Fluorescein Research Articles

Covalent Organic Framework Functionalized with Fluorescein Isothiocyanate for the Selective and Sensitive Detection of Toxic Mycotoxin

Covalent Organic Framework Functionalized with Fluorescein Isothiocyanate for the Selective and Sensitive Detection of Toxic Mycotoxin

#794 LncRNA NEAT1 protects uremic toxin-induced intestinal epithelial barrier injury by regulating miR-122-5p/Occludin axis

Abstract Background and Aims Long non-coding RNA(LncRNA) has been reported to be associated with intestinal barrier damage. The aim of this study was to explore the mechanism of lncRNA Nuclear enriched abundant transcript 1 (NEAT1) in uremic toxin-induced intestinal epithelial barrier injury. Method Human colon cancer cells (Caco-2) were used to establish intestinal epithelial injury models with the urea treatment in different conditions. Cell Counting Kit-8 (CCK-8) and Western blot screening the best concentration and time. The expressions of lncRNA NEAT1 and miR-122-5p were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Western blot and immunofluorescence were used to detect the expression of tight junction proteins Occludin, ZO-1 and Claudin-1. Sodium fluorescein was used to detect the paracellular permeability of intestinal epithelial injury models. The binding of miR-122-5p to lncRNA NEAT1 and Occludin was determined by bioinformatics analysis and dual luciferase reporter assay. Results The best condition for the injury model was urea treatment in 144 mg/dl for 48 hours. With the increase of urea intervention time and concentration, the damage degree of intestinal epithelial cells is aggravated. Based on the qRT-PCR results, lncRNA NEAT1 was significantly down-regulated in the model group. Meanwhile, the tight junction proteins Occludin, ZO-1 and Claudin-1 were significantly reduced. The permeability of sodium fluorescein was significantly increased in the model group. Overexpression of lncRNA NEAT1 can alleviate the above performances. As the target gene of lncRNA NEAT1, miR-122-5p is significantly up-regulated in the model group. The dual luciferase reporter assay proved that miR-122-5p was targets to Occludin. The protective effect of overexpression lncRNA NEAT1 on intestinal epithelial barrier function is reversed by miR-122-5p mimics. Conclusion LncRNA NEAT1 protects uremic toxin-induced intestinal epithelial barrier injury by regulating miR-122-5p/Occludin axis.

Retention time of different ophthalmic viscosurgical devices during phacoemulsification in rabbit ocular model: A comparative analysis.

To compare retention times of various ophthalmic viscosurgical devices (OVDs) and soft-shell combinations. Experimental study. Eighteen rabbit eyes were divided into six groups of three eyes, based on OVDs tested. A: Endocoat, B: HealonPro, C: Viscoat, D: Provisc, E: Endocoat and HealonPro and F: Viscoat and Provisc. OVDs were stained with 10% fluorescein dye before being injected into the anterior chamber. Phacoemulsification, using fixed parameters, was performed. If OVD persisted after 60 s, a standardised irrigation and aspiration technique replaced phacoemulsification. The time until central and complete clearance of the OVD were video-recorded and measured. Mean central retention times (CRT) were found to be: HealonPro-5.33 ± 2.56 s, Provisc-3.33 ± 1.11 s, Endocoat-75.0 ± 3.26 s, Viscoat-62.33 ± 5.19 s, combining HealonPro with Endocoat-22.67 ± 4.75 s and Provisc with Viscoat-11.0 ± 0.82 s. The mean total retention time (TRT) was: Endocoat-80.0 ± 8.17 s, Viscoat-81.67 ± 2.09 s, HealonPro with Endocoat-81.33 ± 3.35 s, and Provisc with Viscoat-71.0 ± 2.94 s. For HealonPro and Provisc, CRT and TRT remained identical across all trials. Retention times varied, with cohesive OVDs exhibiting shorter durations than dispersive OVDs. Among dispersive OVDs, TRTs were comparable; however, Endocoat displayed an extended CRT. In soft shell trials, the combination of HealonPro and Endocoat exhibited prolonged CRT and TRT, suggesting enhanced corneal protection.

