Optical lateral flow assays in early diagnosis of SARS-CoV-2 infection.

So far, the 2019 novel coronavirus (COVID-19) is spreading widely worldwide. The early diagnosis of infection by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is essential to provide timely treatment and prevent its further spread. Lateral flow assays (LFAs) have the advantages of rapid detection, simple operation, low cost, ease of mass production, and no need for special devices and professional operators, which make them suitable for self-testing at home. This review focuses on the early diagnosis of SARS-CoV-2 infection based on optical LFAs including colorimetric, fluorescent (FL), chemiluminescent (CL), and surface-enhanced Raman scattering (SERS) LFAs for the detection of SARS-CoV-2 antigens and nucleic acids. The types of recognition components, detection modes used for antigen detection, labels employed in different optical LFAs, and strategies to improve the detection sensitivity of LFAs were reviewed. Meanwhile, LFAs coupled with different nucleic acid amplification techniques and CRISPR-Cas systems for the detection of SARS-CoV-2 nucleic acids were summarized. We hope this review provides research mentalities for developing highly sensitive LFAs that can be used in home self-testing for the early diagnosis of SARS-CoV-2 infection.

A simple chemiluminescent method for the quantification of exosomes based on horseradish peroxidase adsorbed on two-dimensional nanomaterials.

The development of simple methods for the isolation and quantification of exosomes in biological samples is important. By using the typical two-dimensional (2D) nanomaterials, graphene oxide (GO), the present work first studied the interaction of liposomes with the nanocomposites formed by adsorbing HRP on the GO surface and found the presence of liposomes led to the release of HRP from the GO surface to the solution phase triggering the luminol-H2O2 chemiluminescence (CL) reaction to emit light. Benefiting from the similarity of exosomes to liposomes in both composition and morphology aspects, the GO-HRP nanocomposites with a mass ratio of 120:1 and 160:1 were employed for the quantitative detection of exosomes in 100-fold diluted serum samples. The whole detection process took about 15 min and as low as 3.2 × 102 particles µL-1 of exosomes could be sensitively detected. In addition to GO-HRP nanocomposites, the CL responses of other nanocomposites obtained from adsorbing HRP on other 2D nanomaterials such as layered MoS2 for exosomes were also tested. MoS2-HRP exhibited similar behavior and the LODs for the detection of exosomes were 5.8 × 102 particles µL-1. The proposed assays were a biomarker-independent quantitative method that achieved the quantification of exosomes in serum samples directly without an isolation process.

Nisin and ε-polylysine combined treatment enhances quality of fresh-cut jackfruit at refrigerated storage.

This study investigated the effects of nisin combined with ε-polylysine on microorganisms and the refrigerated quality of fresh-cut jackfruit. After being treated with distilled water (control), nisin (0.5 g/L), ε-polylysine (0.5 g/L), and the combination of nisin (0.1 g/L) and ε-polylysine (0.4 g/L), microporous modified atmosphere packaging (MMAP) was carried out and stored at 10 ± 1°C for 8 days. The microorganisms and physicochemical indexes were measured every 2 days during storage. The results indicated that combined treatment (0.1 g/L nisin, 0.4 g/L ε-polylysine) had the best preservation on fresh-cut jackfruit. Compared with the control, combined treatment inhibited microbial growth (total bacterial count, mold and yeast), reduced the weight loss rate, respiratory intensity, polyphenol oxidase and peroxidase activities, and maintained higher sugar acid content, firmness, and color. Furthermore, it preserved higher levels of antioxidant compounds, reduced the accumulation of malondialdehyde and hydrogen peroxide, thereby reducing oxidative damage and maintaining high nutritional and sensory qualities. As a safe application of natural preservatives, nisin combined with ε-polylysine treatment has great application potential in the fresh-cut jackfruit industry.

Melatonin treatment maintains the quality and delays senescence of postharvest cattails (Typha latifolia L.) during cold storage.

