Fluorescent labeling of micro/nanoplastics for biological applications with a focus on "true-to-life" tracking.

The increased environmental presence of micro-/nanoplastics (MNPLs) and the potential health risks associated with their exposure classify them as environmental pollutants with special environmental and health concerns. Consequently, there is an urgent need to investigate the potential risks associated with secondary MNPLs. In this context, using "true-to-life" MNPLs, resulting from the laboratory degradation of plastic goods, may be a sound approach. These non-commercial secondary MNPLs must be labeled to track their presence/journeys inside cells or organisms. Because the cell internalization of MNPLs is commonly analyzed using fluorescence techniques, the use of fluorescent dyes may be a sound method to label them. Five different compounds comprising two chemical dyes (Nile Red and Rhodamine-B), one optical brightener (Opticol), and two industrial dyes (Amarillo Luminoso and iDye PolyPink) were tested to determine their potential for such applications. Using commercial standards of polystyrene nanoplastics (PSNPLs) with an average size of 170 nm, different characteristics of the selected dyes such as the absence of impact on cell viability, specificity for plastic staining, no leaching, and lack of interference with other fluorochromes were analyzed. Based on the overall data obtained in the wide battery of assays performed, iDye PolyPink exhibited the most advantages, with respect to the other compounds, and was selected to effectively label "true-to-life" MNPLs. These advantages were confirmed using a proposed protocol, and labeling titanium-doped PETNPLs (obtained from the degradation of milk PET plastic bottles), as an example of "true-to-life" secondary NPLs. These results confirmed the usefulness of iDye PolyPink for labeling MNPLs and detecting cell internalization.

Altered desensitization and internalization patterns of rodent versus human glucose-dependent insulinotropic polypeptide (GIP) receptors. An important drug discovery challenge.

BACKGROUND AND PURPOSE: The gut hormone glucose-dependent insulinotropic polypeptide (GIP) signals via the GIP receptor (GIPR), resulting in postprandial potentiation of glucose-stimulated insulin secretion. The translation of results from rodent studies to human studies has been challenged by the unexpected effects of GIPR-targeting compounds. We, therefore, investigated the variation between species, focusing on GIPR desensitization and the role of the receptor C-terminus. EXPERIMENTAL APPROACH: The GIPR from humans, mice, rats, pigs, dogs and cats was studied in vitro for cognate ligand affinity, G protein activation (cAMP accumulation), recruitment of beta-arrestin and internalization. Variants of the mouse, rat and human GIPRs with swapped C-terminal tails were studied in parallel. KEY RESULTS: The human GIPR is more prone to internalization than rodent GIPRs. Despite similar agonist affinities and potencies for Gαs activation, especially, the mouse GIPR shows reduced receptor desensitization, internalization and beta-arrestin recruitment. Using an enzyme-stabilized, long-acting GIP analogue, the species differences were even more pronounced. 'Tail-swapped' human, rat and mouse GIPRs were all fully functional in their Gαs coupling, and the mouse GIPR regained internalization and beta-arrestin 2 recruitment properties with the human tail. The human GIPR lost the ability to recruit beta-arrestin 2 when its own C-terminus was replaced by the rat or mouse tail. CONCLUSIONS AND IMPLICATIONS: Desensitization of the human GIPR is dependent on the C-terminal tail. The species-dependent functionality of the C-terminal tail and the different species-dependent internalization patterns, especially between human and mouse GIPRs, are important factors influencing the preclinical evaluation of GIPR-targeting therapeutic compounds.

Weight bias among Canadians: Associations with sociodemographics, BMI and body image constructs.

This cross-sectional study examined the associations between sociodemographic characteristics, BMI, and body image constructs (body satisfaction and weight bias internalization; WBI) and explicit weight bias. A near-representative sample of 995 English-speaking Canadian adults (52% Female) completed a survey which assessed explicit weight bias (Anti-Fat Attitudes questionnaire), body satisfaction (Body Shape Satisfaction Scale), WBI (Modified Weight Bias Internalization Scale), and self-reported height and weight. Multiple linear regression analyses were run. Results showed that the variable that explained the most variance in explicit weight bias was WBI, followed by BMI. Higher levels of WBI and a lower BMI were both significantly associated with greater explicit weight bias. Male sex was associated with both disliking people with obesity and thinking obesity is attributable to lack of willpower, whereas female sex was associated with worrying about weight gain. The current findings emphasize the importance of future research efforts aimed at preventing or mitigating WBI to reduce negative attitudes about people with obesity.

