Humanization of a mouse anti-human complement C6 monoclonal antibody as a potential therapeutic for certain complement-mediated diseases.

The assembly of tissue-damaging membrane attack complexes (MACs; C5b-9) is a major mechanism by which excessive complement activation causes diseases. We previously developed a mouse anti-human C6 monoclonal antibody (mAb) 1C9 that selectively inhibits the assembly of MACs in human and non-human primates. In this project, we found that 1C9 also cross-reacted with rat and guinea pig C6, and determined its binding domains on C6 using different truncated C6 proteins. We then humanized the anti-C6 mAb by molecular modeling and complementarity-determining region grafting. After screening a library of 276 humanized variants with different combinations of humanized light and heavy chains in biophysical assays, we identified clone 3713 with the best developability profile, and an increased affinity against C6 when compared with the parental 1C9 mAb. This humanized 3713 mAb inhibited human, monkey, and rat complement-mediated hemolysis in vitro, and more importantly, it significantly reduced complement-mediated hemolysis in vivo in rats. These results demonstrated the successful humanization of the anti-C6 mAb and suggested that the humanized 3713 mAb could be further developed as a new therapeutic that selectively targets MAC for certain complement-mediated pathological conditions.

Development of a C3 Humanized Rat as a New Model for Evaluating Novel C3 Inhibitors.

INTRODUCTION: C3 is central for all complement activation pathways, thus making it an attractive therapeutic target. Many C3-targeted agents are under extensive development with one already approved for clinical use. However, most, if not all, C3 inhibitors are human or nonhuman primate C3-specific, making evaluating their efficacies in vivo before a clinical trial extremely difficult and costly. METHODS: We first studied the compatibility of human C3 in the rat complement system, then developed a C3 humanized rat using the CRISPR/Cas9 technology. We thoroughly characterized the resultant human C3 humanized rats and tested the treatment efficacy of an established primate-specific C3 inhibitor in a model of complement-mediated hemolysis in the C3 humanized rats. RESULTS: We found that supplementing human C3 protein into the C3-deficient rat blood restored its complement activity, which was inhibited by rat factor H or compstatin, suggesting that human C3 is compatible to the rat complement system. The newly developed C3 humanized rats appeared healthy and expressed human but not rat C3 without detectable spontaneous C3 activation. More importantly, complement-mediated hemolysis in the C3 humanized rats was also inhibited by compstatin both in vitro and in vivo. CONCLUSION: The successfully developed C3 humanized rats provided a much-desired rodent model to evaluate novel C3 inhibitors in vivo as potential drugs.

Effect of dissolved ozone flotation thickening process on coliform bacteria and antibiotics simultaneous abatement: A pilot-scale study.

This study focused on the removal of the total coliforms, fecal coliforms and four target antibiotics in the dissolved ozone flotation (DOF) thickening sludge process. Additionally, the thickened effluent chromaticity and its effect on thickened sludge hydrolysis process were investigated. Ozonation in the DOF process could inactivate coliforms by oxidizing cellular components and destroying genetic material, as well as altering the chemical structure of antibiotics, leading to the degradation of antibiotics. At an O3 dosage of 16 mg/g TS, the concentration of total coliforms and fecal coliforms decreased by 2.2 log and 2.4 log, corresponding to an overall removal rate of 99.4 % and 99.7 %, respectively. The total degradation rate of four target antibiotics (tetracycline (TC), oxytetracycline (OTC), norfloxacin (NOR), ofloxacin (OFL)) were 66.5 %, 68.8 %, 53.3 % and 57.5 %, respectively. The chromaticity removal rate of the thickened effluent reached 95 %. Analysis of fluorescence spectra indicated alterations in the fluorescence properties of dissolved organic matter, resulting in a decrease in fluorescence intensity by ozonation. The thickened sludge had higher hydrolysis rates, resulting in a greater production of volatile fatty acids (VFAs). This was mainly attributed to the increased amount of soluble protein and carbohydrate in the substrate after DOF treatment, which was more conducive for the rapid conversion of hydrolysis into VFAs during the initial stage. These results provided new ideas for upgrading and transforming the thickening process of wastewater treatment plants (WWTPs).