Melatonin treatment was investigated for the sensory quality and senescence in postharvest cattails (Typha latifolia L.) during cold storage. The 0.75 mM melatonin treatment reduced surface browning and delaying lignification of Cattails stored at 4 °C. The results showed that melatonin treatment slowed weight loss and firmness, maintained sensory quality and reducing sugar content. Melatonin treatment reduced browning by inhibiting the increase of MDA and H2O2 contents and POD activity. Melatonin treatment maintained high non-enzymatic antioxidant components (Vitamin C and total phenolic content) and antioxidant enzyme activities (SOD, CAT, and APX), thereby alleviating the browning and senescence of postharvest cattails. These findings indicate that melatonin treatment can maintain postharvest cattails quality.

Two-dimensional metal-organic framework catalyzed chemiluminescent reaction for alpha-glucosidase inhibitor screening.

α-Glucosidase inhibitors (AGIs) are oral antidiabetic drugs used in the treatment of type Ⅱ diabetes. It is integral to establish methods for AGIs screening. For the detection of α-glucosidase (α-Glu) activity and screening of AGIs, a chemiluminescence (CL) platform was established based on cascade enzymatic reactions. Firstly, the catalytic activity of a two-dimensional (2D) metal-organic framework (MOF) with iron as central metal atoms and 1,3,5-benzene tricarboxylic acid as a ligand (denoted as 2D Fe-BTC) in the luminol-hydrogen peroxide (H2O2) CL reaction were studied. Mechanism studies showed that the Fe-BTC may react with H2O2 to produce ·OH and act as catalase to facilitate the decomposition of H2O2 to produce O2, thus showing good catalytic activity in the luminol-H2O2 CL reaction. The proposed luminol-H2O2-Fe-BTC CL system exhibited an outstanding response to glucose with the aid of glucose oxidase (GOx). The luminol-GOx-Fe-BTC system showed a detection linear range from 50 nM to 10 µM with a detection limit (LOD) of 3.62 nM for glucose detection. Then, the luminol-H2O2-Fe-BTC CL system was applied to the detection of α-glucosidase (α-Glu) activity and screening of AGIs based on cascade enzymatic reactions using acarbose and voglibose as model drugs. The IC50 of acarbose and voglibose was 7.39 µM and 1.89 mM, respectively.

A turn-on chemiluminescent assay for alkaline phosphatase using two-dimensional Fe-centered metal-organic frameworks as the signaling probe.

Alkaline phosphatase (ALP) is an essential enzyme involved in cell phosphorus metabolism. Developing sensitive and accurate ALP quantitative assays is significant. In this study, a turn-on chemiluminescence (CL) analysis platform for the detection of ALP activity in human serum was established based on two-dimensional (2D) Fe-centered metal-organic frameworks with 1,3,5-benzene tricarboxylic acid as ligands (denoted as 2D Fe-BTC). The 2D Fe-BTC as the signaling probe reacts with ascorbic acid forming reduced Fe-BTC which catalyzes the luminol CL reaction producing a strong CL signal. The 2D Fe-BTC-based luminol CL reaction exhibited good CL responses when the concentration of ascorbic acid was in the range of 5-500 nM. By employing magnesium ascorbyl phosphate (MAP) as the substrate which can be hydrolyzed by ALP to generate ascorbic acid, a turn-on CL assay for the detection of ALP was established. Under optimal conditions, as low as 0.00046 U L-1 of ALP could be sensitively detected with a linear range of 0.001-0.1 U L-1. ALP in human serum can be detected after a simple dilution process without any other pretreatment.

Ultrasensitive colorimetric detection of tetracyclines based on in-situ growth of gold nanoflowers.

A colorimetric method based on in-situ generation of gold nanoflowers for the detection of tetracyclines (TCs) was proposed. We found that gold nanoflowers could be formed in the HAuCl4-NH2OH redox reaction directly without the addition of small-sized gold nanoparticles (Au NPs) as seeds when an alkaline borax buffer solution was employed as the reaction medium. Interestingly, the shape and size of the generated gold nanoflowers were regulated with TC. Briefly, large flower-like gold nanoparticles were formed with a low concentration of TC while small spherical gold nanoparticles were generated with a high concentration of TC. The generated gold nanoflowers exhibited different surface plasmon absorption (SPR) properties. Thus, a simple and rapid colorimetric method was established for the detection of TC antibiotics. This method exhibited high sensitivity for the detection of TC, oxytetracycline (OTC), and doxycycline (DC) with detection limits of 2.23 nM, 1.19 nM, and 5.81 nM, respectively. The proposed colorimetric method was applied to the determination of TC in both milk samples and water samples.