Xiaowugui decoction alleviates experimental rheumatoid arthritis by suppressing Rab5a-mediated TLR4 internalization in macrophages.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by exacerbated synovial inflammation and joint destruction. Recent studies suggest toll-like receptor 4 (TLR4) internalization facilitate inflammatory response of macrophage. The role of TLR4 internalization in the pathogenesis of RA is unknown. PURPOSE: To investigate the role and mechanism of TLR4 internalization in macrophage inflammatory response of RA and explore whether TLR4 internalization mediates the anti-arthritic effect of Xiaowugui (XWG) decoction, a patented herbal formula used in China. METHODS: The co-expression of TLR4 and the internalization marker, early endosome antigen 1 (EEA1), in the synovial samples of RA patients and joint tissue of collagen-induced arthritis (CIA) mice, were evaluated using immunofluorescence. The effect of Rab5a-mediated early internalization of TLR4 on the activation induced by lipopolysaccharide (LPS) in RAW264.7 cells was investigated using small interfering RNAs that act against Rab5a. CIA was induced in Rab5a-/- mice to evaluate the role of Rab5a in vivo. The disease progression and expression of Rab5a and TLR4 in the joint tissue were evaluated in CIA mice treated with XWG. Inflammatory factors production, TLR4 internalization, and activation of downstream signaling pathways were examined in RAW264.7 cells treated with XWG in vitro. RESULTS: The co-expression and co-localization of TLR4 and EEA1 were elevated in the synovial samples of RA patients and joint tissue of CIA mice. Pharmaceutical inhibition of TLR4 internalization reduced macrophages inflammatory responses induced by LPS. The co-expression and co-localization of Rab5a and TLR4 were significantly increased in macrophages treated with LPS. Silencing Rab5a reduced LPS-induced TLR4 internalization, inflammatory factors production, and phosphorylation of Jun N-terminal kinases (JNK) and p65. Genetic deletion of Rab5a inhibited TLR4 internalization and the development of arthritis in vivo. The co-expression of TLR4 and Rab5a was also elevated in the synovial samples of RA patients. XWG treatment of mice with CIA alleviated arthritis and reduced the co-expression of Rab5a and TLR4 in the joint tissue. XWG treatment of macrophage inhibited LPS-induced IL-6 and TNF-α production, co-expression of Rab5a and TLR4, and phosphorylation of JNK and p65. CONCLUSIONS: Our findings highlight the pathogenic role of TLR4 internalization in patients with RA and identify a novel Rab5a-dependent internalization pathway that promotes macrophage inflammatory response. XWG treatment demonstrated outstanding therapeutic effects in experimental arthritis, and targeting the Rab5a-mediated internalization of TLR4 may be the main underlying mechanism.

Experienced and internalized weight stigma among Spanish adolescents.