Nanolignin-containing cellulose nanofibrils (LCNF)-enabled multifunctional ratiometric fluorescent bio-nanocomposite films for food freshness monitoring

Herein, the nanolignin-containing cellulose nanofibrils (LCNF)-enabled ratiometric fluorescent bio-nanocomposite film is developed. Interestingly, the inclusion of LCNF in the cellulose-based film enhances the detecting performance of food freshness, such as high sensitivity to biogenic amines (BAs) (limit of detection (LOD) of up to 1.83 ppm) and ultrahigh discernible fluorescence color difference (ΔE = 113.11). The underlying mechanisms are the fluorescence resonance energy transfer (FRET), π - π interaction, and cation – π interaction between LCNF and fluorescein isothiocyanate (FITC), as well as the increased hydrophobicity due to lignin, which increases the interactions of amines with FITC. Its color stability (up to 28 days) and mechanical property (49.4 Mpa) are simultaneously improved. Furthermore, a smartphone based detecting platform is developed to achieve access to food safety. This work presents a novel technology, which can have a great potential in the field of food packaging and safety.

Fluorescent Composite Nano‐ and Microparticles Based on Xanthene Dyes and Iron Oxides

AbstractModern industry actively uses metal structural materials to create innovative devices and components. In order to prevent emergency situations in production, nondestructive testing methods are used to detect defects on the surface of the material being developed. Among the methods of nondestructive testing, one can highlight the method of luminescent magnetic flaw detection, which allows identifying defects on the surface and under the surface of the test sample with minimal effort. It is known that in some cases, the magnetic‐powder method cannot achieve the required sensitivity due to the shape and location of defects themselves or due to certain conditions of object operation. To increase the visibility and sensitivity, magnetic nano‐ and microparticles can be conjugated with the dyes. Iron oxide nanoparticles (Fe3O4, γ‐Fe2O3) can be used as such magnetic particles, since they are quite easy to obtain and have low toxicity, high chemical stability, and a wide variety of magnetic properties. To ensure fluorescence of magnetic particles, a wide variety of fluorescent dyes associated with such particles are used. The dyes can include morin, naphthalimide derivatives, as well as semiconductor quantum dots. This article provides a review of fluorescent composite micro‐ and nanoparticles based on xanthene dyes (rhodamine, fluorescein, and pyronine). Such dyes are economically available luminophores, have good binding to the surface of magnetic particles, and therefore can be suitable for luminescent magnetic flaw detection. This review article discusses methods for producing composite nano‐ and microparticles of iron oxides with xanthene dyes and provides prospects of using composite nano‐ and microparticles in nondestructive testing methods.

Development of Peptide Displacement Assays to Screen for Antagonists of DDB1 Interactions.

The DNA damage binding protein 1 (DDB1) is an essential component of protein complexes involved in DNA damage repair and the ubiquitin-proteasome system (UPS) for protein degradation. As an adaptor protein specific to Cullin-RING E3 ligases, DDB1 binds different receptors that poise protein substrates for ubiquitination and subsequent degradation by the 26S proteasome. Examples of DDB1-binding protein receptors are Cereblon (CRBN) and the WD-repeat containing DDB1- and CUL4-associated factors (DCAFs). Cognate substrates of CRBN and DCAFs are involved in cancer-related cellular processes or are mimicked by viruses to reprogram E3 ligases for the ubiquitination of antiviral host factors. Thus, disrupting interactions of DDB1 with receptor proteins might be an effective strategy for anticancer and antiviral drug discovery. Here, we developed fluorescence polarization (FP)-based peptide displacement assays that utilize full-length DDB1 and fluorescein isothiocyanate (FITC)-labeled peptide probes derived from the specific binding motifs of DDB1 interactors. A general FP-based assay condition applicable to diverse peptide probes was determined and optimized. Mutagenesis and biophysical analyses were then employed to identify the most suitable peptide probe. The FITC-DCAF15 L49A peptide binds DDB1 with a dissociation constant of 68 nM and can be displaced competitively by unlabeled peptides at sub-μM to low nM concentrations. These peptide displacement assays can be used to screen small molecule libraries to identify novel modulators that could specifically antagonize DDB1 interactions toward development of antiviral and cancer therapeutics.

Modulation of the Blood-Brain Barrier by Sigma-1R Activation.