Analysis of the related factors of atypical squamous cells of undetermined significance (ASC-US) in cervical cytology of post-menopausal women.

Introduction: Atrophy of the reproductive tract mucosa caused by the decrease of estrogen may increase the detection rate of ASC-US in cervical cytology of post-menopausal women. In addition, other pathogenic infections and inflammation can change the cellular morphology and increase the detection rate of ASC-US. However, further studies are needed to elucidate whether the high detection rate of ASC-US in post-menopausal women leads to the high referral rate of colposcopy. Methods: This retrospective study was conducted to document ASC-US in cervical cytology reports at the Department of Cytology at Gynecology and Obstetrics, Tianjin Medical University General Hospital between January 2006 and February 2021. We then analyzed 2,462 reports of women with ASC-US at the Cervical Lesions Department. A total of 499 patients with ASC-US and 151 cytology with NILM participants underwent vaginal microecology tests. Results: The average reporting rate of ASC-US in cytology was 5.7%. The detection rate of ASC-US in women aged > 50 years (7.0%) was significantly higher than that in women aged ≤50 years (5.0%) (P<0.05). The CIN2+ detection rate was significantly lower in the post- (12.6%) than in pre-menopausal (20.5%) patients with ASC-US (P <0.05). The prevalence of abnormal reporting rate of vaginal microecology was significantly lower in the pre-menopausal group (56.2%) than that in the post-menopausal group (82.9%) (P<0.05). The prevalence of bacterial vaginosis (BV) (19.60%) was relatively high in the pre-menopausal group, but the abundance of bacteria-inhibiting flora (40.79%) was mainly an abnormality in the post-menopausal group. The vaginal microecological abnormality rate of the women with HR-HPV (-) of ASC-US was 66.22%, which was significantly higher than that of the HR-HPV (-) and the NILM group (52.32%; P<0.05). Discussion: The detection rate of ASC-US in women aged > 50 years was higher than that ≤50 years, but the detection rate of CIN2+ was lower in the post-menopausal women with ASC-US. However, vaginal microecological abnormalities may increase the false-positive diagnosis rate of ASC-US. The vaginal microecological abnormalities of the menopausal women with ASC-US are mainly attributed to infectious diseases such as BV, and it mainly occurs in the post-menopausal women was bacteria-inhibiting flora. Therefore, to avoid the high referral rate for colposcopy, more attention should be paid to the detection of vaginal microecology.

Interactions between microbiota and cervical epithelial, immune, and mucus barrier.

The female reproductive tract harbours hundreds of bacterial species and produces numerous metabolites. The uterine cervix is located between the upper and lower parts of the female genital tract. It allows sperm and birth passage and hinders the upward movement of microorganisms into a relatively sterile uterus. It is also the predicted site for sexually transmitted infection (STI), such as Chlamydia, human papilloma virus (HPV), and human immunodeficiency virus (HIV). The healthy cervicovaginal microbiota maintains cervical epithelial barrier integrity and modulates the mucosal immune system. Perturbations of the microbiota composition accompany changes in microbial metabolites that induce local inflammation, damage the cervical epithelial and immune barrier, and increase susceptibility to STI infection and relative disease progression. This review examined the intimate interactions between the cervicovaginal microbiota, relative metabolites, and the cervical epithelial-, immune-, and mucus barrier, and the potent effect of the host-microbiota interaction on specific STI infection. An improved understanding of cervicovaginal microbiota regulation on cervical microenvironment homeostasis might promote advances in diagnostic and therapeutic approaches for various STI diseases.

Leveraging 16S rRNA data to uncover vaginal microbial signatures in women with cervical cancer.

