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Objective: Unknown etiology Background: This case report illustrates the incidental detection of atypical hyporeflective bleb-like structures in an otherwise asymptomatic human cornea, highlighting the effectiveness of non-invasive biophotonic imaging techniques such as optical coherence tomography (OCT) and large-area confocal laser scanning microscopy (CLSM). Case Report: A 57-year-old man underwent a comprehensive ophthalmological examination, including slit-lamp biomicroscopy, corneal topography, and densitometry, as part of a clinical study. The patient presented with a clear cornea, a deep and optically empty anterior chamber, and a clear crystalline lens in both eyes. Best-corrected distance visual acuity was -0.1 logMAR. He denied ocular pain, tearing, or photophobia. There was no history of ocular trauma, infectious or genetic diseases, or ocular surgery. Further, OCT and large-area CLSM were employed for a more in-depth analysis of the corneal structure. Standard ophthalmological assessments indicated normal ocular health with no apparent corneal abnormalities. High-resolution OCT and large-area CLSM imaging revealed atypical hyporeflective bleb-like structures within the corneal epithelium and sub-basal nerve plexus but not in the stroma. Conclusions: The authors hypothesize that the observed findings may indicate an early stage of epithelial basement membrane dystrophy, potentially preceding the manifestation of clinical symptoms, detectable solely through advanced biophotonic imaging methods. It is important to emphasize that these observations do not represent a definitive diagnosis. Nevertheless, the discovery of these atypical structures via advanced imaging underscores the importance of incorporating non-invasive biophotonic techniques into preoperative eye examinations in certain situations. This approach could significantly improve the early detection and management of corneal diseases, leading to improved patient outcomes. [ABSTRACT FROM AUTHOR]

Non‐invasive diagnostics like line‐field confocal optical coherence tomography (LC‐OCT) are being implemented in dermato‐oncology. However, unification of terminology in LC‐OCT is lacking. By reviewing the LC‐OCT literature in the field of dermato‐oncology, this study aimed to develop a unified terminological glossary integrated with traditional histopathology. A PRISMA‐guided literature‐search was conducted for English‐language publications on LC‐OCT of actinic keratosis (AK), keratinocyte carcinoma (KC), and malignant melanoma (MM). Study characteristics and terminology were compiled. To harmonize LC‐OCT terminology and integrate with histopathology, synonymous terms for image features of AK, KC, and MM were merged by two authors, organized by skin layer and lesion‐type. A subset of key LC‐OCT image‐markers with histopathological correlates that in combination were typical of AK, squamous cell carcinoma in situ (SCCis), invasive squamous cell carcinoma (SCC), basal cell carcinoma (BCC), and MM in traditional histopathology, were selected from the glossary by an experienced dermatopathologist. Seventeen observational studies of AK (7 studies), KC (13 studies), MM (7 studies) utilizing LC‐OCT were included, with 117 terms describing either AK, KC, or MM. These were merged to produce 45 merged‐terms (61.5% reduction); 5 assigned to the stratum corneum (SC), 23 to the viable epidermis, 2 to dermo‐epidermal junction (DEJ) and 15 to the dermis. For each lesion, mandatory key image‐markers were a well‐defined DEJ and presence of mild/moderate but not severe epidermal dysplasia for AK, severe epidermal dysplasia and well‐defined DEJ for SCCis, interrupted DEJ and/or dermal broad infiltrative strands for invasive SCC, dermal lobules connected and/or unconnected to the epidermis for BCC, as well as single atypical melanocytes and/or nest of atypical melanocytes in the epidermis or dermis for MM. This review compiles evidence on LC‐OCT in dermato‐oncology, providing a harmonized histopathology‐integrated terminology and key image‐markers for each lesion. Further evaluation is required to determine the clinical value of these findings. [ABSTRACT FROM AUTHOR]

Abstract Background: Currently, there is no uniform and official terminology in Portuguese for reflectance confocal microscopy analysis, despite the increasing number of Brazilian dermatologists using this new tool. Objective: To present the terminology in Brazilian Portuguese for the description of reflectance confocal microscopy and establish the first Brazilian consensus on terms related to normal skin and cutaneous tumors. Methods: 10 Brazilian specialists from different institutions and states of Brazil were selected to evaluate the best corresponding terms in Portuguese for normal skin, melanocytic and non-melanocytic tumors. The terms used were translated from international consensuses in the English language. The modified Delphi method was used to create the consensus in 3 steps. Results: The terms considered the most appropriate in the Portuguese language to describe the findings of normal skin, melanocytic and non-melanocytic lesions in the reflectance confocal microscopy analysis were presented. Study limitations: The limitations of the present study include the number of participants and limited regional representation (only two of the five Brazilian regions were represented). Conclusion: This Brazilian consensus represents an opportunity for dermatologists and physicians specializing in cutaneous oncology to become familiar with reflectance confocal microscopy, propagating the technique in clinical and research environments to stimulate national and international publications on this subject.

To describe a case of treatment-resistant corneal infiltrate in a patient 4 months after penetrating keratoplasty. The etiological diagnosis was established through microbiology and in vivo corneal confocal microscopy. A 80-year-old female patient presented with stromal infiltrate in the donor corneal bud 4 months following penetrating keratoplasty. Initially, the corneal culture was positive for Candida parapsilosis. In vivo corneal confocal microscopy demonstrated approximately 4 µm hyperreflective spherical microorganisms with a suggestive Candida spp. A second corneal culture was positive for Candida sp., and hyphae presence in the analyzed material was observed. The patient remained refractory to therapy. Confocal microscopy of the cornea continues to grow as a useful, non-invasive diagnostic method and in monitoring response to therapy. This report contributes by detecting Candida sp. in in vivo corneal confocal microscopy to aid in the diagnosis of keratitis. [ABSTRACT FROM AUTHOR]