Sigma non-opioid intracellular receptor 1 (Sigma-1R) is an intracellular chaperone protein residing on the endoplasmic reticulum at the mitochondrial-associated membrane (MAM) region. Sigma-1R is abundant in the brain and is involved in several physiological processes as well as in various disease states. The role of Sigma-1R at the blood-brain barrier (BBB) is incompletely characterized. In this study, the effect of Sigma-1R activation was investigated in vitro on rat brain microvascular endothelial cells (RBMVEC), an important component of the blood-brain barrier (BBB), and in vivo on BBB permeability in rats. The Sigma-1R agonist PRE-084 produced a dose-dependent increase in mitochondrial calcium, and mitochondrial and cytosolic reactive oxygen species (ROS) in RBMVEC. PRE-084 decreased the electrical resistance of the RBMVEC monolayer, measured with the electric cell-substrate impedance sensing (ECIS) method, indicating barrier disruption. These effects were reduced by pretreatment with Sigma-1R antagonists, BD 1047 and NE 100. In vivo assessment of BBB permeability in rats indicates that PRE-084 produced a dose-dependent increase in brain extravasation of Evans Blue and sodium fluorescein brain; the effect was reduced by the Sigma-1R antagonists. Immunocytochemistry studies indicate that PRE-084 produced a disruption of tight and adherens junctions and actin cytoskeleton. The brain microcirculation was directly visualized in vivo in the prefrontal cortex of awake rats with a miniature integrated fluorescence microscope (aka, miniscope; Doric Lenses Inc.). Miniscope studies indicate that PRE-084 increased sodium fluorescein extravasation in vivo. Taken together, these results indicate that Sigma-1R activation promoted oxidative stress and increased BBB permeability.

The effect of different irrigation activation methods on the penetration of the irrigation solution into the dentinal tubules in teeth with calcium hydroxide applied and oval-shaped canals: Confocal laser scanning microscope analysis.

This study aimed to evaluate the efficacy of different irrigation techniques including standard needle irrigation (SNI), passive ultrasonic irrigation (PUI), EndoActivator (EA), XP-Endo Finisher (XPF), photon-induced photoacoustic streaming (PIPS), and shock wave enhanced emission photo-acoustic streaming (SWEEPS) systems on the penetration of irrigation solutions into dentinal tubules in teeth with calcium hydroxide (CH) applied and oval-shaped canals by confocal laser scanning microscope (CLSM). Ninety mandibular incisor teeth with oval-shaped canals were included in this study. After the preparation of teeth using Resiproc Blue R25 (VDW, Munich, Germany) canals were filled with a paste based on CH. The teeth were stored in 100% humidity at a temperature of 37°C for 14 days. According to the irrigation activation systems, the teeth were divided into six groups (n = 15); SNI, PUI, EA, XPF, PIPS, and SWEEPS. In each group, 3 irrigation/activation cycles of 20 s of irrigation and 20 s of activation were performed with the relevant activation method. The root canals were filled with fluorescein sodium (Sigma, Alldrich Co., St. Louis, MO, USA) and activated with the relevant activation method for 30 s. Specimens were sectioned horizontally to 1 ± 0.1 mm at 2, 5, and 8 mm from the apex and then examined under the CLSM. Maximum penetration depth, maximum penetration area and penetration percentage were measured by using Image J software. Data were analyzed using a two-way analysis of variance (ANOVA) and posthoc Tukey tests (p = .05). No difference was found between the activation systems in terms of maximum penetration depth and maximum penetration area of irrigation solutions in the apical section (p > .05). The penetration percentage of irrigation solutions was higher in PUI and PIPS compared with EA and XPF at the apical section (p < .05). No difference was found between SNI, PUI, PIPS, and SWEEPS in terms of the penetration percentage of irrigation solutions in all sections (p > .05). The penetration percentage of irrigation solutions was higher in the middle and coronal sections compared to the apical section in EA and XPF (p < .05). Tubule penetration of irrigation solutions in SNI was similar between sections. The Tubule penetration area of irrigation solutions in PUI, EA, XPF, PIPS, and SWEEPS was lower in the apical section than in the middle and coronal sections. RESEARCH HIGHLIGHTS: The tubule penetration depth and area of the irrigation solutions were similar between the activation systems tested in the apical sections. The Tubule penetration area of irrigation solutions in middle and coronal sections was higher in PUI and PIPS than in SNI.