A CD6-targeted antibody-drug conjugate as a potential therapy for T cell-mediated disorders.

The selective targeting of pathogenic T cells is a holy grail in the development of new therapeutics for T cell-mediated disorders, including many autoimmune diseases and graft versus host disease. We describe the development of a CD6-targeted antibody-drug conjugate (CD6-ADC) by conjugating an inactive form of monomethyl auristatin E (MMAE), a potent mitotic toxin, onto a mAb against CD6, an established T cell surface marker. Even though CD6 is present on all T cells, only the activated (pathogenic) T cells vigorously divide and thus are susceptible to the antimitotic MMAE-mediated killing via the CD6-ADC. We found CD6-ADC selectively killed activated proliferating human T cells and antigen-specific mouse T cells in vitro. Furthermore, in vivo, whereas the CD6-ADC had no significant detrimental effect on normal T cells in naive CD6-humanized mice, the same dose of CD6-ADC, but not the controls, efficiently treated 2 preclinical models of autoimmune uveitis and a model of graft versus host disease. These results provide evidence suggesting that CD6-ADC could be further developed as a potential therapeutic agent for the selective elimination of pathogenic T cells and treatment of many T cell-mediated disorders.

Complement Receptor C3aR1 Contributes to Paclitaxel-Induced Peripheral Neuropathic Pain in Mice and Rats.

Cancer chemotherapy-induced neuropathic pain is a devastating pain syndrome without effective therapies. We previously reported that rats deficient in complement C3, the central component of complement activation cascade, showed a reduced degree of paclitaxel-induced mechanical allodynia (PIMA), suggesting that complement is integrally involved in the pathogenesis of this model. However, the underlying mechanism was unclear. Complement activation leads to the production of C3a, which mediates inflammation through its receptor C3aR1. In this article, we report that the administration of paclitaxel induced a significantly higher expression level of C3aR1 on dorsal root ganglion (DRG) macrophages and expansion of these macrophages in DRGs in wild-type (WT) compared with in C3aR1 knockout (KO) mice. We also found that paclitaxel induced less severe PIMA, along with a reduced DRG expression of transient receptor potential channels of the vanilloid subtype 4 (TRPV4), an essential mediator for PIMA, in C3aR1 KO than in WT mice. Treating WT mice or rats with a C3aR1 antagonist markedly attenuated PIMA in association with downregulated DRG TRPV4 expression, reduced DRG macrophages expansion, suppressed DRG neuron hyperexcitability, and alleviated peripheral intraepidermal nerve fiber loss. Administration of C3aR1 antagonist to TRPV4 KO mice further protected them from PIMA. These results suggest that complement regulates PIMA development through C3aR1 to upregulate TRPV4 on DRG neurons and promote DRG macrophage expansion. Targeting C3aR1 could be a novel therapeutic approach to alleviate this debilitating pain syndrome.

D-2-hydroxyglutarate regulates human brain vascular endothelial cell proliferation and barrier function.

Gain-of-function mutations in isocitrate dehydrogenase (IDH) genes result in excessive production of (D)-2-hydroxyglutarate (D-2HG) which intrinsically modifies tumor cell epigenetics and impacts surrounding noncancerous cells through nonepigenetic pathways. However, whether D-2HG has a paracrine effect on endothelial cells in the tumor microenvironment needs further clarification. We quantified microvessel density by immunohistochemistry using tissue sections from 60 high-grade astrocytic gliomas with or without IDH mutation. Microvessel density was found to be reduced in tumors carrying an IDH mutation. Ex vivo experiments showed that D-2HG inhibited endothelial cell migration, wound healing, and tube formation by suppressing cell proliferation but not viability, possibly through reduced activation of the mTOR/STAT3 pathway. Further, D-2HG reduced fluorescent dextran permeability and decreased paracellular T-cell transendothelial migration by augmenting expression of junctional proteins thereby collectively increasing endothelial barrier function. These results indicate that D-2HG may influence the tumor vascular microenvironment by reducing the intratumoral vasculature density and by inhibiting the transport of metabolites and extravasation of circulating cells into the astrocytoma microenvironment. These observations provide a rationale for combining IDH inhibition with antitumor immunological/angiogenic approaches and suggest a molecular basis for resistance to antiangiogenic drugs in patients whose tumors express a mutant IDH allele.