Abstract Since its first introduction into medical practice, reflectance confocal microscopy (RCM) has been a valuable non-invasive diagnostic tool for the assessment of benign and malignant neoplasms of the skin. It has also been used as an adjunct for diagnosing equivocal cutaneous neoplasms that lack characteristic clinical or dermoscopic features. The use of RCM has led to a decreased number of biopsies of benign lesions. Multiple published studies show a strong correlation between RCM and histopathology thereby creating a bridge between clinical aspects, dermoscopy, and histopathology. Dermatopathologists may potentially play an important role in the interpretation of confocal images, by their ability to correlate histopathologic findings. RCM has also been shown to be an important adjunct to delineating tumoral margins during surgery, as well as for monitoring the non-surgical treatment of skin cancers. Advanced technology with smaller probes, such as the VivaScope 3000, has allowed access to lesions in previously inaccessible anatomic locations. This review explains the technical principles of RCM and describes the most common RCM features of normal skin with their corresponding histological correlation.

In vivo reflectance confocal microscopy (RCM) is poorly investigated in oral pathology due to the peculiar anatomical and topographical oral mucosa features. A dedicated handheld confocal microscope with an intra‐oral probe was developed for oral mucosa imaging. The main objective was to describe the healthy oral mucosa and the cytoarchitectural findings detectable in different oral disorders by means of the newly designed handheld confocal microscope. Secondary aim was to identify the main RCM criteria that differentiate oral lesions in order to provide algorithm for a rapid non‐invasive evaluation. This observational retrospective study included all consecutive patients with oral disorders and volunteers with healthy oral mucosa who underwent RCM examination in our outpatient clinic from September 2018 to December 2021. Three different investigators examined together the RCM images to detect the key features and secondary criteria for each type of oral lesion collected. The study population included 110 patients affected by oral lesions and seven volunteers with healthy oral mucosae. A total of 15 oral disorders were imaged and divided in three main groups: white, red and pigmented lesions. Key features and secondary criteria were identified for every single type of oral disease. RCM permits a cytoarchitectural evaluation of the oral mucosae affected by inflammatory, dysplastic and neoplastic diseases, thus orienting the clinicians towards non‐invasive diagnosis and enhancing the diagnostic management. The "tree diagrams" proposed allow a schematic and simplified view of confocal features for each type of oral disease, thus drastically reducing the diagnostic timing. [ABSTRACT FROM AUTHOR]

Purpose: Preclinical studies have confirmed the safety and efficacy of narrowband low-intensity ultraviolet C light (UVC) in managing bacterial corneal infection. To further consolidate these findings, the present study aimed to explore in vitro anti-biofilm efficacy of low-intensity UVC light for its potential use in biofilm-related infections., Methods: Pseudomonas aeruginosa biofilm was grown in chamber well slides for 48 h and exposed to one of the following challenges: UVC (265 nm wavelength, intensity 1.93 mW/cm 2 ) for 15 s, 30 s, 60 s or 120 s duration, 70% propanol (positive control), or no exposure (negative control). Bacterial LIVE/DEAD staining was conducted at 1 h, 4 h, 6 h and 8 h after challenge exposures to assess the temporal pattern of biofilm inactivation, and slides were imaged using confocal microscopy. Treatment efficacy was quantified by dead biofilm biomass (volume/area - μm 3 /μm 2 ) for different treatment groups at each time point., Results: At each time point post-exposure, dead biofilm biomass was consistently higher in the alcohol- and UVC-challenged groups than in the unchallenged control (p < 0.05), suggesting a sustained biocidal impact after a given challenge. The quantity of dead biofilm biomass did not differ between UVC groups at any time point (p > 0.05). Observed by confocal microscopy, UVC-exposed biofilm demonstrated predominantly intermediate-stage biofilm (i.e., dying state) at 1 h, which progressed to dead biofilm by 4 h., Conclusion: Low doses of UVC demonstrated potent anti-biofilm activity, even in exposures as short as 15 s, the dose that has previously been deemed to be effective in managing corneal infection in vivo. These data support the potential for this UVC light-based technology to serve as an affordable, convenient, and effective means of treating ocular infections associated with bacterial biofilm., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Simon J. Dean is a shareholder in Photon Therapeutics Limited, which holds intellectual property relating to the therapeutic use of UVC. The other authors have no commercial or proprietary interest in any concept or product described in this article. Jennifer P. Craig reports a spousal relationship with Simon J. Dean and financial support and equipment, drugs, or supplies provided by Photon Therapeutics Ltd. to the University of Auckland for investigator-initiated research. Simon J. Dean reports a relationship with Photon Therapeutics Ltd. that includes: board membership and equity or stocks. Simon J. Dean has patent ##WO2022013435A2·2022-01-20 pending to Photon Therapeutics Ltd. No other authors have any known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier B.V. All rights reserved.)

Quinone outside inhibitor (QoI) fungicide resistance in Alternaria alternata populations was reported in Brazil for the first time in 2019, in São Paulo orchards, and the mutation G143A in cytochrome b (cytb) was found in resistant isolates. Our study investigated the infectious process, production of reactive oxygen species (ROS), and fungal cell death in resistant (QoI-R) and sensitive (QoI-S) A. alternata pathotype tangerine (Aapt) isolates. Morphological characterization of Aapt isolates was performed using confocal laser scanning microscopy (CLSM). Alternaria brown spot (ABS) symptoms were produced by Aapt isolates on tangelo cv. BRS Piemonte. Germination of QoI-R conidia and production of germ tubes on tangelo leaflets treated with 100 μg mL -1 of pyraclostrobin 18 h after inoculation (hai) was observed using scanning electron microscopy (SEM). At the same time, QoI-S conidial germination was inhibited on tangelo leaflets treated with pyraclostrobin. ROS production and cell death in Aapt isolates at high fungicide concentrations were observed using CLSM. QoI-S conidia exhibited high ROS production, indicating high oxidative stress. When dyed with propidium iodate (PI), QoI-S conidia emitted red fluorescence, showing cell death and confirming their sensitive phenotype. In contrast, QoI-R conidia neither produced ROS nor exhibited red fluorescence, indicating no cell death and confirming their resistant phenotype. Therefore, our findings evidence that microscopic techniques may help characterize events during fungi-plant interactions, ROS production, cell death, and Aapt phenotypes resistant and sensitive to QoIs using fluorometric protocols., (© 2024 Wiley Periodicals LLC.)

