphocytes in the human peripheral blood, which mediates anti-tumor and anti-viral effector functions. Therapeutic success in the clinic depends on a better understanding of the single-cell transcriptome of human NK cell subsets. Moreover, a better understanding of the transcriptional network that regulates NK cell development, subset specification, and terminal maturation is obligatory for their in vitro generation and expansion toward clinical utilization. Here, we describe the procedure for single-cell RNA-sequencing of human NK cells and strategies for bioinformatic analyses. This protocol provides a data analysis roadmap for investigators who work on the basic biology and therapeutic applications of human NK cells.

Methods for Isolating and Defining Single-Cell Transcriptomes of Tissue-Resident Human NK Cells.

Natural killer (NK) cells are innate lymphocytes that control tumors and microbial infections. Human NK cells are transcriptomically and phenotypically heterogeneous. The site where NK cells develop and reside determines their phenotype and effector functions. Our current knowledge about human NK cells is primarily from blood- and bone marrow-derived NK cells. The major limitation in formulating organ-specific clinical therapy is the knowledge gap on how tissue-resident NK cells develop, home, and function. Thus, it is crucial to define the transcriptomic profiles and the transcriptional regulation of tissue-resident NK cells. The major challenges in studying tissue-resident NK cells include their total number and the complexity of the tissue. Additionally, during isolation, keeping them viable and naïve without activation are challenging tasks. Here, we provide methods for isolating and performing transcriptomic analyses of NK cells at the individual cell level. Single-cell RNA sequencing provides a higher resolution of cellular heterogeneity and a better understanding of cell-cell interactions within the microenvironment. Using these methods, we can efficiently identify distinct populations of NK cells in tissues and define their unique transcriptomic profiles.

Method to Study Adaptive NK Cells Following MCMV Infections.

Immunological memory is a fundamental feature of the adaptive immune system that protects the host from recurrent infections from pathogens. Natural killer (NK) cells are a predominant member of the innate immune system that lack clonotypic receptors, which are essential for memory formation. However, evidence demonstrates that a unique subpopulation of NK cells develops adaptive-like features using germline-encoded receptors. Recent studies have shown that infection of cytomegalovirus (CMV) leads to clonal expansion of NKG2C+ and Ly49H+ NK cells, in humans and mouse, respectively. These activation receptors have the capability to recognize CMV-encoded proteins and facilitate a recall response upon reinfection. Although NK cells do not rearrange genes encoding their activating receptors as seen in B and T cells, they possess a selective process to generate memory features and a long-lived progeny. Here, we describe an established in vivo protocol for infecting mice with mouse cytomegalovirus (MCMV) to study an adaptive NK cell response.

Application of Achillea millefolium extract as a reducing agent for synthesis of silver nanoparticles (AgNPs) on the cotton: antibacterial, antioxidant and dyeing studies.

The phyto-synthesis of silver nanoparticles and cotton dyeing with natural colorants can reduce the environmental impact of the process considerably. In this study, the extraction of natural colorants from Achillea millefolium petals was optimized by ultrasound technique. The AMP extract was applied for synthesis of silver nanoparticles (Ag NPs) on the cotton fabrics. The dyeing, antibacterial and antioxidant characteristics of cotton samples were investigated to optimize the process and evaluate its efficiency. The AMP extract had good substantivity towards cotton fabrics and the presence of tannic acid, as an environmentally-friendly mordant, further improved the absorption of AMP dye. The antibacterial and antioxidant activities of the dyed samples with AMP extract of were 50%and 60%, respectively. The addition of TA and Ag enhanced the antibacterial and antioxidant activities on the cotton samples to over 99%.

Implications of a 'Third Signal' in NK Cells.