CD6-targeted antibody-drug conjugate as a new therapeutic agent for T cell lymphoma.

T cell lymphomas (TCL) are heterogeneous, aggressive, and have few available targeted therapeutics. In this study, we determined that CD6, an established T cell marker, was expressed at high levels on almost all examined TCL patient specimens, suggesting that CD6 could be a new therapeutic target for this life-threatening blood cancer. We prepared a CD6-targeted antibody-drug conjugate (CD6-ADC) by conjugating monomethyl auristatin E (MMAE), an FDA-approved mitotic toxin, to a high-affinity anti-human CD6 monoclonal antibody (mAb). In contrast to both the unconjugated anti-CD6 mAb, and the non-binding control ADC, CD6-ADC potently and selectively killed TCL cells in vitro in both time- and concentration-dependent manners. It also prevented the development of tumors in vivo in a preclinical model of TCL. More importantly, systemic or local administration of the CD6-ADC or its humanized version, but not the controls, significantly shrank established tumors in the preclinical mouse model of TCL. These results suggest that CD6 is a novel therapeutic target in TCLs and provide a strong rationale for the further development of CD6-ADC as a promising therapy for patients with these potentially fatal lymphoid neoplasms.

Mechano-Driven Tribo-Electrophoresis Enabled Human-Droplet Interaction in 3D Space.

Electronically controlled droplet manipulation has widespread applications in biochemistry, life sciences, and industry. However, current technologies such as electrowetting, electrostatics, and surface charge printing rely on complex electrode arrays and external power supplies, leading to inefficient manipulation. In light of these limitations, a novel method is proposed, which leverages tribo-electrophoresis (TEP) to pipette in an oil medium, thereby enabling human-droplet interactions to be constructed with greater efficiency. The approach involves the rational design of a triboelectric nanogenerator-electrostatic tweezer that generates an electric field to charge the droplet and improves the maneuverability of the charged droplet, including aligned/non-aligned pipetting and stable transport in the clamped state, which can be accomplished solely by hand motion. The TEP method not only provides droplets with freedom to move in three dimensions but also offers a feasibility case for chemical reactions in the liquid phase and non-invasive sample extraction. This breakthrough establishes a cornerstone for human-droplet interactions capitalized on triboelectric nanogenerators, opening new avenues for research in droplet manipulation.

A nanobody-based complement inhibitor targeting complement component 2 reduces hemolysis in a complement humanized mouse model of autoimmune hemolytic anemia.

C2 is an attractive therapeutic target for many complement-mediated diseases. We developed Nab1B10, a new anti-C2 nanobody that potently and selectively inhibits both the classical and lectin pathways of complement activation. Mechanistically, Nab1B10 binds to the C2a portion of C2 and inhibits the assembly of C3 convertase C4b2a. Nab1B10 cross-reacts with monkey but not rodent C2 and inhibits classical pathway-mediated hemolysis. Using a new complement humanized mouse model of autoimmune hemolytic anemia (AIHA), we demonstrated that Nab1B10 abolished classical pathway complement activation-mediated hemolysis in vivo. We also developed C2-neutralizing bi- and tetra-valent antibodies based on Nab1B10 and found these antibodies significantly more potent than the other anti-C2 monoclonal antibody that is already in clinical trials. These data suggest that these novel C2-neutralizing nanobodies could be further developed as new therapeutics for many complement-mediated diseases, in which pathogenesis is dependent on the classical and/or lectin pathway of complement activation.

Plasmon-driven catalytic reactions in optoplasmonic sandwich hybrid structure.