BACKGROUND AND OBJECTIVE: Weight stigma has negative consequences for both physiological and psychological health. Studies on weight stigma in adolescence, particularly from general populations, are scarce in the Mediterranean area. The main aim of this study is to describe the prevalence of experienced and internalized weight stigma among a representative sample of adolescents from the Spanish city of Terrassa, and to determine its association with sociodemographic variables and weight status. METHODS: Drawing on data from the initial assessment of a longitudinally funded project on weight stigma in adolescents, a cross-sectional survey-based study was conducted using random multistage cluster sampling. Weight stigma experiences, their frequency and sources, and weight bias internalization with the Modified Weight Bias Internalization Scale (WBISM) were assessed in a sample of 1016 adolescents. Adjusted odds ratios (AOR) between sociodemographic variables, weight status and having experienced weight stigma, and having reported high scores of WBISM (WBISM ≥ 4) were estimated by multiple logistic regression models. RESULTS: The prevalence of weight-related stigma experiences was 43.2% in the sample (81.8 in adolescents with obesity) and the prevalence of high levels of weight bias internalization was 19.4% (50.7 in adolescents with obesity). Other kids and school were the most prevalent sources of weight stigma, with society and family being other significant sources of stigma reported by girls. A significantly higher risk of having experienced weight stigma was observed in girls (AOR = 2.6) and in older adolescents (AOR = 1.9). Compared to normal weight adolescents, all weight statuses showed higher risk, being 3.4 times higher in adolescents with underweight and reaching 11.4 times higher risk in those with obesity. Regarding high levels of weight bias internalization, girls had a risk 6.6 times higher than boys. Once again, a "J-shaped" pattern was observed, with a higher risk at the lowest and highest weight statuses. The risk was 6.3 times higher in adolescents with underweight, and 13.1 times higher in adolescents with obesity compared to those with normal weight. CONCLUSIONS: Considering the high prevalence of experienced and internalized weight stigma among adolescents in Spain, especially in adolescents with obesity and girls, it seems important to implement preventive strategies in different settings and address all sources of stigma.

Dissonance-Based Eating Disorder Prevention for Body-Dissatisfied Brazilian Cisgender Gay and Bisexual Men: A Randomized Controlled Trial With a 1-Year Follow-Up.

OBJECTIVE: Gay and bisexual men are at an increased risk for eating disorders (EDs) and muscle dysmorphia (MD) compared with their heterosexual counterparts. Existing dissonance-based (DB) EDs prevention programs for this population have been evaluated in the United States; however, these programs have not been evaluated in the Brazilian context. Thus, we investigated the feasibility, acceptability, and efficacy of a DB ED prevention program (i.e., the PRIDE Body Project) among Brazilian cisgender gay and bisexual men. METHOD: Eligible men were randomly assigned to either a DB intervention (n = 74) condition or an assessment-only control (AOC) condition (n = 75). Participants completed measures assessing ED and MD risk and protective factors at baseline, post-intervention, 1-month, 6-month, and 1-year follow-up. Those in the intervention condition also completed acceptability measures. RESULTS: Feasibility and acceptability ratings were highly favorable. Regarding efficacy, post-intervention results were not significant, except for self-objectification, which showed a significantly greater decrease in the DB condition compared with the AOC condition at all time-points of follow-ups (Cohen's d = -0.31 to -0.76). At follow-up, the DB condition showed significantly greater decreases in appearance-ideal internalization, drive for muscularity, self-objectification, ED and MD symptoms at 1-month, 6-month, and 1-year follow-ups (d = -0.33 to -0.92) compared with the AOC condition. Significant increases were observed in the DB compared with the AOC condition for body appreciation at 1-month, 6-month, and 1-year follow-ups (d = 0.31-0.81). DISCUSSION: Results support the feasibility, acceptability, and efficacy of the PRIDE Body Project up to 1-year in Brazilian cisgender gay and bisexual men. TRIAL REGISTRATION: Brazilian Registry of Clinical Trials (ReBEC; available at http://www.ensaiosclinicos.gov.br/) number of registration: RBR-62fctqz.

A live cell imaging-based assay for tracking particle uptake by clathrin-mediated endocytosis.

A popular strategy for therapeutic delivery to cells and tissues is to encapsulate therapeutics inside particles that cells internalize via endocytosis. The efficacy of particle uptake by endocytosis is often studied in bulk using flow cytometry and Western blot analysis and confirmed using confocal microscopy. However, these techniques do not reveal the detailed dynamics of particle internalization and how the inherent heterogeneity of many types of particles may impact their endocytic uptake. Toward addressing these gaps, here we present a live-cell imaging-based method that utilizes total internal reflection fluorescence microscopy to track the uptake of a large ensemble of individual particles in parallel, as they interact with the cellular endocytic machinery. To analyze the resulting data, we employ an open-source tracking algorithm in combination with custom data filters. This analysis reveals the dynamic interactions between particles and endocytic structures, which determine the probability of particle uptake. In particular, our approach can be used to examine how variations in the physical properties of particles (size, targeting, rigidity), as well as heterogeneity within the particle population, impact endocytic uptake. These data impact the design of particles toward more selective and efficient delivery of therapeutics to cells.