Hyperglycaemia perturbs blood-brain barrier integrity through its effects on endothelial cell characteristics and function

ABSTRACT Breakdown of blood-brain barrier (BBB) represents a key pathology in hyperglycemia-mediated cerebrovascular damage after an ischemic stroke. As changes in the level and nature of vasoactive agents released by endothelial cells (ECs) may contribute to BBB dysfunction, this study first explored the specific impact of hyperglycemia on EC characteristics and secretome. It then assessed whether secretome obtained from ECs subjected to normoglycaemia or hyperglycemia might regulate pericytic cytokine profile differently. Using a triple cell culture model of human BBB, composed of brain microvascular EC (BMEC), astrocytes and pericytes, this study showed that exposure to hyperglycemia (25 mM D-glucose) for 72 h impaired the BBB integrity and function as evidenced by decreases in transendothelial electrical resistance and increases in paracellular flux of sodium fluorescein. Dissolution of zonula occludens-1, a tight junction protein, and appearance of stress fibers appeared to play a key role in this pathology. Despite elevations in angiogenin, endothelin-1, interleukin-8 and basic fibroblast growth factor levels and a decrease in placental growth factor levels in BMEC subjected to hyperglycemia vs normoglycaemia (5.5 mM D-glucose), tubulogenic capacity of BMECs remained similar in both settings. Similarly, pericytes subjected to secretome obtained from hyperglycemic BMEC released higher quantities of macrophage migration inhibitory factor and serpin and lower quantities of monocyte chemoattractant protein-1, intercellular adhesion molecule, interleukin-6 and interleukin-8. Taken together these findings indicate the complexity of the mechanisms leading to BBB disruption in hyperglycemic settings and emphasize the importance of endothelial cell-pericyte axis in the development of novel therapeutic strategies.

The prostaglandin D2 antagonist asapiprant ameliorates clinical severity in young hosts infected with invasive Streptococcus pneumoniae.

Streptococcus pneumoniae (pneumococcus) remains a serious cause of pulmonary and systemic infections globally, and host-directed therapies are lacking. The aim of this study was to test the therapeutic efficacy of asapiprant, an inhibitor of prostaglandin D2 signaling, against pneumococcal infection. Treatment of young mice with asapiprant after pulmonary infection with invasive pneumococci significantly reduced systemic spread, disease severity, and host death. Protection was specific against bacterial dissemination from the lung to the blood but had no effect on pulmonary bacterial burden. Asapiprant-treated mice had enhanced antimicrobial activity in circulating neutrophils, elevated levels of reactive oxygen species (ROS) in lung macrophages/monocytes, and improved pulmonary barrier integrity indicated by significantly reduced diffusion of fluorescein isothiocyanate (FITC)-dextran from lungs into the circulation. These findings suggest that asapiprant protects the host against pneumococcal dissemination by enhancing the antimicrobial activity of immune cells and maintaining epithelial/endothelial barrier integrity in the lungs.

Quantification of Lymphangiogenesis in the Murine Lymphedema Tail Model Using Intravital Microscopy.

Background: Lymphedema is chronic limb swelling resulting from lymphatic dysfunction. It affects an estimated five million Americans. There is no cure for this disease. Assessing lymphatic growth is essential in developing novel therapeutics. Intravital microscopy (IVM) is a powerful imaging tool for investigating various biological processes in live animals. Tissue nanotransfection technology (TNT) facilitates a direct, transcutaneous nonviral vector gene delivery using a chip with nanochannel poration in a rapid (<100 ms) focused electric field. TNT was used in this study to deliver the genetic cargo in the murine tail lymphedema to assess the lymphangiogenesis. The purpose of this study is to experimentally evaluate the applicability of IVM to visualize and quantify lymphatics in the live mice model. Methods and Results: The murine tail model of lymphedema was utilized. TNT was applied to the murine tail (day 0) directly at the surgical site with genetic cargo loaded into the TNT reservoir: TNTpCMV6 group receives pCMV6 (expression vector backbone alone) (n = 6); TNTProx1 group receives pCMV6-Prox1 (n = 6). Lymphatic vessels (fluorescein isothiocyanate [FITC]-dextran stained) and lymphatic branch points (indicating lymphangiogenesis) were analyzed with the confocal/multiphoton microscope. The experimental group TNTProx1 exhibited reduced postsurgical tail lymphedema and increased lymphatic distribution compared to TNTpCMV6 group. More lymphatic branching points (>3-fold) were observed at the TNT site in TNTProx1 group. Conclusions: This study demonstrates a novel, powerful imaging tool for investigating lymphatic vessels in live murine tail model of lymphedema. IVM can be utilized for functional assessment of lymphatics and visualization of lymphangiogenesis following gene-based therapy.