Aim: To evaluate the effect of chlorhexidine gluconate-loaded phase-transited lysozyme (CHG@PTL) coating on inhibiting bacterial adhesion and biofilm formation in an ex vivo root canal dentine model., Methodology: The physicochemical and structural characteristics of CHG@PTL nanoparticle suspension and its coating formed on the dentine surface were analysed by thioflavin T fluorescence assay, transmission electron microscopy and confocal laser scanning microscopy (CLSM). The sustained chlorhexidine release profile of the CHG@PTL coating on the dentine surface was compared with that of the 2% CHG solution. By comparing with phosphate-buffered saline, 1% sodium hypochlorite and 2% CHG solutions, the sustained antibacterial ability of the CHG@PTL coating and its effects on adhesion and biofilm formation of three types of bacteria (E. faecalis, S. mutans, and A. viscous) were analysed in ex vivo root canal dentine models using the serial plate transfer test (SPTT) and CLSM with live/dead bacterial staining, respectively., Results: CHG promoted the lysozyme protein to form a higher proportion of β-sheet structure during phase transition. In the CHG@PTL nanoparticle suspension, characteristic drug-loaded nanospheres with a high concentration of CHG molecules inside and an outer PTL nanofilm were observed, and they formed a thinner and tighter coating on the dentine surface. The CHG@PTL coating on the dentine surface showed a significantly higher cumulative release amount of chlorhexidine than that of 2% CHG (p < .05). The results of SPTT showed that the CHG@PTL coating had a longer antibacterial duration than the control groups. After 12 h of incubation, a higher number of bacteria were agglutinated on the CHG@PTL coating surface compared to the control groups (p < .05). After 7 days of incubation, the number of agglutinated bacteria significantly decreased. At two time points, the percentage of dead bacteria on the CHG@PTL coating surface was the highest among all experimental groups based on CLSM observation (over 99.9% for all three bacteria, p < .001)., Conclusions: CHG@PTL nanoparticle suspension could form an antimicrobial coating on the surface of dentine with a novel 'agglutinating bacteria and sterilizing' mode., (© 2024 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Objectives: The aim of this in-vitro study was to investigate the masking efficacy of adhesively sealed enamel cracks with resin infiltration compared with the use of a universal adhesive., Methods: Enamel cracks were induced on the labial surfaces of bovine teeth using a pendulum impact tester. Specimens were treated adhesively by resin infiltration with ICON (DMG) or Scotchbond Universal Plus (3M). Standardized digital images were taken pre- and postoperatively by three methods: flashlight from the front, transmitted light from behind, and DiagnoCam (KaVo). Four calibrated raters performed the qualitative visual evaluation of all images of each type: severely visible enamel cracks (1), clearly visible enamel cracks (2), slightly visible/aesthetically acceptable enamel cracks (3) and no visible enamel cracks (4). Specimens were selected to measure infiltration depth by confocal microscopy., Results: Postoperatively, the masking efficacy in teeth with enamel cracks was significantly higher using ICON compared with Scotchbond Universal Plus in all groups (p < 0.001). Infiltration depths of ICON were significantly higher compared to those of Scotchbond Universal Plus (p < 0.002)., Conclusions: Resin infiltration offers a straightforward and effective treatment option for masking trauma-induced enamel cracks, demonstrating superior efficacy over adhesive sealing. Further studies are necessary to evaluate the long-term stability of the optical improvements achieved through resin infiltration., Clinical Significance: Resin infiltration might pose a therapeutic option for clinicians to enhance the aesthetic appearance of trauma-induced enamel cracks., Competing Interests: Declaration of competing interest H.M.-L. is appointed as an inventor for patents of an infiltration technique for dental caries lesions, held by Charité-Universitätsmedizin Berlin, and receives royalties from DMG, the manufacturer of Icon. All other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The formation of biofilms enhances bacterial antibiotic resistance, posing significant challenges to clinical treatment. Methicillin-resistant Staphylococcus aureus (MRSA) is a primary pathogen in biofilm-associated infections. Its high antibiotic resistance and incidence rates make it a major clinical challenge, underscoring the urgent need for novel therapeutic strategies. Building on previous research, this study employs nanotechnology to fabricate dihydroartemisinin-chitosan nanoparticles (DHA-CS NPs) and, for the first time, applies them to the treatment of MRSA biofilm infections. The antibacterial and anti-biofilm activities of these compounds were evaluated, and their potential mechanisms of action were preliminarily explored. The results demonstrated that the DHA-CS NPs exhibited a minimum inhibitory concentration (MIC) of15 μg/mLand a minimum bactericidal concentration (MBC) of 30 μg/mL. At 15 μg/mL, the DHA-CS NPs significantly inhibited MRSA biofilm formation (P < 0.001),while at 7.5 μg/mL, they dispersed 67.4 ± 3.77 % of the preformed biofilms (P < 0.001). Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) confirmed the disruption of MRSA biofilms. Mechanistic studies, including phenol-sulfuric acid assays, static biofilm microtiter plate assays, and RT-qPCR, revealed that the DHA-CS NPs inhibited the synthesis of extracellular polymeric substances (EPS), suppressed the release of extracellular DNA (eDNA), and downregulated key biofilm-related genes (icaA, sarA, cidA, and agrA). These findings suggest that DHA-CS NPs hold significant promise for inhibiting and eradicating MRSA biofilms, providing a theoretical basis for the development of novel antibiofilm therapies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The colonization of dental implants by oral biofilms causes inflammatory reactions that can ultimately lead to implant loss. Therefore, safety-integrated implant surfaces are under development that aim to detect bacterial attachment at an early stage and subsequently release antibacterial compounds to prevent their accumulation. Since primary oral colonizers ferment carbohydrates leading to local acidification, pH is considered a promising trigger for these surfaces. As a prerequisite for such systems, the present study aimed at specifically analyzing the pH at the interface between implant material and oral biofilms. For this purpose, in vitro-grown Streptococcus oralis monospecies biofilms and an established multispecies biofilm on titanium discs as well as in situ-grown biofilms from orally exposed titanium-equipped splints were used. Mature biofilm morphology was characterized by live/dead fluorescence staining, revealing improved growth from in vitro to in situ biofilms as well as a general decreasing membrane permeability over time due to the static incubation conditions. For pH analysis, the pH-sensitive dye C-SNARF-4 combined with 3-dimensional imaging by confocal laser-scanning microscopy and digital image analysis were used to detect extracellular pH values in different biofilm layers. All mature biofilms showed a pH gradient, with the lowest values at the material interface. Interestingly, the exact values depicted a time- and nutrient-dependent gradual acidification independently of the biofilm source and for in situ biofilms also independently of the sample donor. After short incubation times, a mild acidification to approximately pH 6.3 could be observed. But when sufficient nutrients were processed for a longer period of time, acidification intensified, leading to approximately pH 5.0. This not only defines the required turning point of pH-triggered implant release systems but also reveals the opportunity for a tailored release at different stages of biofilm formation., Competing Interests: Declaration of Conflicting InterestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Objective: This study investigated the associations among endodontic instruments, ultrasonic tips and various final irrigation protocols for removing intracanal and intratubular biofilms in long oval canals., Methodology: One hundred mandibular premolars inoculated with Enterococcus faecalis were divided into two groups: the control group (CG: n = 10), which received no treatment; and the test groups (n = 30), which included saline (SS), sodium hypochlorite (2.5% NaOCl) and chlorhexidine (2% CHX). The samples were prepared with Reciproc ® Blue 25/0.08, Flatsonic, Clearsonic and Reciproc Blue 40/0.06. Each test group was divided into three subgroups (n = 10): conventional irrigation (CI) with syringes and needles, passive ultrasonic irrigation (PUI) and continuous ultrasonic irrigation (CUI) with an Irrisonic tip. The root canals were examined using confocal laser scanning microscopy. Mann‒Whitney and Kruskal‒Wallis tests followed by Dunn post hoc tests (p < 0.05) were used for statistical analysis., Results: NaOCl and CHX were effective for intracanal and intratubular decontamination. Saline showed significant decontamination in the intratubular areas when CUI was used. The CI method was the least effective for intracanal decontamination., Conclusion: The use of Flatsonic and Clearsonic ultrasonic tips is promising for root canal disinfection., Clinical Relevance: The disinfection techniques in endodontic treatment focus on the removal of biofilms from long oval canals, which is crucial for effective cleaning during root canal procedures., Competing Interests: Declarations. Ethical approval: All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Human teeth were used after Ethics Committee Approval (number: 50179321.9.0000.5417). Informed consent: Informed consent was obtained from all individual participants included in the study. Competing interests: The authors declare no competing interests., (© 2025. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Measurement of glycated hemoglobin (HbA1c) in human red blood cells plays a critical role in the diagnosis and treatment of diabetes mellitus. However, recent studies have suggested large variation in the relationship between average glucose levels and HbA1c, creating the need to understand glucose variability at the cellular level. Here, we devised a fluorescence-based method to quantitatively observe GLUT1-mediated intracellular glucose analog tracer uptake in individual RBCs utilizing microfluidics and confocal microscopy. For the first time, we demonstrate that intracellular/extracellular tracer percentages can be measured at the single cell level using the fluorescently labeled glucose analog, 2-NBDG. A small donor panel study indicates that the characteristic intracellular 2-NBDG percentages can statistically differ based on race (i.e., Caucasian/Hispanic vs Black). RBC intracellular glucose analog tracer 2-NBDG levels show significant variability both from cell-to-cell and from donor-to-donor. This specific transport mechanism will affect HbA1c formation in erythrocytes. This finding further supports a more personalized, and perhaps improved, diagnostic strategy for diabetes., Competing Interests: Competing interests: All authors are current or former employees of Abbott Laboratories., (© 2024. The Author(s).)