α-Linolenic Acid Vesicles-Mediated Tau Internalization in Microglia.

In Alzheimer's disease, the synaptic loss is prominent due to the accumulation of Amyloid ßeta (Aß) protein in synapses, which affect neurotransmission, and thus ultimately causes neuronal loss. Tau, a microtubule-associated protein, is a vital protein of intracellular neurofibrillary tangles (NFTs) in AD. Along with the accumulation of aberrant proteins, glial cells, mainly astrocytes and microglia, play a major role in impairing neuronal network. Microglia have the ability to phagocytose Tau and rerelease in exosomes, which causes further spreading of Tau. Reduction in exosome synthesis can reduce spreading of Tau. Modulating microglia to clear the extracellular Tau seeds by its imported degradation would resolve the disease condition in Alzheimer's disease. In this study, we have shown the ability of α-linolenic acid (ALA) to inhibit the Tau aggregation and modulate their internalization property in microglial cells.

Cell-penetrating activity of a short-chain ε-poly-l-α-lysine.

Bacteria produce polycationic homopoly(amino acid)s, which are characterized by isopeptide backbones. We previously demonstrated that two representative bacterial polycationic isopeptides, ε-poly-l-α-lysine consisting of 25-35 l-α-lysine residues (ε-PαL25-35) and ε-poly-l-ß-lysine consisting of l-ß-lysine residues (ε-PßL4-13), were internalized into mammalian cells by both energy-independent direct penetration and energy-dependent endocytosis/macropinocytosis, and then diffused throughout the cytosol. In this study, we investigated the cell-penetrating activity of an ε-PαL short-chain derivative consisting of 5-14 l-α-lysine residues (ε-PαL5-14) to gain insight into the relationship between the isopeptide-chain length and the manner of cellular internalization. We prepared a conjugate of ε-PαL5-14 and a fluorescent dye (FAM) by click chemistry, and incubated the resulting polymer, ε-PαL5-14-FAM, with HeLa cells. Unlike ε-PαL25-35-FAM, ε-PαL5-14-FAM was internalized into cells only by energy-dependent endocytosis/macropinocytosis. Furthermore, a high concentration (>50 µM) was required for the internalization events. ε-PαL5-14 has a chain length almost equal to that of the membrane permeable ε-PßL4-13, which can enter cells at low concentrations. Considering that the basicity of the ß-amino group is higher than that of α-amino acid at physiological pH, ε-PßL is expected to have a greater cell-penetrating capacity than ε-PαL, provided their isopeptide-chain lengths are similar, suggesting that a more extended chain derivative of ε-PßL would be more advantageous for cellular internalization of cargo proteins than ε-PαL25-35.

Current trends and future directions in internalized weight stigma research: a scoping review and synthesis of the literature.