The immune response to systemically administered endotoxin in the murine intestinal microcirculation under pentobarbital versus isoflurane anesthesia.

Pentobarbital and isoflurane are commonly used veterinary anesthetics. Due to the dangers of overdose by repeat-bolus regimen of pentobarbital, isoflurane has been recommended. However, literature suggests isoflurane-induced inhibition of cytokine and adhesion molecule release, impacting leukocyte adhesion. This study aims to characterize the impacts of pentobarbital versus isoflurane on leukocyte interactions within the intestinal microcirculation with and without endotoxin challenge. Female BALB/c mice were subjected to pentobarbital or isoflurane (N = 20) and challenged with endotoxin or saline by intraperitoneal injection. The mice were kept under anesthesia for 2 hours. Fluorochromes, rhodamine-6 G and fluorescein isothiocyanate, were injected intravenously. To visualize leukocyte adhesion within the intestinal microcirculation, laparotomy and intravital microscopy was performed. Leukocyte rolling and adhesion was quantified offline in a blinded fashion. Within collecting venules, leukocyte rolling and adhesion showed no significant differences between pentobarbital and isoflurane anesthesia under basal conditions. Endotoxin challenge caused a similar response in both anesthetic groups. Within postcapillary venules, no statistical differences between the two anesthetics were found for adhering leukocytes under basal conditions or following endotoxin challenge either. However, leukocyte rolling after LPS-challenge was significantly decreased in postcapillary venules during isoflurane anesthesia compared to pentobarbital anesthesia. Isoflurane anesthesia showed only minor differences in the immune response to endotoxin within the intestinal microcirculation compared to pentobarbital anesthesia. Due to the superior safety profile of volatile anesthetics, immunological studies may choose isoflurane over pentobarbital as the veterinary anesthetic of choice.

Adsorption study of eosin yellow and fluorescein dyes from aqueous solution using chitosan schiff base/polyvinyl acetate and its Fe2O3 nanocomposite

Adsorption study of eosin yellow and fluorescein dyes from aqueous solution using chitosan schiff base/polyvinyl acetate and its Fe2O3 nanocomposite

Activated carbon cellulose acetate based as effective adsorbent of contaminants from water

Various methods can be employed to remove pollutants from wastewater. Among these, adsorption using activated carbon materials has proven effective. This work proposes the use of a hazardous waste made from a semisynthetic polymer, cellulose acetate from cigarette butts, as raw material to produce activated carbons (AC). Four adsorbents were prepared from dirty and clean cigarette butts; two chemically activated carbons using phosphoric acid and two physically activated carbons using water vapor at 623 K. The carbons were characterized using FTIR, SEM, Nitrogen adsorption, and their points of zero charge. AC produced showed high potential as adsorbents for cleaning hazardous dyes from aqueous solutions. Adsorption isotherms were constructed for the textile dyes on the carbons, and these data were treated using Langmuir's equation to quantitatively describe the adsorption process. The adsorption of dyes with an initial concentration of 300 mg dm−3 showed adsorption percentages closed to the 100% for cationic dyes, while these oscillated between 40% and 100% for anionic dyes with the four activated carbons. The highest values of monolayer capacity (am) were achieved with the activated carbon prepared from cellulose acetate without prior treatment. Monolayer capacity with values ranging from 185 to 277 mg g−1 for cationic dyes and 126–178 mg g−1 for anionic dyes were obtained. Therefore, any waste of cellulose acetate can be used to obtain activated carbons with excellent adsorption capacities for dyes Methylene Blue, Crystal Violet, Malachite Green, Sirius Turquoise and Fluorescein.