Innate and adaptive immune systems are evolutionarily divergent. Primary signaling in T and B cells depends on somatically rearranged clonotypic receptors. In contrast, NK cells use germline-encoded non-clonotypic receptors such as NCRs, NKG2D, and Ly49H. Proliferation and effector functions of T and B cells are dictated by unique peptide epitopes presented on MHC or soluble humoral antigens. However, in NK cells, the primary signals are mediated by self or viral proteins. Secondary signaling mediated by various cytokines is involved in metabolic reprogramming, proliferation, terminal maturation, or memory formation in both innate and adaptive lymphocytes. The family of common gamma (γc) cytokine receptors, including IL-2Rα/ß/γ, IL-7Rα/γ, IL-15Rα/ß/γ, and IL-21Rα/γ are the prime examples of these secondary signals. A distinct set of cytokine receptors mediate a 'third' set of signaling. These include IL-12Rß1/ß2, IL-18Rα/ß, IL-23R, IL-27R (WSX-1/gp130), IL-35R (IL-12Rß2/gp130), and IL-39R (IL-23Rα/gp130) that can prime, activate, and mediate effector functions in lymphocytes. The existence of the 'third' signal is known in both innate and adaptive lymphocytes. However, the necessity, context, and functional relevance of this 'third signal' in NK cells are elusive. Here, we define the current paradigm of the 'third' signal in NK cells and enumerate its clinical implications.

Role of GATA2 in Human NK Cell Development.

Natural killer (NK) cells are major innate lymphocytes. NK cells do not require prior antigen exposure to mediate antitumor cytotoxicity or proinflammatory cytokine production. Since they use only nonclonotypic receptors, they possess high clinical value in treatment against a broad spectrum of malignancies. Irrespective of this potential, however, the transcriptional regulation that governs human NK cell development remains far from fully defined. Various environmental cues initiate a complex network of transcription factors (TFs) during their early development, one of which is GATA2, a master regulator that drives the commitment of common lymphoid progenitors (CLPs) into immature NK progenitors (NKPs). GATA2 forms a core heptad complex with six other TFs (TAL1, FLI1, RUNX1, LYL1, LMO2, and ERG) to mediate its transcriptional regulation in various cell types. Patients with GATA2 haploinsufficiency specifically lose CD56bright NK cells, with or without a reduced number of CD56dlm NK cells. Here, we review the recent progress in understanding GATA2 and its role in human NK cell development and functions.

NK Cell Development and Function in Patients with Fanconi Anemia.

Fanconi anemia (FA) is an inherited disorder characterized by diverse congenital malformations, progressive pancytopenia, and predisposition to hematological malignancies and solid tumors. The role of the Fanconi anemia pathway in DNA repair mechanisms and genome instability is well studied. However, the consequences of inherited mutations in genes encoding the FA proteins and the acquired mutations due to impaired DNA repair complex in immune cells are far from understood. Patients with FA show bone marrow failure (BMF) and have a higher risk of developing myelodysplasia (MDS) or acute myeloid leukemia (AML) which are directly related to having chromosomal instability in hematopoietic stem cells and their subsequent progeny. However, immune dysregulation can also be seen in FA. As mature descendants of the common lymphoid progenitor line, NK cells taken from FA patients are dysfunctional in both NK cell-mediated cytotoxicity and cytokine production. The molecular bases for these defects are yet to be determined. However, recent studies have provided directions to define the cause and effect of inherited and acquired mutations in FA patients. Here, we summarize the recent studies in the hematopoietic dysfunction, focusing on the impairment in the development and functions of NK cells in FA patients, and discuss the possible mechanisms and future directions.

Eco-Friendly Procedure for Rendering the Antibacterial and Antioxidant of Cotton Fabrics via Phyto-Synthesized AgNPs With Malva sylvestris (MS) Natural Colorant.