BACKGROUND: Since the first papers focused on internalized weight stigma were published in the mid 2000's, the literature has grown into a robust field that complements existing knowledge on weight stigma. Recently, researchers have documented the need for increased conceptual and measurement clarity, to distinguish internalized weight stigma from body dissatisfaction. Although several systematic reviews have been conducted on portions of the internalized weight stigma literature, no review to date has been conducted examining the entirety of the literature. OBJECTIVE: The aim of this research was to conduct a systematic scoping review and synthesis of research on internalized weight stigma. Specifically, we sought to examine the broad scope of the literature, terms used to refer to internalized weight stigma, how internalized weight stigma is defined, sample characteristics, and weight-based framings of internalized weight stigma research. METHODS: We conducted a single-concept search across six databases (EMBASE, Medline, PsychINFO, PubMed, SCOPUS, and Web of Science) of peer-reviewed papers published in English on internalized weight stigma. Data were extracted for article authors, year published, journal name and type, general article topic(s), study design, study location, sample characteristics, variables measured, paper framing, term used to describe internalized weight stigma, and definition of internalized weight stigma. RESULTS: Of the 931 unique records screened, 376 were identified for inclusion in the scoping review. The majority of internalized weight stigma research is characterized by cross-sectional methods, has been conducted in the US, and has utilized samples of higher weight white women. Further, 40 unique terms were used across the literature to refer to internalized weight stigma, and 19 different components of definitions of internalized weight stigma were identified. The literature is also characterized by a focus on understanding the association between internalized weight stigma and health outcomes with an emphasis on obesity. CONCLUSIONS: This scoping review confirms a lack of concept clarity of internalized weight stigma, in part influenced by an inconsistency in definitions of internalized weight stigma across the literature. Considerations are provided for steps to enhance conceptual and measurement clarity. Given the obesity focused framing of much of the research on internalized weight stigma, considerations are also provided for reducing weight-centric approaches to research. In the early 2000's, researchers began to pay more attention to the potential health impacts of believing societal stereotypes, negative attitudes, and beliefs about higher weight people. When these stereotypes, negative attitudes, and beliefs are directed towards the self, it can have significant consequences for an individual's perceptions of self. This research collected and summarized all existing research published in English on internalized weight stigma. Our results highlighted that researchers do not use consistent terminology to refer to internalized weight stigma and that they do not have a consistent definition of internalized weight stigma. Further, a large proportion of the research is focused on obesity or weight loss, which may unintentionally perpetuate weight stigma in scientific research. We provide several recommendations for researchers to address these challenges in future research on internalized weight stigma as well as recommendations to address other identified gaps in the existing literature.

Relationship among group learning, individuals' and groups' internalization of evidence-based practice, and nurses' sustainment of the practice: a cross-sectional study.

PURPOSE: This study, by applying the feedback process of the organizational learning model, examined the relationships among group learning, individuals' and groups' internalization of institutionalized evidence-based practice (I-EBP), and nurses' sustainment of I-EBP. DESIGN/METHODOLOGY/APPROACH: Twelve hospitals were included in this cross-sectional study, with 1,741 nurses from 59 wards. Anonymous questionnaires were administered from October to December 2021. Participants self-reported their wards' group learning, internalization of I-EBP, sustainment of I-EBP, EBP beliefs, intra-hospital transfers, and nursing research experiences. The number of nurses and I-EBP introduction length and type of I-EBP were assessed. Internalization of I-EBP of nurses and groups was considered the mediating variable, while group learning and nurses' sustainment of I-EBP were the independent and dependent variables, respectively. Significant variables in bivariate analyses were used as control variables. Multi-level Mediation Analysis and a significance test of indirect effect using the bootstrap method were conducted. FINDINGS: Responses from 360 nurses in 48 wards from 12 hospitals were analyzed. Groups' internalization of I-EBP significantly mediated the relationship between group learning and nurses' sustainment of I-EBP. In contrast, no significant mediating effect of nurses' internalization of I-EBP was observed. ORIGINALITY/VALUE: In the feedback process of organizational learning, group learning and its subsequent effects on individuals and groups have not been previously examined. Regardless of the nurses' degree of internalization of I-EBP, those who belong to the ward with a high degree of internalization of I-EBP are more likely to sustain it. Conducting group learning may prevent superficial practice, resulting in its sustainability.

Modification of the Antibiotic, Colistin, with Dextrin Causes Enhanced Cytotoxicity and Triggers Apoptosis in Myeloid Leukemia.