Introduction of Organic Dyes into Cadmium(II) MOF for Tunable Light Emission and Highly Sensitive Probing Antibiotics

A honeycomb porous cadmium(II) metal–organic framework {[Cd(L)(H2O)]·NMP·5H2O}n (1, H2L = 1‐(4‐carboxyphenyl)‐1H‐pyrazole‐3‐carboxylic acid and NMP = 1‐methyl‐2‐pyrrolidinone) was obtained using the solvothermal method. The luminescent molecules rhodamine B (RhB) and fluorescein (FL) were encapsulated in 1 using the immersion method to prepare RhB@1 and FL@1 dual emission composites. Additionally, a series of RhBx/FL1‐x@1 composites with varying luminescence properties were generated by altering the molar ratio of RhB to FL. Furthermore, the photoluminescence of RhB@1 and FL@1 was demonstrated to be caused by photoelectron transfer and fluorescence resonance energy transfer from 1 to the encapsulated guest module. Notably, RhB@1 showed the considerable improvement in fluorescence selectivity and detection limit (10‐6 M) for metronidazole and dimetronidazole with remarkable recyclability.

Abstract PO4-27-10: A Systematic Review and Meta-analysis of Diagnostic Performance of Fluorescein Guided Sentinel Lymph Node Biopsy in Early Breast Cancer

Abstract Background: Evaluation of axillary lymph nodes status in cN0 axilla is performed by sentinel lymph node biopsy (SLNB) utilizing a combination of radioactive isotope and blue dye. There is a paucity of radioactive tracer in many countries of transitional economies. Hence, alternatives to isotope have been explored, viz. indocyanine green (ICG), superparamagnetic iron oxide nanoparticles and Fluorescein Sodium (FS). This systematic review and meta-analysis, evaluates the diagnostic performance of Fluorescein Sodium guided SLNB. Objectives: The objective was to evaluate the diagnostic performance of Fluorescein Sodium for sentinel lymph node biopsy in early breast cancer. Methods: PRISMA 2020 guidelines were followed for this meta-analysis. Eligibility criteria: Studies where SLNB was performed using Fluorescein Sodium for breast carcinoma. Information sources: PubMed, EMBASE, Cochrane library and online clinical trial registers Risk of bias: Articles were assessed for risk of bias using the QUADAS-2 tool. Synthesis of results: The main summary measures were pooled detection rate and pooled false negative rate using random effects model. In order to describe heterogeneity, we constructed summary receiver operating characteristic (SROC) curves with 95% prediction regions, estimated using bivariate meta-analysis with a test level random effect only, and forest plots. Calculations were performed using the Meta-Disc v1.4 program (available from https://metadisc.software.informer.com/1.4/). Results: Out of 45 studies found on initial systematic search, 7 studies were found eligible and were included in the meta-analysis based on data on 332 patients. The pooled detection rate was 93.2% (95% confidence interval [CI] 87% to 97%).). The heterogeneity (I2 value) of the pooled detection rate was 71%. Five validation studies were included for pooling the false negative rate (FNR) and included a total of 211 patients. The pooled FNR was 5.6% (95% CI; 2.9 to 9.07). Five validation studies were included for sensitivity, specificity, diagnostic odds ratio, LR+ and LR- meta-analysis. With regards to the identification of SLN the pooled sensitivity was 80% [(95% CI, 68% to 89%); I2=79.8]; specificity was 98% [(95% CI, 95% to 100%); I2=0.0]. The likelihood ratio for a positive test LR+ was 23.2 [(95% CI, 5.91 to 95.18); I2=0.0] and likelihood ratio for a negative test a LR- of 0.41 [(95% CI, 0.16 to 1.05); I2=88.1]. The diagnostic Odds ratio OR was 76.94 (95% CI, 15.88 to 372.79). Diagnostic accuracy as measured by SROC was 99.08%. Q index was 0.96. Publication bias for pooled detection rate as per Egger’s test was -2.2 (p= 0.66) and for FNR was 1.85 (p=0.29). Visual inspection of summary ROC curve and of forest plots did not show significant heterogeneity. Conclusion: Fluorescein guided SLNB is an affordable option for detection of lymph node metastases in cN0 patients with early breast cancer. It achieves a high detection rate of 93% with a false negative rate of 5.6% for the detection of axillary lymph node metastasis. We recommend larger multinational trials to confirm its high detection rate with low false negativity. Citation Format: Sanjay Kumar Yadav, Bharath S, Sarada Khadka, Dhananjaya Sharma, Chandan Kumar Jha, Gaurav Agarwal, Manju Singh, Anurag Srivastava, Amit Goyal. A Systematic Review and Meta-analysis of Diagnostic Performance of Fluorescein Guided Sentinel Lymph Node Biopsy in Early Breast Cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO4-27-10.