Introduction: To evaluate the bacterial biofilm, smear layer and debris removal efficacy of a hydro-dynamic cavitation system with physiological saline using a new ex vivo model., Methods: Seventy-five dentin discs were prepared from fifty-four extracted teeth. Seventy-five artificial root sockets were prepared. Sixty dentin discs were used to grow 3-week-old bacterial biofilms, while smear layer and debris were produced on fifteen dentin discs. These dentin discs were adhered to the middle third of the artificial root canals. The sixty ex vivo models with biofilm-covered dentin discs were divided into six groups: control, needle with physiological saline, ultrasonic with physiological saline, Odne™ Clean with physiological saline, needle with 3% NaOCl, and ultrasonic with 3% NaOCl. Biofilm removal efficacy was evaluated using confocal laser scanning microscopy. The fifteen ex vivo models with smear layer and debris-covered dentin discs were divided into three groups: control, Odne™ Clean with physiological saline, and 5% NaOCl followed by 17% EDTA. Smear layer and debris removal efficacy was evaluated using scanning electron microscopy. Statistical analysis was performed using one-way analysis of variance for comparisons involving more than two groups. Post-hoc pairwise comparisons were conducted using the Tukey test., Results: Odne TM Clean with physiological saline (98%) performed significantly better than needle irrigation (47%) or ultrasonic activation (54%) with physiological saline (P < 0.05). Odne TM Clean with physiological saline removed biofilms as effectively as needle irrigation (97%) or ultrasonic activation (98%) with 3% NaOCl (P > 0.05). Additionally, 5% NaOCl followed by 17% EDTA (score: 1.33) removed the smear layer significantly better than Odne TM Clean with physiological saline (score: 4.47) (P < 0.05). However, Odne TM Clean with physiological saline (score: 1.27) removed debris as effectively as 5% NaOCl followed by 17% EDTA (score: 1.13) (P > 0.05)., Conclusions: Odne TM Clean with physiological saline can effectively remove bacterial biofilm and debris from the dentin surface but cannot effectively remove the smear layer. Utilizing Odne TM Clean during the final irrigation may enhance root canal cleaning efficacy., Competing Interests: Declarations. Ethics approval and consent to participate: The clinical Research Ethics committee of the University of British Columbia (certificate H1202430) approved the protocol of this study. All participants provided signed informed consent, authorizing the use of their extracted premolars for research purposes. The authors ensure that all procedures were conducted in line with relevant guidelines and regulations. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: Cellular vacuolization is a commonly observed phenomenon under physiological and pathological conditions. However, the mechanisms underlying vacuole formation remain largely unresolved., Methods: LysoTracker Deep Red probes and Enhanced Green Fluorescent Protein-tagged light chain 3B (LC3B) plasmids were employed to differentiate the types of massive vacuoles. By confocal microscopy, lysosome-like massive vacuoles (LysoTracker Deep Red + ), autophagosome-like massive vacuoles (LC3B-enhanced green fluorescent protein (EGFP + )), and autolysosome-like massive vacuoles (LC3B-EGFP + LysoTracker Deep Red + ) in starved HEK293T cells were observed., Results: In this study, we demonstrated that nutrient deficiency can induce the formation of massive vacuoles that appear highly electron-lucent in HEK293T cells. Additionally, these massive vacuoles, resulting from nutrient depletion, can originate from various organelles, including small vacuoles, autophagosomes, lysosomes, and autolysosomes. We found that massive vacuoles could form through two primary mechanisms: the accumulation of small vacuoles into larger vacuoles or the fusion of homogeneous or heterogeneous vacuoles. Further analysis revealed that the membranes of massive vacuoles, regardless of origin, were composed of a bilayer membrane structure. As the volume of the massive vacuoles increased, the cytoplasm and nucleus were displaced toward the periphery of the cells, leading to the formation of signet ring-like cells. Interestingly, we provided evidence that complete massive vacuoles or autophagosome-like massive vacuoles can be released and exist independently outside HEK293T cells., Conclusions: Nutrient deprivation induces the formation of heterogeneous, massive vacuoles in human embryonic kidney cells, some of which contribute to the development of signet ring cells, while others lead to extracellular vacuole formation., (© 2025 The Author(s). Published by IMR Press.)