Introduction: Acute myeloid leukemia (AML) remains difficult to treat due to its heterogeneity in molecular landscape, epigenetics and cell signaling alterations. Precision medicine is a major goal in AML therapy towards developing agents that can be used to treat patients with different 'subtypes' in combination with current chemotherapies. We have previously developed dextrin-colistin conjugates to combat the rise in multi-drug resistant bacterial infections and overcome dose-limiting nephrotoxicity. Recent evidence of colistin's anticancer activity, mediated through inhibition of intracellular lysine-specific histone demethylase 1 (LSD1/KDM1A), suggests that dextrin-colistin conjugates could be used to treat cancer cells, including AML. This study aimed to evaluate whether dextrin conjugation (which reduces in vivo toxicity and prolongs plasma half-life) could enhance colistin's cytotoxic effects in myeloid leukemia cell lines and compare the intracellular uptake and localization of the free and conjugated antibiotic. Results: Our results identified a conjugate (containing 8000 g/mol dextrin with 1 mol% succinoylation) that caused significantly increased toxicity in myeloid leukemia cells, compared to free colistin. Dextrin conjugation altered the mechanism of cell death by colistin, from necrosis to caspase 3/7-dependent apoptosis. In contrast, conjugation via a reversible ester linker, instead of an amide, had no effect on the mechanism of the colistin-induced cell death. Live cell confocal microscopy of fluorescently labelled compounds showed both free and dextrin-conjugated colistins were endocytosed and co-localized in lysosomes, and increasing the degree of modification by succinoylation of dextrin significantly reduced colistin internalization. Discussion: Whilst clinical translation of dextrin-colistin conjugates for the treatment of AML is unlikely due to the potential to promote antimicrobial resistance (AMR) and the relatively high colistin concentrations required for anticancer activity, the ability to potentiate the effectiveness of an anticancer drug by polymer conjugation, while reducing side effects and improving biodistribution of the drug, is very attractive, and this approach warrants further investigation.

Plasmonics nanorod biosensor for in situ intracellular detection of gene expression biomarkers in intact plant systems.

The intracellular developmental processes in plants, particularly concerning lignin polymer formation and biomass production are regulated by microRNAs (miRNAs). MiRNAs including miR397b are important for developing efficient and cost-effective biofuels. However, traditional methods of monitoring miRNA expression, like PCR, are time-consuming, require sample extraction, and lack spatial and temporal resolution, especially in real-world conditions. We present a novel approach using plasmonics nanosensing to monitor miRNA activity within living plant cells without sample extraction. Plasmonic biosensors using surface-enhanced Raman scattering (SERS) detection offer high sensitivity and precise molecular information. We used the Inverse Molecular Sentinel (iMS) biosensor on unique silver-coated gold nanorods (AuNR@Ag) with a high-aspect ratio to penetrate plant cell walls for detecting miR397b within intact living plant cells. MiR397b overexpression has shown promise in reducing lignin content. Thus, monitoring miR397b is essential for cost-effective biofuel generation. This study demonstrates the infiltration of nanorod iMS biosensors and detection of non-native miRNA 397b within plant cells for the first time. The investigation successfully demonstrates the localization of nanorod iMS biosensors through TEM and XRF-based elemental mapping for miRNA detection within plant cells of Nicotiana benthamiana. The study integrates shifted-excitation Raman difference spectroscopy (SERDS) to decrease background interference and enhance target signal extraction. In vivo SERDS testing confirms the dynamic detection of miR397b in Arabidopsis thaliana leaves after infiltration with iMS nanorods and miR397b target. This proof-of-concept study is an important stepping stone towards spatially resolved, intracellular miRNA mapping to monitor biomarkers and biological pathways for developing efficient renewable biofuel sources.

The laminin receptor is a receptor for Micropterus salmoides rhabdovirus.

Micropterus salmoides rhabdovirus (MSRV) is an important pathogen of largemouth bass. Despite extensive research, the functional receptors of MSRV remained unknown. This study identified the host protein, laminin receptor (LamR), as a cellular receptor facilitating MSRV entry into host cells. Our results demonstrated that LamR directly interacts with MSRV G protein, playing a pivotal role in the attachment and internalization processes of MSRV. Knockdown of LamR with siRNA, blocking cells with LamR antibody, or incubating MSRV virions with soluble LamR protein significantly reduced MSRV entry. Notably, we found that LamR mediated MSRV entry via clathrin-mediated endocytosis. Additionally, our findings revealed that MSRV G and LamR were internalized into cells and co-localized in the early and late endosomes. These findings highlight the significance of LamR as a cellular receptor facilitating MSRV binding and entry into target cells through interaction with the MSRV G protein. IMPORTANCE: Despite the serious epidemic caused by Micropterus salmoides rhabdovirus (MSRV) in largemouth bass, the precise mechanism by which it invades host cells remains unclear. Here, we determined that laminin receptor (LamR) is a novel target of MSRV, that interacts with its G protein and is involved in viral attachment and internalization, transporting with MSRV together in early and late endosomes. This is the first report demonstrating that LamR is a cellular receptor in the MSRV life cycle, thus contributing new insights into host-pathogen interactions.