Fucoidan Improves Early Stage Diabetic Nephropathy via the Gut Microbiota-Mitochondria Axis in High-Fat Diet-Induced Diabetic Mice.

Diabetic nephropathy (DN) is a common microvascular complication of diabetes. Fucoidan, a polysaccharide containing fucose and sulfate group, ameliorates DN. However, the underlying mechanism has not been fully understood. This study aimed to explore the effects and mechanism of fucoidan on DN in high-fat diet-induced diabetic mice. A total of 90 C57BL/6J mice were randomly assigned to six groups (n = 15) as follows: normal control (NC), diabetes mellitus (DM), metformin (MTF), low-dose fucoidan (LFC), medium-dose fucoidan (MFC), and high-dose fucoidan (HFC). A technique based on fluorescein isothiocyanate (FITC-sinistin) elimination kinetics measured percutaneously was applied to determine the glomerular filtration rate (GFR). After 24 weeks, the mice were sacrificed and an early stage DN model was confirmed by GFR hyperfiltration, elevated urinary creatinine, normal urinary albumin, tubulointerstitial fibrosis, and glomerular hypertrophy. Fucoidan significantly improved the GFR hyperfiltration and renal fibrosis. An enriched SCFAs-producing bacteria and increased acetic concentration in cecum contents were found in fucoidan groups, as well as increased renal ATP levels and improved mitochondrial dysfunction. The renal inflammation and fibrosis were ameliorated through inhibiting the MAPKs pathway. In conclusion, fucoidan improved early stage DN targeting the microbiota-mitochondria axis by ameliorating mitochondrial oxidative stress and inhibiting the MAPKs pathway.

A novel integrated lateral flow immunoassay platform for the detection of cardiac troponin I using hierarchical dendritic copper-nickel nanostructures

Cardiac troponin I (cTnI) is a critical biomarker for the diagnosis of acute myocardial infarction (AMI). Herein, we report a novel integrated lateral flow immunoassay (LFIA) platform for highly sensitive point-of-care testing (POCT) of cTnI using hierarchical dendritic copper-nickel (HD-nanoCu-Ni) nanostructures. The electrodeposited HD-nanoCu-Ni film (∼22 μm thick) on an indium tin oxide (ITO) glass substrate exhibits superior capillary action and structural integrity. These properties enable efficient sample transport and antibody immobilization, making it a compelling alternative to conventional multi-component paper-based LFIA test strips, which are often plagued by structural fragility and susceptibility to moisture damage. The biofunctionalized HD-nanoCu-Ni substrates were laser-etched with lateral flow channels, including a sample loading/conjugate release zone, a test zone, and a control zone. Numerical simulations were used to further optimize the design of these channels to achieve optimal fluid flow and target capture. The HD-nanoCu-Ni LFIA device utilizes a fluorescence quenching based sandwich immunoassay format using antibody-labeled gold nanoparticles (AuNPs) as quenchers. Two different fluorescent materials, fluorescein isothiocyanate (FITC) and CdSe@ZnS quantum dots (QDs), were used as background fluorophores in the device. Upon the formation of a sandwich immunocomplex with cTnI on the HD-nanoCu-Ni device, introduced AuNPs led to the fluorescence quenching of the background fluorophores. The total assay time was approximately 15 min, demonstrating the rapid and efficient nature of the HD-nanoCu-Ni LFIA platform. For FITC, both inner filter effect (IFE) and fluorescence resonance energy transfer (FRET) contributed to the AuNP-mediated quenching. In the case of CdSe@ZnS QDs, IFE dominated the AuNP-induced quenching. Calibration curves were established based on the relationship between the fluorescence quenching intensity and cTnI concentration in human serum samples, ranging from 0.5 to 128 ng/mL. The limits of detection (LODs) were determined to be 0.27 ng/mL and 0.40 ng/mL for FITC and CdSe@ZnS QDs, respectively. A method comparison study using Passing-Bablok regression analysis on varying cTnI concentrations in human serum samples confirmed the equivalence of the HD-nanoCu-Ni LFIA platform to a commercial fluorescence cTnI LFIA assay kit, with no significant systematic or proportional bias observed.