As an interesting phenomenon in the field of surface enhanced Raman spectroscopy (SERS), the plasmon-driven catalytic reaction (PDSC) induced by plasmonic hot electrons has great value in the research of novel properties of surface plasmons and accuracy of SERS applications. In this work, an optoplasmonic sandwich hybrid structure is proposed for studying PDSC of p-aminothiophenol (PATP) molecules, which is composed of Au film, metal organic frameworks (MOFs) nanoparticles, zeolithic imidazolate (ZIF-8), and single S i O 2 microsphere (Au f i l m@M O F s@S i O 2). In order to analyze the novel, to the best of our knowledge, phenomenon of the PDSC in this micro-nano structure, the hot electron generation in the MOF without the plasmonic core is carried out by combining the plasmonic enhancement of gold film with the light concentration of microspheres. Experimental data show that the PDSC reactions is dependent on the size of the MOFs nanoparticle and the size of the S i O 2 microsphere, which is confirmed by the electromagnetic field simulation of the finite-difference time-domain method (FDTD). Our work not only strengthens the understanding of surface plasmon in optoplasmonic hybrid structures but also has broad application prospects in the SERS and plasmon-driven catalytic fields.

Mental health status of infected children between 7 to 12 years old in Fangcang Shelter Hospital during the COVID-19 Shanghai lockdown in 2022: a cross-section study.

Background: There has been an increase in research on the potential adverse effects on children's mental health, especially depression and anxiety, during the coronavirus disease 2019 (COVID-19) pandemic over the past few months. Therefore, the aim of the present study was to investigate depression and anxiety symptoms among children in shelter hospitals during the 2022 Shanghai lockdown. Methods: A total of 98 infected children aged 7-12 years were enrolled in this study between April 19 and June 4, 2022. The Children's Depression Inventory (CDI), Anxiety Scale for Children-Autism Spectrum Disorder (ASC-ASD), and Anxiety Scale or Children-Autism Spectrum Disorder Parent Form (ASC-ADS-P) were used to assess children's depression and anxiety symptoms. Children's guardians completed the survey by verbally asking their child/children the questions. The guardians additionally completed the ASC-ASD-P. Results: The prevalence of depression and anxiety was 12.2% and 13.3%, respectively. A total of 66 respondents reported no physical symptoms. Linear regression showed that myalgia [7.198, 95% confidence interval (CI): 3.163-11.232], headache (7.189, 95% CI: 3.842-10.535) coryza (5.362, 95% CI: 2.654-8.070), and number of quarantine days (4.378, 95% CI: 3.409-5.348) were significantly correlated with higher levels of depression, whereas chills (14.337, 95% CI: 9.799-18.875), coryza (9.309, 95% CI: 6.467-12.152), headache (7.193, 95% CI: 3.182-11.204), myalgia (5.571, 95% CI: 0.684-10.459), number of quarantine days (3.190, 95% CI: 1.796-4.584), and gender (male) (-4.137, 95% CI: -6.609 to 1.665) were associated with anxiety scores. Persistent fever was correlated with depression (P=0.007), whereas physical discomfort, such as persistent fever, cough, sore throat, headache, myalgia, and coryza were correlated with anxiety (all P<0.05). Conclusions: The findings of the present study indicated a moderate prevalence of depression and anxiety among infected children in a shelter hospital during the 2022 Shanghai lockdown. Therefore, the findings of this study could provide scientific basis for the development of targeted psychological intervention. It could be helpful for policy-makers to focus on psychological health among infected children and help to optimize future interventions.

Inhibitory effect of ficin on Candida albicans biofilm formation and pre-formed biofilms.

BACKGROUND: To investigate the effect of ficin, a type of proteases, on Candida albicans (C. albicans) biofilm, including forming and pre-formed biofilms. METHODS: Crystal violet tests together with colony forming unit (CFU) counts were used to detect fungal biofilm biomass. Live/dead staining of biofilms observed by confocal laser scanning microscopy was used to monitor fungal activity. Finally, gene expression of C. albicans within biofilms was assessed by qRT-PCR. RESULTS: According to our results, biofilm biomass was dramatically reduced by ficin in both biofilm formation and pre-formed biofilms, as revealed by the crystal violet assay and CFU count (p < 0.05). Fungal activity in biofilm formation and pre-formed biofilms was not significantly influenced by ficin according to live/dead staining. Fungal polymorphism and biofilm associated gene expression were influenced by ficin, especially in groups with prominent antibiofilm effects. CONCLUSIONS: In summary, ficin effectively inhibited C. albicans biofilm formation and detached its preformed biofilm, and it might be used to treat C. albicans biofilm associated problems.

CDCP1 regulates retinal pigmented epithelial barrier integrity for the development of experimental autoimmune uveitis.