This study set out to characterise the in vitro development, including musculature, of the microphallid parasite of the barnacle Semibalanus balanoides (Linnaeus, 1767), Maritrema gratiosum Nicoll, 1907 collected in Scotland. An in vitro culture model was developed to obtain ovigerous adults of M. gratiosum and their morphology was observed. Different media were tested and NCTC-109 was chosen as the best medium. The effects of different concentrations of serum upon adult longevity, size and egg production was measured. Survival for 10-days was achieved when flukes were cultured in NCTC-109 plus chicken serum and antibiotics. Forty percent chicken serum seemed to provide better results in terms of survival time and producing flukes with the largest body lengths. Both normal and abnormal eggs were observed from adults cultured in vitro. Confocal microscopy was undertaken to provide details of the development of the parasite's ultrastructure, including musculature, during the course of in vitro culture. While the musculature of M. gratiosum was similar to that of other microphallids, some additional novel structures were observed, most notably a ligament connecting pars prostatica and seminal vesicle and a racket-shaped excretory bladder. This study has provided greater insight into the biology M. gratiosum, and also developed a good in vitro model which might be applied to ecological or medical research in the future., Competing Interests: Declarations. Ethical approval: No ethical approval was required for the work with barnacles; however, our practices align with ethical principles regarding the collection and treatment of animals and adhere to the animal welfare regulations in the University of Stirling and the UK. Consent to participate: All authors consent to participate in this publication. Consent for publication: All authors consent to publish the manuscript. Competing interests: The authors declare no competing interests. Research data: Available on request by contacting the lead author. Clinical trial: Clinical trial number: not applicable., (© 2025. The Author(s).)