Selective Timing of Maternal Immune Activation Alters Developmental Milestones, Behavior, and Blood-Brain Barrier Integrity

Background and Hypothesis: Maternal immune activation (MIA) has been associated with increased risk for neuropsychiatric disease. Fetal exposure to maternal infection activates different toll-like receptors (TLRs) to initiate innate inflammatory responses in mother and fetus. The goal of the current study is to determine sex-dependent aspects of MIA during mid-gestation on neurodevelopment. Methods: Timed-pregnant female mice were administered RQ (TLR7 agonist) or vehicle saline on embryonic day (E) 12.5. Maternal and fetal cytokines were measured to ensure infection of pregnant dam (E15). Offspring were assessed postnatally for developmental milestones and behavior. Adult offspring were perfused with fluorescein isothiocyanate (FITC; 100mg/mL) to visualize blood vessel integrity (FITC leakage; Frahm &amp; Tobet, 2015). To determine neural molecular mechanisms for behavioral changes, sections through the paraventricular nucleus of the hypothalamus (PVN) of offspring were immunolabeled for Glial Fibrillary Acidic Protein (GFAP; astrocytic end feet), Desmin (pericytes) and IBA-1 (microglia). Results: Maternal cytokines IL-6 (p&lt;0.05) and IL-10 (p&lt;0.01) were higher while TNFα (p&lt;0.01) and IL-17 were lower 3 days after RQ-exposure. In fetuses (E15), IL-6 (p&lt;0.05) and IL-17 (p&lt;0.0001) were greater, while IL-10 was higher only in males (p&lt;0.001) exposed to maternal infection. RQ-exposed males also had reduced TNFα (p&lt;0.05). Additionally, RQ-exposed offspring had lower body weights at birth and delayed eye openings. Females exposed to maternal RQ exhibited slower onset of puberty with delayed vaginal openings. On the behavioral side, juvenile and adult offspring of RQ injected moms exhibited less social-like behavior (p&lt;0.001 for all groups) with lower hedonic-like behavior selectively in females (p&lt;0.001). RQ-offspring further showed greater leakage in the PVN indicated by more FITC in the extravascular space (males p&lt;0.001, females p&lt;0.01). GFAP + (astrocytic end feet) coverage of FITC-labeled vessels was higher in the PVN selectively in RQ males (p&lt;0.001). Desmin+ (pericyte) coverage was greater in the PVN of RQ males (p&lt;0.01) and females (p&lt;0.001). Finally, there was greater number of IBA-1+ cells in the adult offspring PVN of both sexes (p&lt;0.01) after maternal injection of RQ. Conclusions: This study provides support for sex-dependent influences of fetal antecedents for altered brain development and behavioral outputs that could be indicative of increased susceptibility for adult disorders through immune mechanisms. Future studies will examine how timing of infection during gestation (mid vs. late) changes neurodevelopmental outputs in offspring. Supported by ORWH-NIMH U54 SCORE-MH118919. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Optical studies of fluorescein dye doped sulphamic acid single crystal

Fluorsceine, a dye used in lasers and non-linear optics have been studied for its interaction with crystal matrix (Sulphamic acid). Present study reveals the effect of fluorescein dye on sulphamic acid single crystals that are grown by slow evaporation solution technique at room temperature. Cell Parameters and crystal structure of pure sulphamic and 0.1 mol % fluorescein dye doped crystals is confirmed by Single Crystal XRD. The crystal structure is found to be orthorhombic and space group Pbca. Slight increase is noticed in cell parameters. Vibrational bands are assigned by FTIR and Raman spectroscopy. UV–Vis spectroscopy is also studied to find out band gap energy which is found to increase from 5.13 eV to 5.2 eV due to dye doping in crystal. Decrease in cutoff wavelength confirms that the synthesized crystals are permissible material for electro-optic application.