Cub domain-containing protein 1 (CDCP1) is a protein that is highly expressed on the surface of many cancer cells. However, its distribution in normal tissues and its potential roles in nontumor cells are poorly understood. We found that CDCP1 is present on both human and mouse retinal pigment epithelial (RPE) cells. CDCP1-KO mice developed attenuated retinal inflammation in a passive model of autoimmune uveitis, with disrupted tight junctions and infiltrating T cells detected in RPE flat mounts from WT but not CDCP1-KO mice during EAU development. Mechanistically, we discovered that CDCP1 on RPE cells was upregulated by IFN-γ in vitro and after EAU induction in vivo. CD6 stimulation induced increased RPE barrier permeability of WT but not CDCP1-knockdown (CDCP1-KD) RPE cells, and activated T cells migrated through WT RPE monolayers more efficiently than the CDCP1-KD RPE monolayers. In addition, CD6 stimulation of WT but not the CDCP1-KD RPE cells induced massive stress fiber formation and focal adhesion disruption to reduce cell barrier tight junctions. These data suggest that CDCP1 on RPE cells interacts with CD6 on T cells to induce RPE cytoskeleton remodeling and focal adhesion disruption, which open up the tight junctions to facilitate T cell infiltration for the development of uveitis.

A novel application of dissolved ozone flotation on sewage sludge thickening: Performance and mechanism investigation.

Sludge thickening in sewage treatment plants is an essential step to reduce the sewage sludge volume and provide space for collaborative anaerobic digestion of sludge and other urban organic wastes. Dissolved ozone flotation (DOF) is a novel choice worthy of application in the field of sludge thickening. In order to investigate the effect of DOF thickening, the total solid content (TS %) was used to characterize the thickening performance under different O3 dosage. The optimal condition was determined to be polyacrylamide (PAM) dosage = 3 ‰ TS, air floatation time = 2 h and O3 dosage = 12 mg/g TS, under which the TS % of raw-sewage sludge (RS) increased from 0.33 ± 0.01 % to 8.03 ± 0.06 %. In this study, the relationship between the sludge thickening performance, physicochemical properties, and the changes of organic matter (content, structure and molecular weight) in extracellular polymers (EPS) was systematically clarified. The results indicated that the DOF couple with PAM could change the sludge surface properties, destroy the sludge floc structure, release intracellular organic matter, and increase moisture fluidity. The surface hydrophilicity/hydrophobicity, protein (PN) secondary structure and moisture distribution were mainly responsible for sludge thickening performance. Moreover, the change of TS % during the DOF thickening process was mainly caused by the variations of the organic matter content in EMPS layer. The identification of key influencing factors was conducive to the further regulation and upgrading of the novel application for enhanced sludge thickening in sewage treatment plants.

Optoplasmonic MOFs film for SERS detection.

Optoplasmonic hybrid structures composed of photonic and plasmonic elements with excellent optical properties are of great significance for the development of surface-enhanced Raman spectroscopy (SERS) substrates. In this work, the optoplasmonic hybrid structure is composed of SiO2 microsphere and two-dimensional (2D) plasmonic- metal organic frameworks (MOF) film. Among them, the 2D plasmonic-MOF film is prepared from silver nanoparticles encapsulated by zeolitic imidazole acid framework (AgNP@ZIF-8) by self-assembly method. This optoplasmonic hybrid structure with gas adsorption properties could be used as a SERS substrate for 4-Mercaptophenol (4-MP) gas detection. Experimental data show that this substrate is dependent on the thickness of the ZIF shell and the size of the SiO2 microspheres. In addition, it is confirmed by the electromagnetic field simulation of finite-difference time-domain method (FDTD). The optoplasmonic hybrid microstructures exhibit good uniformity for detection of 4-MP gas molecules. This work not only broadens the understanding of our optoplasmonic hybrid structure, but also has broad application prospects in SERS and gas sensing related fields.

The suppression effect of SCH-79797 on Streptococcus mutans biofilm formation.