Thiols function as antioxidants in food, prolonging shelf life and enhancing flavor. Moreover, thiols are vital biomolecules involved in enzyme activity, cellular signal transduction, and protein folding among critical biological processes. In this paper, the fluorescent probe PYL-NBD was designed and synthesized, which utilized the fluorescent molecule pyrazoline, the lysosome-targeted morpholine moiety, and the sensing moiety NBD. Probe PYL-NBD was tailored for the recognition of biothiols through single-wavelength excitation, yielding distinct fluorescence emission signals: blue for Cys, Hcy, and GSH; green for Cys, Hcy. Probe PYL-NBD exhibited rapid reaction kinetics (<10 min), distinct fluorescence response signals, and low detection limits (15.7 nM for Cys, 14.4 nM for Hcy, and 12.6 nM for GSH). Probe PYL-NBD enabled quantitative determination of Cys content in food samples and L-cysteine capsules. Furthermore, probe PYL-NBD had been successfully applied for confocal imaging with dual-channel detection of biothiols in various biological specimens, including HeLa cells, zebrafish, tumor sections, and Arabidopsis thaliana., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Streptococcus mutans (S. mutans) contributes to caries. The biofilm formed by S. mutans exhibits greater resistance to drugs and host immune defenses than the planktonic form of the bacteria. The objective of this study was to evaluate the anti-biofilm effect of cordycepin from the perspective of metabolomics., Methods: The minimum inhibitory concentration (MIC) was determined to evaluate the antimicrobial effect of cordycepin on planktonic S. mutans. The 24-h biofilm was treated with 128 µg/mL of cordycepin for 10 min at the 8- or 20-h time points. Biofilm biomass and metabolism were assessed using crystal violet and MTT assays and cordycepin cytotoxicity was evaluated in human oral keratinocytes (HOK) using CCK-8 assays. The live bacterial rate and the biofilm volume were assessed by confocal laser scanning microscopy. Metabolic changes in the biofilm collected at different times during with cordycepin were analyzed by metabolomics and verified by quantitative real-time PCR., Results: The results showed that treatment with 128 µg/mL cordycepin reduced both the biomass and metabolic activity of the biofilm without killing the bacteria, and cordycepin at this concentration showed good biocompatibility. Metabolomics analysis showed that differentially abundant metabolites following cordycepin treatment were mainly related to purine and nucleotide metabolism. After immediate treatment with cordycepin, genes related to purine and nucleotide metabolism were downregulated, and the levels of various metabolites changed significantly. However, the effect was reversible. After continuing culture for 4 h, the changes in genes and most metabolites were reversed, although the levels of 2'-deoxyadenosine, 2'-deoxyinosine, and adenine remained significantly different., Conclusions: Cordycepin has the effect of anti-biofilm of S. mutans, mainly related to purine and nucleotide metabolism., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Purpose: This study investigated the synergistic effects of reduced graphene oxide (RGO) on the antibacterial activity of three calcium hydroxide-based intracanal medicaments with different vehicles., Methods: Multispecies biofilms were cultured in a bovine root canal model. Intracanal medicaments containing nonaqueous vehicles, including N-methyl-2-pyrrolidone (NMP; CleaniCal), propylene glycol (PG; UltraCal XS), and polyethylene glycol (PEG; Calcipex II), were placed in the model. The synergistic effects of RGO were evaluated by analyzing colony-forming units, extracellular deoxyribonucleic acid (eDNA) levels, and findings from confocal laser-scanning microscopy (CLSM) and scanning electron microscopy. To evaluate the cytotoxicity of each vehicle, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed. To determine whether RGO altered the physicochemical properties of the medicaments, flowability and pH were measured. A P-value of <0.05 was considered to indicate statistical significance., Results: In the CleaniCal group, bacterial viability, eDNA levels, and extracellular polymeric substrate (EPS) were significantly lower in the presence of RGO. However, only bacterial viability significantly differed in the other groups. PEG resulted in the highest cell viability among the vehicles. Furthermore, there was no significant difference in flowability or pH in relation to the inclusion of RGO., Conclusion: The results suggest that RGO enhances the antibacterial effect of intracanal medicaments containing NMP.

The excessive production and use of plastics increase the release of micro- and nanoplastics (MNPs) into the environment. In recent years, research has focused on the occurrence of MNPs in air, soil and water. Nevertheless, there is still a lack of knowledge regarding MNPs in plants. To determine the load, translocation of MNPs and their effects on metabolism, pak choi, tomato, radish and asparagus have been exposed with fluorescent-labelled poly(methyl methacrylate) or polystyrene (PS) MNPs. The entry of nanoparticles (NPs) of various sizes (100-500 nm) and surface modifications (unmodified, COOH or NH 2 ) into plants has been demonstrated using confocal laser scanning microscopy (CLSM). The translocalization from root to shoot and the accumulation of NP in the intercellular spaces were regardless of the surface modification. In addition, metabolomics was used to evaluate metabolic changes induced by MNPs in pak choi. Changes in phenolic compounds, phytohormone derivatives and other classes of compounds known to be triggered by various environmental stresses have been identified. The present study demonstrates the uptake and translocalization of MNPs in edible parts of vegetables and may pose a hazard for humans., (© 2024 The Author(s). Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Objectives: The absorption of biostimulatory particulate matter following its application to fractional skin defects remains poorly understood, and even less is known about its in vivo impact in terms of tissue integration. The objectives of this study are twofold: (1) to evaluate the potential of calcium hydroxylapatite (CaHA) to penetrate through skin treated with a fractional laser; and (2) to assess the effectiveness of clinical laser scanning microscopy technologies in monitoring the effects of such treatment over time., Methods: One area on a volunteer's arm was treated with a fractional erbium laser (Sciton Inc., Palo Alto, CA), while a second area received the same laser treatment followed by CaHA topical application. We used reflectance confocal microscopy (RCM) and multiphoton microscopy (MPM) to noninvasively image beneath the surface of the treated skin to study and monitor the effects of these treatments within 1 h of treatment and at four additional time points over a 6-week period., Results: One hour posttreatment, at different depths beneath the skin surface, MPM and RCM provided similar visualizations of laser-induced channels. In skin treated by both laser and CaHA, these two imaging methods provided complementary information. RCM captured the lateral and depth distribution of CaHA microspheres and were seen as bright spheres as they became incorporated into the healing tissue. MPM, meanwhile, visualized the CaHA microparticles as dark shadow spheres within the laser-induced channels and encroaching healing tissue. Furthermore, MPM provided critical information about collagen regeneration around the microspheres, with the collagen visually marked by its distinct second harmonic generation (SHG) signal., Conclusions: This observational pilot study demonstrates that CaHA, a collagen stimulator used as a dermal filler, can not only be inserted into the dermis after fractional laser treatment but remains in the healing skin for at least 6 weeks posttreatment. The noninvasive imaging techniques RCM and MPM successfully captured the presence of CaHA microspheres mid-dermis during the healing phase. They also demonstrated new collagen production around the microspheres, highlighting the effectiveness of these imaging approaches in monitoring such treatment over time., (© 2024 Wiley Periodicals LLC.)