Microbiota-relevant signatures have been investigated for human papillomavirus-related cervical cancer (CC), but lack consistency because of study- and methodology-derived heterogeneities. Here, four publicly available 16S rRNA datasets including 171 vaginal samples (51 CC versus 120 healthy controls) were analyzed to characterize reproducible CC-associated microbial signatures. We employed a recently published clustering approach called VAginaL community state typE Nearest CentroId clAssifier to assign the metadata to 13 community state types (CSTs) in our study. Nine subCSTs were identified. A random forest model (RFM) classifier was constructed to identify 33 optimal genus-based and 94 species-based signatures. Confounder analysis revealed confounding effects on both study- and hypervariable region-associated aspects. After adjusting for confounders, multivariate analysis identified 14 significantly changed taxa in CC versus the controls (P < 0.05). Furthermore, predicted functional analysis revealed significantly upregulated pathways relevant to the altered vaginal microbiota in CC. Cofactor, carrier, and vitamin biosynthesis were significantly enriched in CC, followed by fatty acid and lipid biosynthesis, and fermentation of short-chain fatty acids. Genus-based contributors to the differential functional abundances were also displayed. Overall, this integrative study identified reproducible and generalizable signatures in CC, suggesting the causal role of specific taxa in CC pathogenesis.

Which is the best management for women with normal cervical cytologic findings despite positivity for non-16/18 high risk human papillomaviruses?

Women who test positive for the human papillomavirus (HPV) but have normal cytology constitute the predominant subgroup of patients in the screening population in the post-vaccination era. The distribution of HPV genotypes changed dramatically, which was attributable to an increase in HPV vaccination coverage. These changes have created uncertainty about how to properly manage women with normal cytology, non-HPV16/18 infections, or persistent infections. Current recommendations include retesting and continued surveillance in the absence of HPV16/18 infection. However, these are not always applicable. The ability to implement genotyping or incorporate HPV16/18 with some additional high-risk HPV (HR-HPV) types for triage and management with the aim of identifying type-specific risks in this population could be acceptable. When the next set of guidelines is updated, generating potential triage strategies for detecting high-grade cervical lesions, such as the p16/Ki67 cytology assay and other alternatives that incorporate genotyping with newer tests, should be considered. Current clinical management is shifting to risk-based strategies; however, no specific risk threshold has been established in this population. Importantly, innovative triage testing should be evaluated in combination with primary screening and management. Furthermore, there is an untapped opportunity to coordinate HPV genotyping in combination with colposcopic characteristics to modify risk in this group. Hence, providing a more personalized schedule through the efficient application of risk stratification and improving the detection of pre-cancer and cancer is an option worth exploring.

Disturbances of Vaginal Microbiome Composition in Human Papillomavirus Infection and Cervical Carcinogenesis: A Qualitative Systematic Review.

Background: Emerging evidence has demonstrated a close association between perturbations in vaginal microbiota composition in women and human papillomavirus (HPV) infection, cervical lesions, and cervical cancer (Ca); however, these findings are highly heterogeneous and inconclusive. Aim: To perform a comprehensive systematic review of the global disturbance in the vaginal microbiota, specifically in women with HPV-associated cervical diseases, and to further conduct within- and across-disease comparisons. Method: Twenty-two records were identified in a systematic literature search of PubMed, Web of Science, and Embase up to February 28, 2022. We extracted microbial changes at the community (alpha and beta diversity) and taxonomic (relative abundance) levels. Within- and across-disease findings on the relative abundance of taxonomic assignments were qualitatively synthesized. Results: Generally, significantly higher alpha diversity was observed for HPV infection, cervical lesions, and/or cancer patients than in controls, and significant differences within beta diversity were observed for the overall microbial composition across samples. In within-disease comparisons, the genera Gardnerella, Megasphaera, Prevotella, Peptostreptococcus, and Streptococcus showed the greatest abundances with HPV infection; Sneathia and Atopobium showed inconsistent abundance with HPV infection, and Staphylococcus was observed in Ca. Across diseases, we find increased levels of Streptococcus and varying levels of Gardnerella were shared across HPV infections, high-grade squamous intraepithelial lesions, and Ca, whereas Lactobacillus iners varied depending on the HPV-related disease subtype. Conclusions: This systematic review reports that vaginal microbiome disturbances are correlated to the depletion of Lactobacillus, enrichment of anaerobes, and increased abundance of aerobic bacteria in HPV infection and related cervical diseases. Moreover, L. iners may exert either protective or pathogenic effects on different HPV-related diseases.