There is a growing interest for producing multifunctional cellulose fabrics using green and sustainable technology. In this study, we explored an eco-friendly procedure for dyeing cotton fabrics with Malva sylvestris (MS) as a natural colorant and rendering antibacterial cotton fabric by the silver nanoparticles. MS colorants were extracted from dried petals in water using the ultrasound technique, cotton fabrics were dyed with the extracted MS colorant at 100°C for 90 min. The colorimetric data and colorfastness properties were investigated in the absence and presence of tannic acid (TA) as a bio-mordant. Results indicated that MS dye had a high potential for reducing the silver nitrate, so that the silver particle size distribution on cotton fabric was obtained 50-80 nm, and TA had a positive effect on the MS extract and reduced Ag on the cotton. Furthermore, the reduction of bacterial growth of the dyed cotton considerably (up to 99%) improved by AgNPs. The wash-, and light-fastness properties of samples dyed with MS were enhanced from moderate to good-very good by mordanting.

Conditional Deletion of PGC-1α Results in Energetic and Functional Defects in NK Cells.

During an immune response, natural killer (NK) cells activate specific metabolic pathways to meet the increased energetic and biosynthetic demands associated with effector functions. Here, we found in vivo activation of NK cells during Listeria monocytogenes infection-augmented transcription of genes encoding mitochondria-associated proteins in a manner dependent on the transcriptional coactivator PGC-1α. Using an Ncr1Cre-based conditional knockout mouse, we found that PGC-1α was crucial for optimal NK cell effector functions and bioenergetics, as the deletion of PGC-1α was associated with decreased cytotoxic potential and cytokine production along with altered ADP/ATP ratios. Lack of PGC-1α also significantly impaired the ability of NK cells to control B16F10 tumor growth in vivo, and subsequent gene expression analysis showed that PGC-1α mediates transcription required to maintain mitochondrial activity within the tumor microenvironment. Together, these data suggest that PGC-1α-dependent transcription of specific target genes is required for optimal NK cell function during the response to infection or tumor growth.

Tissue-Resident NK Cells: Development, Maturation, and Clinical Relevance.

Natural killer (NK) cells belong to type 1 innate lymphoid cells (ILC1) and are essential in killing infected or transformed cells. NK cells mediate their effector functions using non-clonotypic germ-line-encoded activation receptors. The utilization of non-polymorphic and conserved activating receptors promoted the conceptual dogma that NK cells are homogeneous with limited but focused immune functions. However, emerging studies reveal that NK cells are highly heterogeneous with divergent immune functions. A distinct combination of several activation and inhibitory receptors form a diverse array of NK cell subsets in both humans and mice. Importantly, one of the central factors that determine NK cell heterogeneity and their divergent functions is their tissue residency. Decades of studies provided strong support that NK cells develop in the bone marrow. However, evolving evidence supports the notion that NK cells also develop and differentiate in tissues. Here, we summarize the molecular basis, phenotypic signatures, and functions of tissue-resident NK cells and compare them with conventional NK cells.

Designing a new biosensor "DNA ELISA" to detect Escherichia coli using genomic DNA and comparison of this method to PCR-ELISA.

In this experiment, DNA-ELISA biosensor was introduced, bearing the ability to detect specific bacteria in about 4 h. This is a more rapid system in comparison to conventional methods, like colony counting method. Moreover, this method does not require any amplification and directly detects genomic DNA of bacteria, giving a lower limit to the sensitivity of 40,000 bacteria. In this study, two specific probes capture (biotin labelled) and detector (dig labelled), were used against special regions of 16s rRNA gene of Escherichia coli ATCC 25922. The capture probe has the ability to trap the target bacterial DNA from a pool of other kinds of bacteria under specific conditions. The detector probe then was used to hybridize to the genome of trapped bacteria. The detection proceeds by adding HRP-anti dig enzyme and its substrate, ABTS to emit light. Light absorbance is measured for verifying the detection.

Negishi coupling: an easy progress for C-C bond construction in total synthesis.

Negishi cross-coupling reactions were used extensively in total synthesis. The expansion of substrate scope, the development of mild reaction conditions, the advancement of the methods to improve the stereo- and regio-selectivity of carbon-carbon bond formation, the maturity of a large number of sequential processes, and the development of non-toxic reactions signify the importance of Negishi coupling. The following review illustrates a strategic role of this reaction in constructing carbon-carbon bonds in the recent total synthesis of natural products.