Objectives: Evaluate the remineralisation ability of experimental resin-based materials containing Fluoride-Doped Calcium Phosphate (FDCP), applied in mineral-deficient dentine in combination with a biomimetic dual-analogue primer., Methods: Artificial dentine carious lesions were created in occlusal cavities of human molars. An experimental resin-based adhesive and flowable composite, containing FDCP (10wt% and 20wt%, respectively), were applied (±) with a biomimetic primer containing polyacrylic acid and sodium tripolyphosphate. A glass-ionomer cement and a conventional adhesive/composite were used as comparative materials. All specimens were submitted to chewing simulation in artificial saliva (AS). Subsequently, these were cut into slabs and submitted to Knoop microhardness testing and Raman spectroscopy (AS: 0, 15, 30 and 60 days). Further specimens were analysed through confocal scanning microscopy (AS: 0, 60 days). Total demineralised dentine and disc-shape materials were analysed through FTIR, SEM and EDX (Ca/P ratio) after prolonged contact in AS. Data were statistically analysed (two-way ANOVA and multiple comparison Bonferroni; α=0.05)., Results: Only the experimental FDCP materials induced a significant increase in microhardness after storage in AS, especially in those specimens pre-treated with the biomimetic primer, except at 50 µm underneath the interface. Raman and CLSM analysis corroborated such results. The ability of the experimental FDCP resin-based materials to induce apatite-like deposition was demonstrated through SEM-EDX and FTIR., Conclusions: Resin-based materials containing FDCP can mineralise the mineral-deficient dentine, especially when combined with a biomimetic dual-analogue primer., Clinical Significance: Such Innovative ion-releasing restorative systems applied after selective caries removal might promote remineralisation of fully and partially demineralised dentine and increase the durability of dental restorations., Competing Interests: Declaration of competing interest We wish to confirm that there are no known conflicts of interest associated with this publication, and there has been no significant financial support for this work that could have influenced its outcome., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Objective: To develop a specific methodology for identifying dental chipping and determining its temporal occurrence in past populations., Design: The analysed sample comprised of 2191 human teeth from various Bronze Age on the Iberian Peninsula (Argar culture, 1900-1450 cal BC). Among these, 471 chipped teeth were identified. Chipping was examined using various microscopic techniques (digital three-dimensional, optical, and confocal), focusing on distribution, morphology, position in the tooth, extent of damage, and post-chipping antemortem modifications (PCAM)., Results: The distribution and morphology of the chips enabled the identification chipping mechanism of the chipping, providing valid criteria to distinguish between antemortem and postmortem chipping. Microscopic analyses of the chipping segments-edges, sidewalls, surface, and surrounding area-facilitated determination of the time the chip ocurred (antemortem: recent, less recent, or not recent)., Conclusions: While experimental studies provide valuable insights into chipping mechanisms, many criteria may not be applicable to past populations because of the presence of PCAM. The lack of PCAM in some Argaric teeth suggests that previous studies may have underestimated the prevalence of chipping in past populations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

DFAT cells represent an attractive source of stem cells in tissue engineering and in the potential treatment of several clinical conditions. Our objective was to determine whether DFAT cells originate from mature adipocytes and address whether contamination from the stromal vascular fraction (SVF) could be as a source for these cells. A murine adiponectin-creERT;mT/mG model was used with the excision of the cassette induced by tamoxifen injection for the cells expressing adiponectin (adipoq). This model allows distinguishing of mature adipocytes (green fluorescence) from other SVF cell types (red fluorescence) based on the fluorescent protein expressed. Mature adipocytes and SVF cells were isolated from adipose tissues by collagenase digestion. Ceiling cultures were imaged by time-lapse microscopy. Confocal microscopy was used to follow cells over 21 days. Time-lapse microscopy experiments showed liposecretion occurring in mature adipocytes displaying green fluorescence. Confocal imaging allowed the identification of a heterogeneous cell population expressing green but also red fluorescence after 21 days of culture. Asymmetrical division of mature adipocytes was not observed. In conclusion, liposecretion of mature adipocytes is a phenomenon that can be observed in vitro and DFAT cells do originate from mature adipocytes . However, the population of DFAT cells is heterogenous., Competing Interests: AT receives research funding from Johnson & Johnson, Medtronic, GI Windows and Biotwin for studies on obesity or bariatric surgery and acted as a consultant for Bausch Health, Novo Nordisk, and Biotwin.

Purpose: To describe features in silicone oil keratopathy using multimodal imaging and histopathological examination., Methods: Case report., Result: A 21-year-old male developed right corneal decompensation in the heavy SO (HSO)-filled eye. The patient underwent an initial lensectomy, pars plana vitrectomy (PPV) and HSO tamponade due open-globe injury with corneal wound, lens damage and in two retained intravitreal glass foreign bodies, followed by a revisional PPV with HSO tamponade due to tractional detachment associated with proliferative vitreoretinopathy and epiretinal membrane. One month after the removal of HSO, ophthalmic examination of the right eye showed corneal decompensation. The AS-OCT showed corneal thickening, intrastromal scattered hyperreflective dots and large rounded/oval hyporeflective space; the latter were suggestive of emulsified HSO microbubbles and larger bubbles, respectively. In vivo confocal microscopy showed multiple presumed SO-related corneal changes, including hyper-reflective fibrotic changes in the basal epithelium, reduced density ans altered morphology of keratocytes cell population, increased pleomorphism and polymegathism of the endothelium with reduced endothelial cell, and presence of inflammatory cells. The patient underwent a penetrating keratoplasty, pupilloplasty and retropupillary iris-claw IOL implantation. The histopathological examination of the host corneal button showed Descemet's membrane irregularity and thickened corneal stroma with focal intrastromal silicone oil vacuoles, surrounded by macrophages., Conclusion: We described for the first time intrastromal hyperreflective dots as a sign associated with SO-related keratopathy. Moreover, this case report supports the ability of emulsified SO to penetrate the cornea inducing a local low-grade chronic inflammation., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Recent gene expression studies have revealed about 10 different states of microglia, some of which are characteristic for Alzheimer-like amyloid plaque pathology. However, it is not presently known how these translate into morphological features that would reflect microglia interaction with amyloid plaques. With optimized conditions for confocal microscopy in amyloid plaque forming APP/PS1 transgenic mice we reveal new details of how microglia processes interact with amyloid plaques. The microglia contacts differed drastically between purely diffuse plaque and those with a fibrillar core. We identified microglia that extend their enlarged processes through the diffuse shell of the amyloid plaques and cover the fibrillar plaque core with snowplow-like expanded end-feet. These end-feet were filled with the lysosomal marker CD68, while both non-fibrillar and fibrillar amyloid was found in perinuclear vesicles of some "snowplower" microglia. In the organized dense-core plaques, we consistently saw a layer of Apolipoprotein E (ApoE) between the fibrillar core and the microglial end-feet. ApoE covered also loose fibrillar amyloid and diffuse amyloid plaques that were about 10 μm or larger in diameter. These findings are compatible with both amyloid plaque phagocytosis and compaction by microglia. Further, they support a chemotactic role of ApoE for microglia contacts with amyloid plaques., (© 2024 The Author(s). GLIA published by Wiley Periodicals LLC.)