Unveiling Nanoparticles: Recent Approaches in Studying the Internalization Pattern of Iron Oxide Nanoparticles in Mono- and Multicellular Biological Structures.

Nanoparticle (NP)-based solutions for oncotherapy promise an improved efficiency of the anticancer response, as well as higher comfort for the patient. The current advancements in cancer treatment based on nanotechnology exploit the ability of these systems to pass biological barriers to target the tumor cell, as well as tumor cell organelles. In particular, iron oxide NPs are being clinically employed in oncological management due to this ability. When designing an efficient anti-cancer therapy based on NPs, it is important to know and to modulate the phenomena which take place during the interaction of the NPs with the tumor cells, as well as the normal tissues. In this regard, our review is focused on highlighting different approaches to studying the internalization patterns of iron oxide NPs in simple and complex 2D and 3D in vitro cell models, as well as in living tissues, in order to investigate the functionality of an NP-based treatment.

Biogenic metallic nanoparticles: from green synthesis to clinical translation.

Biogenic metallic nanoparticles (NPs) have garnered significant attention in recent years due to their unique properties and various applications in different fields. NPs, including gold, silver, zinc oxide, copper, titanium, and magnesium oxide NPs, have attracted considerable interest. Green synthesis approaches, utilizing natural products, offer advantages such as sustainability and environmental friendliness. The theranostics applications of these NPs hold immense significance in the fields of medicine and diagnostics. The review explores intricate cellular uptake pathways, internalization dynamics, reactive oxygen species generation, and ensuing inflammatory responses, shedding light on the intricate mechanisms governing their behaviour at a molecular level. Intriguingly, biogenic metallic NPs exhibit a wide array of applications in medicine, including but not limited to anti-inflammatory, anticancer, anti-diabetic, anti-plasmodial, antiviral properties and radical scavenging efficacy. Their potential in personalized medicine stands out, with a focus on tailoring treatments to individual patients based on these NPs' unique attributes and targeted delivery capabilities. The article culminates in emphasizing the role of biogenic metallic NPs in shaping the landscape of personalized medicine. Harnessing their unique properties for tailored therapeutics, diagnostics and targeted interventions, these NPs pave the way for a paradigm shift in healthcare, promising enhanced efficacy and reduced adverse effects.

Chiral silver nanoparticles with surface-anchored L(D)-Cys exhibit dissimilar biological characteristics in vitro but not in vivo.

This work investigated the influence of surface chirality on cellular internalization, cytotoxicity, and tissue distribution of silver nanoparticles (AgNPs). D-cysteine and L-cysteine are chiral forms of the amino acid cysteine. These enantiomers exhibit distinct spatial arrangements, with D-cysteine having a different configuration from L-cysteine. This structural dissimilarity can lead to variations in how these forms interact with biological systems, potentially impacting their cytotoxic responses. Four distinct types of AgNPs were synthesized, each possessing a unique surface coating: pristine AgNPs (pAgNPs), L-cysteine coated AgNPs (AgNPs@L-Cys), D-cysteine coated AgNPs (AgNPs@D-Cys), and racemic AgNPs coated with both L-Cys and D-Cys (AgNPs@L/D-Cys). We found chiral-dependent cytotoxicity of AgNPs on J774A.1 cells. Specifically, AgNPs@L-Cys exhibited the highest toxicity, and AgNPs@D-Cys exhibited the lowest toxicity. Meanwhile, the cellular uptake of the AgNPs correlated nicely with their cytotoxicity, with AgNPs@L-Cys being internalized to the greatest extent while AgNPs@D-Cys displays the least internalization. Scavenger receptors and clathrin predominantly mediate the cellular internalization of these AgNPs. Strikingly, the dissimilar cellular internalization and cytotoxicity of AgNPs with different chirality were eliminated upon protein corona coverage. Notably, following intravenous injection in mice, these four types of AgNPs showed similar patterns among various organs due to the inevitable protein adsorption in the bloodstream. These findings underscored the pivotal role of surface chirality in governing the biological interactions and toxicity of AgNPs.