Enhancement effect of 2, 3-dimethyl maleic acid on luminol chemiluminescence reactions and its application in detection of sequence-specific DNA related to hepatitis B virus.

2, 3-dimethyl maleic acid (DMMA) was found to enhance luminol-H2O2 chemiluminescent (CL) reactions, among which the strongest enhancement effect was observed by using polyethyleneimine-templated gold nanoclusters (PEI-Au NCs) as the catalyst. With the addition of DMMA, the CL signal of the PEI-Au NCs-catalyzed luminol-H2O2 reaction enhanced about 630-fold, and a flash-type CL profile was obtained. Mechanism studies showed that the luminophore was still 3-aminophthalate anions in the excited state (3-APA*), and superoxide radical (O2·-) played an important role during the CL process. Under the optimized experimental conditions, the lowest concentration of PEI-Au NCs can be detected was 0.168 nM which was 82-fold lower than that without an enhancer. Furthermore, the catalytic activity of biotinylated PEI-Au NCs in the DMMA-enhanced luminol system was similar to PEI-Au NCs, providing a good opportunity for the development of CL bioanalysis platforms using PEI-Au NCs as the label. Thus, the DMMA-enhanced luminol-H2O2 system was applied to the CL detection of sequence-specific DNA related to the hepatitis B virus (HBV) using PEI-Au NCs as the label. The CL platform exhibited linearly enhanced CL response with the increasing amount of target DNA ranging from 0.0025 to 0.5 pmol. As low as 0.002 pmol of HBV DNA could be sensitively detected, which was superior to the previously reported methods.

Colorimetric assay for tetracyclines based on europium ion-induced aggregation of gold nanoparticles.

Oxytetracycline-capped gold nanoparticles (OTC-Au NPs) were prepared using sodium borohydride as the reductant and OTC as the capping agent, respectively. The prepared OTC-Au NPs with a size of 6 nm have a maximum surface plasma resonance (SPR) absorption located at 514 nm. The OTC on the surface of Au NPs still can coordinate with Eu3+ ions. Due to the property that OTC has multivalent binding sites with Eu3+ ions, Eu3+ ions can induce the aggregation of OTC-Au NPs. Based on the Eu3+ ions-aggregated OTC-Au NPs, a simple aptamer-free colorimetric sensing method for TCs was developed. Briefly, free TCs compete with OTC on the surface of Au NPs resulting in the change of OTC-Au NPs from an aggregation state to a dispersed state. The whole process takes only 5 min, and as low as 20 nM OTC, 14 nM tetracycline (TC), and 20 nM doxycycline (DC) could be sensitively detected, respectively. The proposed method was also featured as good repeatability and specificity, and was applied to the detection of OTC in lake water with satisfactory recovery.

Aerobic Vaginitis Diagnosis Criteria Combining Gram Stain with Clinical Features: An Establishment and Prospective Validation Study.