Background: SCH-79797 was recently shown to be a broad-spectrum antibacterial agent with a dual-bactericidal mechanism. However, its anti-biofilm effect remains unknown. Purpose: To investigate the effect of SCH-79797 on the biofilm formation of the cariogenic Streptococcus mutans. Methods and Results: Crystal violet staining, colony forming units count and MTT assays (for cell metabolic activity) revealed that S. mutans biofilm formation was significantly suppressed. In addition, virulence factors, including extracellular polysaccharides (investigated by bacterial/exopolysaccharide staining and the anthrone method) and acid production (investigated by lactic acid and supernatant pH detection) were also inhibited significantly. Moreover, the biofilm inhibitory effect of SCH-79797 was mediated through its repression of bacterial growth and not by a bactericidal effect, which was verified by growth curve and bacterial live/ dead staining, respectively. Quantitative real-time PCR results disclosed that SCH-79797 affected bacterial acid production and tolerance, polysaccharide synthesis and remodeling, biofilm formation and quorum sensing-related gene expression. In addition, SCH-79797 showed good biocompatibility as determined by cytotoxicity assays. Conclusion: SCH-79797 had an anti-biofilm effect and showed application prospects in the control of dental caries.

ROS-Induced Oxidative Damage and Mitochondrial Dysfunction Mediated by Inhibition of SIRT3 in Cultured Cochlear Cells.

Sensorineural hearing loss (SNHL) is one of the most common causes of disability worldwide. Previous evidence suggests that reactive oxygen species (ROS) may play an important role in the occurrence and development of SNHL, while its mechanism remains unclear. We cultured dissected organs of Corti in medium containing different concentrations (0, 0.25, 0.5, 0.75, 1, and 1.25 mM) of hydrogen peroxide (H2O2) and established a four-concentration model of 0, 0.5, 0.75, and 1 mM to study different degrees of damage. We examined ROS-induced mitochondrial damage and the role of sirtuin 3 (SIRT3). Our results revealed that the number of ribbon synapses and hair cells appeared significantly concentration-dependent decrease with exposure to H2O2. Outer hair cells (OHCs) and inner hair cells (IHCs) began to be lost, and activation of apoptosis of hair cells (HCs) was observed at 0.75 mM and 1 mM H2O2, respectively. In contrast with the control group, the accumulation of ROS was significantly higher, and the mitochondrial membrane potential (MMP) was lower in the H2O2-treated groups. Furthermore, the expression of SIRT3, FOXO3A, and SOD2 proteins declined, except for an initial elevation of SIRT3 between 0 and 0.75 mM H2O2. Administration of the selective SIRT3 inhibitor 3-(1H-1,2,3-triazol-4-yl) pyridine resulted in increased damage to the cochlea, including loss of ribbon synapses and hair cells, apoptosis of hair cells, more production of ROS, and reduced mitochondrial membrane potential. Thoroughly, our results highlight that ROS-induced mitochondrial oxidative damage drives hair cell degeneration and apoptosis. Furthermore, SIRT3 is crucial for preserving mitochondrial function and protecting the cochlea from oxidative damage and may represent a possible therapeutic target for SNHL.

Rich Structural Index for Stereoscopic Image Quality Assessment.

The human visual system (HVS), affected by viewing distance when perceiving the stereo image information, is of great significance to study of stereoscopic image quality assessment. Many methods of stereoscopic image quality assessment do not have comprehensive consideration for human visual perception characteristics. In accordance with this, we propose a Rich Structural Index (RSI) for Stereoscopic Image objective Quality Assessment (SIQA) method based on multi-scale perception characteristics. To begin with, we put the stereo pair into the image pyramid based on Contrast Sensitivity Function (CSF) to obtain sensitive images of different resolution. Then, we obtain local Luminance and Structural Index (LSI) in a locally adaptive manner on gradient maps which consider the luminance masking and contrast masking. At the same time we use Singular Value Decomposition (SVD) to obtain the Sharpness and Intrinsic Structural Index (SISI) to effectively capture the changes introduced in the image (due to distortion). Meanwhile, considering the disparity edge structures, we use gradient cross-mapping algorithm to obtain Depth Texture Structural Index (DTSI). After that, we apply the standard deviation method for the above results to obtain contrast index of reference and distortion components. Finally, for the loss caused by the randomness of the parameters, we use Support Vector Machine Regression based on Genetic Algorithm (GA-SVR) training to obtain the final quality score. We conducted a comprehensive evaluation with state-of-the-art methods on four open databases. The experimental results show that the proposed method has stable performance and strong competitive advantage.