Surface-Available HER2 Levels Alone Are Not Indicative of Cell Response to HER2-Targeted Antibody-Drug Conjugate Therapies.

HER2-targeting therapies have advanced breast cancer treatment over the past decade. Clinically, eligibility for HER2 therapies is determined by assessing HER2 levels on tumor cell surfaces through immunohistochemistry or by gene regulation through fluorescence in situ hybridization. HER2 therapies are not always effective in patients with elevated levels of HER2, questioning whether the amount of HER2 is sufficiently predictive of patient outcomes. Additionally, the HER2-targeting antibody-drug conjugate (ADC) Enhertu® was recently approved for metastasized HER2-low cancers, confirming the benefits of HER2 treatment for patients with low HER2 levels. To evaluate the correlation between HER2 levels and treatment efficacy, we quantified HER2 on eight cell lines using flow cytometry while simultaneously determining the toxicity of two HER2-targeting ADCs. Both HER2-high cell lines and HER2-low cell lines had significant toxicity responses to ADCs. We quantified HER2 internalization and found no correlation between HER2 levels and the percentage of internalization. We found a useful metric suggesting that a minimum number of HER2 receptors trafficked to lysosomes is sufficient to provide effective treatment. Our results indicate that the current standards of determining eligibility for HER2 therapy could limit patients' access to effective treatment. In conclusion, HER2 levels are not wholly adequate to determine the response to ADC treatment.

Synergistic effects of quercetin-loaded CoFe2O4@Liposomes regulate DNA damage and apoptosis in MCF-7 cancer cells: based on biophysical magnetic hyperthermia.

INTRODUCTION: Breast cancer (BC) is the most common malignancy in women globally. Significant progress has been made in developing structural nanoparticles (NPs) and formulations for targeted smart drug delivery (SDD) of pharmaceuticals, improving the precision of tumor cell targeting in therapy. SIGNIFICANCE: Magnetic hyperthermia (MHT) treatment using magneto-liposomes (MLs) has emerged as a promising adjuvant cancer therapy. METHODS: CoFe2O4 magnetic NPs (MNPs) were conjugated with nanoliposomes to form MLs, and the anticancer drug quercetin (Que) was loaded into MLs, forming Que-MLs composites for antitumor approach. The aim was to prepare Que-MLs for DD systems (DDS) under an alternating magnetic field (AMF), termed chemotherapy/hyperthermia (chemo-HT) techniques. The encapsulation efficiency (EE), drug loading capacity (DL), and drug release (DR) of Que and Que-MLs were evaluated. RESULTS: The results confirmed successful Que-loading on the surface of MLs, with an average diameter of 38 nm and efficient encapsulation into MLs (69%). In vitro, experimental results on MCF-7 breast cells using MHT showed high cytotoxic effects of novel Que-MLs on MCF-7 cells. Various analyses, including cytotoxicity, apoptosis, cell migration, western blotting, fluorescence imaging, and cell membrane internalization, were conducted. The Acridine Orange-ethidium bromide double fluorescence test identified 35% early and 55% late apoptosis resulting from Que-MLs under the chemo-HT group. TEM results indicated MCF-7 cell membrane internalization and digestion of Que-MLs, suggesting the presence of early endosome-like vesicles on the cytoplasmic periphery. CONCLUSIONS: Que-MLs exhibited multi-modal chemo-HT effects, displaying high toxicity against MCF-7 BC cells and showing promise as a potent cytotoxic agent for BC chemotherapy.