Wet-mount microscopy aerobic vaginitis (AV) diagnostic criteria need phase-contrast microscopy and keen microscopists, and the preservation of saline smears is less common in clinical practice. This research work developed new AV diagnostic criteria that combine Gram stain with clinical features. We enrolled 325 AV patients and 325 controls as a study population to develop new AV diagnostic criteria. Then, an independent group, which included 500 women, was used as a validation population. AV-related microscopic findings on Gram-stained and wet-mount smears from the same participants were compared. The accuracy of bacterial indicators from the two methods was verified by bacterial 16S rRNA V4 sequencing (n = 240). Logistic regression was used to analyse AV-related clinical features. The screened clinical features were combined with Gram-stain microscopic indicators to establish new AV diagnostic criteria. There were no significant differences in the leukocyte counts or the parabasal epitheliocytes (PBC) proportion between the Gram-stain and wet-mount methods (400×). Gram stain (1000×) satisfied the ability to identify bacteria as verified by 16S rRNA sequencing but failed to identify toxic leukocytes. The new criteria included: Lactobacillary grades (LBG) and background flora (Gram stain, 1000×), leukocytes count and PBC proportion (Gram stain, 400×), and clinical features (vaginal pH > 4.5, vagina hyperemia, and yellow discharge). These criteria satisfied the accuracy and reliability for AV diagnosis (Se = 86.79%, Sp = 95.97%, and Kendall's W value = 0.899) in perspective validation. In summary, we proposed an alternative and valuable AV diagnostic criteria based on the Gram stain, which can make it possible to diagnose common vaginitis like AV, BV, VVC, and mixed infections on the same smear and can be available for artificial intelligence diagnosis in the future.

The pathogenesis of prevalent aerobic bacteria in aerobic vaginitis and adverse pregnancy outcomes: a narrative review.

BACKGROUND: Aerobic vaginitis is a common cause of vaginal discharge in reproductive-age women, increasing the risk of negative pregnancy outcomes such as premature delivery, abortion, premature rupture of membranes and stillbirth. However, the aetiology and pathogenesis of aerobic vaginitis causing negative pregnancy outcomes are still unclear, and there is no unified and standardized treatment method for aerobic vaginitis in the pregnancy period. METHODS: We conducted a literature search of published studies in the English language focusing on aerobic vaginitis and its association with adverse pregnancy outcomes utilizing PubMed and Web of Science from January 1973 through June 2021. The common pathogenic bacteria of aerobic vaginitis during pregnancy, such as group B Streptococcus, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Klebsiella pneumoniae, as well as the related adverse pregnancy outcomes and existing treatments were reviewed. RESULTS: A total of 4534 articles were identified, and 97 studies that had inclusion criteria were subjected to careful review. The pathogenic bacteria of aerobic vaginitis can produce different toxins or affect the local immunity of patients and then lead to the occurrence of infection. Fresh wet mount microscopy is the preferred diagnostic method for aerobic vaginitis. Clindamycin is a common antibiotic used for aerobic vaginitis in pregnant women. The use of products combining probiotics has achieved excellent treatment success. CONCLUSIONS: Future research in this field can provide insights regarding the mechanism of aerobic vaginitis-induced adverse pregnancy outcomes in humans and ways to prevent their occurrence.

Aerobic vaginitis is an infection of the vagina that increases the risk of negative pregnancy outcomes. The aetiology and pathogenesis of aerobic vaginitis causing negative pregnancy outcomes are still unclear. This paper reviews the common pathogenic bacteria of aerobic vaginitis during pregnancy, and the related adverse pregnancy outcomes. We also review the existing treatment. Currently, it is believed that the microflora in aerobic vaginitis is composed of commensal aerobic microorganisms of intestinal origin, and the most frequently encountered bacteria are group B Streptococcus, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis and Klebsiella pneumoniae. The pathogenic bacteria of aerobic vaginitis can produce different toxins or affect the local immunity of patients and then lead to the occurrence of infection. Fresh wet mount microscopy is the preferred diagnostic method for aerobic vaginitis. Clindamycin is a common antibiotic used for aerobic vaginitis in pregnant women. The use of products combining probiotics has achieved excellent treatment success. This study provides a reference for future research and early diagnosis and treatment during pregnancy. Future research in this field can provide insights regarding the mechanisms of aerobic vaginitis-induced adverse pregnancy outcomes in humans and ways to prevent their occurrence.

Recent Advances in Presentation, Diagnosis and Treatment for Mixed Vaginitis.