Regulatory Effect of Irresistin-16 on Competitive Dual-Species Biofilms Composed of Streptococcus mutans and Streptococcus sanguinis.

Based on the ecological plaque hypothesis, suppressing opportunistic pathogens within biofilms, rather than killing microbes indiscriminately, could be a biofilm control strategy for managing dental caries. The present study aimed to evaluate the effects of irresistin-16 (IRS-16) on competitive dual-species biofilms, which consisted of the conditional cariogenic agent Streptococcus mutans (S. mutans) and oral commensal bacteria Streptococcus sanguinis (S. sanguinis). Bacterial growth and biofilm formation were monitored using growth curve and crystal violet staining, respectively. The microbial proportion was determined using fluorescence in situ hybridization. A 2, 5-diphenyltetrazolium bromide assay was used to measure the metabolic activity of biofilms. Bacterial/extracellular polysaccharide (EPS) dyeing, together with water-insoluble EPS measurements, were used to estimate EPS synthesis. A lactic acid assay was performed to detect lactic acid generation in biofilms. The cytotoxicity of IRS-16 was evaluated in mouse fibroblast L929 cells using a live/dead cell viability assay and cell counting kit-8 assay. Our results showed that IRS-16 exhibited selective anti-biofilm activity, leading to a remarkable survival disadvantage of S. mutans within competitive dual-species biofilms. In addition, the metabolic activity, EPS synthesis, and acid generation of dual-species biofilms were significantly reduced by IRS-16. Moreover, IRS-16 showed minimal cytotoxicity against mouse fibroblast L929 cells. In conclusion, IRS-16 exhibited remarkable regulatory effects on dual-species biofilms composed of S. mutans and S. sanguinis with low cytotoxicity, suggesting that it may have potential for use in caries management through ecological biofilm control.

Hemorrhage Risk Profiles among Different Antithrombotic Regimens: Evidence from a Real-World Analysis of Postmarketing Surveillance Data.

BACKGROUND: Although the use of direct oral anticoagulants (DOACs) has been reported in patients with atrial fibrillation (AF), there is currently no consensus on the occurrence or characteristics of the hemorrhage risk in different antithrombotic regimens. METHODS: Disproportionality and Bayesian analyses were performed in mining data of suspected hemorrhagic events after antithrombotic drug use from the FDA Adverse Event Reporting System (FAERS) from January 2004 to September 2019. The time to onset and fatality rate of hemorrhage following different antithrombotic regimens were also compared. RESULTS: A total of 84,998 reports of hemorrhage-related adverse events with the use of antithrombotic drugs were identified. The patients included were mostly from the Americas (80.87%) and Europe (13.22%), with most data submitted by nonhealthcare professionals. Among the seven antithrombotic drug monotherapies, betrixaban had the highest association with hemorrhage based on the highest reporting odds ratio (ROR, 829.95; 95% CI = 113.61-6063.15), proportional reporting ratio (PRR, 24.68, χ2 = 804.24), and multi-item gamma Poisson shrinker (MGPS, 24.68, 95% one-sided CI = 4.67). The combination therapies of clopidogrel plus new oral anticoagulants had higher RORs, PRRs, and empirical Bayesian geometric means (EBGMs) than the antithrombotic drug monotherapies. Hemorrhage associated with rivaroxaban plus clopidogrel appeared to have an earlier onset (171 days vs 219 days, 95% two-sided CI =68.68-27.34, p < 0.0001) and a lower fatality rate (15.30% vs 17.74%, p<0.05) than that associated with rivaroxaban monotherapy. CONCLUSION: This study provides a relevant overview of the hemorrhagic complications/fatalities associated with different antithrombotic regimens in their real-world use. Among the combination therapies, clopidogrel plus DOACs were found to have stronger associations with hemorrhage than traditional dual antithrombotic therapies. Rivaroxaban showed a stronger association with hemorrhage than other antithrombotic drug monotherapies, and apixaban monotherapy appeared to have weaker associations with hemorrhage than others.