Mixed vaginitis is the simultaneous presence of at least two types of vaginitis, contributing to an abnormal vaginal milieu and leading to vaginal symptoms and signs. However, associations between symptoms and the type of mixed vaginitis have not been clearly elucidated, and research on mixed vaginitis is still in the preliminary stage. Therefore, the pathogenic mechanism of mixed vaginitis remains understudied. Mixed vaginitis generally involves the formation of mixed biofilms. The study of polymicrobial interactions and mixed biofilms will provide a new idea for the understanding of mixed vaginitis. Moreover, this review summarizes some effective management and laboratory diagnosis of mixed vaginitis to avoid inappropriate therapy, recurrence, and reinfection. It is of high clinical importance to obtain relevant clinical data to improve clinical knowledge about mixed vaginitis.

A Simple Colorimetric Analytical Assay for the Determination of Tetracyclines Based on In-situ Generation of Gold Nanoparticles Coupling with a Gold Staining Technique.

The development of simple and sensitive detection methods for tetracyclines (TCs) is crucial for their routine detection. The present study developed a colorimetric method for the detection of TCs based on the in-situ generation of AuNPs, which were subsequently coupled with a gold staining reaction. Briefly, TCs containing phenolic groups reduce HAuCl4 to form gold nanoparticles (AuNPs) as gold seeds. In the gold staining process, the gold seeds catalyze the reduction of HAuCl4 by NH2OH to form gold atoms that deposit on the surface of AuNPs, resulting in the enlargement of AuNPs. Sensitive detection of TCs was achieved by employing the gold staining technique. As low as 14, 18.9, and 1.98 nM of oxytetracycline (OTC), tetracycline (TC), and doxycycline (DC), respectively, could be sensitively detected. The proposed method also exhibited good repeatability and specificity, and then was applied to the determination of OTC in milk samples.

Electron-Accepting Micelles Deplete Reduced Nicotinamide Adenine Dinucleotide Phosphate and Impair Two Antioxidant Cascades for Ferroptosis-Induced Tumor Eradication.

Ferroptotic antitumor therapy has been compromised by various intracellular antioxidants, particularly glutathione and thioredoxin. Both are cofactors of glutathione peroxide 4 (GPX4) that act against oxidative stress via catalyzing the reduction of lipid peroxides. It was postulated that tailored polymer micelles could enhance ferroptotic antitumor efficacy via diminishing glutathione and thioredoxin under hypoxia. The aim was to engineer hypoxia-responsive micelles for selective enhancement of ferroptotic cell death in solid tumor. The polymer contains hydrophilic poly(ethylene glycol) (PEG) that is linked by azobenzene linker with nitroimidazole-conjugated polypeptide. The tailored polymer could self-assemble into nanoscale micelles to encapsulate RAS-selective lethal small molecule 3, a covalent GPX4 inhibitor. Under hypoxia, the azobenzene moiety enabled PEG shedding and enhanced micelles uptake in 4T1 cells. Likewise, the nitroimidazole moiety was reduced by the overexpressed nitroreductase with reduced nicotinamide adenine dinucleotide phosphate (NADPH) as the cofactor, resulting in transient depletion of NADPH. This impaired both the glutathione and thioredoxin redox cycle, leading to diminished intracellular glutathione and thioredoxin. The selective potency of ferroptotic micelles in depleting NADPH, glutathione and thioredoxin was further verified in vivo in the 4T1 tumor xenograft mice model. This work highlights the role of hypoxia-responsive polymers in enhancing the potency of ferroptotic inducers against solid tumors without additional side effects to healthy organs.

Nanomaterial-enhanced chemiluminescence reactions and their applications.

Chemiluminescence (CL) analysis is a trace analytical method that possesses advantages including high sensitivity, wide linear range, easy operation, and simple instruments. With the development of nanotechnology, many nanomaterial (NM)-enhanced CL systems have been established in recent years and applied for the CL detection of metal ions, anions, small molecules, tumor markers, sequence-specific DNA, and RNA. This review summarizes the research progress of the nanomaterial-enhanced CL systems the past five years. These CL reactions include luminol, peroxyoxalate, lucigenin, ultraweak CL reactions, and so on. The CL mechanisms of the nanomaterial-enhanced CL systems are discussed in the first section. Nanomaterials take part in the CL reactions as the catalyst, CL emitter, energy acceptor, and reductant. Their applications are summarized in the second section. Finally, the challenges and opportunities are discussed.