Background: The advent of ablative fractional photothermolysis has revolutionized laser dermatology by providing a method to produce well-standardized, precise, and repeatable microscopic lesions. These wounds typically heal within 1-3 weeks, depending on the body site, with a minimal risk of permanent scarring. This positions ablative fractional photothermolysis as an exemplary in vivo model for studying the skin's wound healing processes., Objectives: This study aims to evaluate and compare the effectiveness of two noninvasive imaging techniques, reflectance confocal microscopy (RCM) and line-field confocal optical coherence tomography (LC-OCT), in assessing skin wound healing following microscopic injuries induced by ablative fractional photothermolysis., Methods: The forearms of participating volunteers were treated and ablated with a CO 2 -Laser in a fractional pattern using varying power settings (2.5-10 mJ/MTZ). In vivo RCM and LC-OCT images were obtained at predefined time intervals post-laser treatment, ranging from 6 h to 14 days., Results: Vertical visualization of the lesions through both imaging modalities revealed a healing process characterized by the upward and outward movement of microscopic epidermal necrotic debris, thereby reducing the depth of the injury while forming an external crust. LC-OCT imaging demonstrated more comprehensive results with fewer movement artifacts. Conversely, horizontal visualization with both techniques highlighted a gathering of keratinocytes around the wounds, indicating the initiation of the regenerative process. RCM provided superior image clarity in this horizontal plane., Conclusions: RCM and LC-OCT offer valuable and complementary noninvasive alternatives to conventional biopsy methods for the assessment and characterization of the skin's wound healing process post-ablative fractional photothermolysis. These findings underscore the potential of such imaging techniques in enhancing our understanding of the wound healing process., Trial Registration: ClinicalTrials.gov identifier: NCT05614557., (© 2024 The Author(s). Lasers in Surgery and Medicine published by Wiley Periodicals LLC.)

Purpose: Corneal neurotization (CN) is a novel, potentially curative surgical procedure for the treatment of neurothophic keratopathy (NK). Patients with severe NK can present with corneal opacification requiring optical keratoplasty, which would likely fail without a proper trophic support of corneal nerves in the recipient cornea., Methods: This is a pilot study on 4 patients undergoing keratoplasty after CN. Pre- and postoperative data at 12, 24 months and at the last follow-up were collected for the examination of (i) best corrected visual acuity (BCVA), (ii) slit lamp examination and photograph acquisition with and without fluorescein staining, (iii) corneal aesthesiometry, (iv) in vivo confocal microscopy of the central cornea. Neurophysiological study of the corneal reflex before corneal graft and at last follow up was performed., Results: Four female patients (47.25 ± 5.06 y.o.) underwent keratoplasty after CN (3 penetrating keratoplasty, 1 deep anterior lamellar keratoplasty). The mean interval between CN and keratoplasty was 22 (± 12) months. The mean graft survival time was 42 (± 25) months. Graft follow-up ranged from 72 to 132 months. At the final follow-up, BCVA was improved in 2 out of 4 patients. The mean corneal sensitivity was 11.9 ± 8.3 mm at last follow-up. In vivo confocal microscopy confirmed the presence of functioning nerves at the last follow-up in all patients. NK-related complications occurred in 3 eyes (2 persistent epithelial defect, 1 corneal melting). The former complication was successfully treated by autologous serum eye drops while the latter required repeated keratoplasty., Conclusions: Keratoplasty is a viable strategy to improve visual acuity in patients with corneal opacity who underwent CN for the treatment of NK. Even in the presence of functioning corneal nerves before keratoplasty, surgeons should be aware of the increased rate of NK-related complications that could require the need for repeated procedure.

Different concentrations and infiltration times of rose bengal (Rb) were assessed for their impact on penetration depth and crosslinking efficacy in rabbit sclera. Fresh rabbit eyes were used. Rb solution with concentrations of 0.1%-0.9% were applied for 5-30 min to infiltrate the sclera. The penetration depth of Rb was observed with confocal microscopy. After infiltration, the sclera was irradiated by green light for crosslinking. The sclera's biomechanical stiffness and the resistance to enzyme digestion post-treatment were evaluated. Histopathological analysis and transmission electron microscopy were performed to observe the morphology. As the infiltration time increased, the penetration depth and the fluorescence intensity of the Rb in sclera increased. After 32 h, 48.6% of the scleral tissue was undissolved in the 0.5% Rb-10min group, followed by the 0.1% Rb-20min group (13.8%) and 0.05% Rb-30min group (7.7%). At 8% strain, the Young's modulus of the 0.05%Rb-30min, the 0.1% Rb-20min and the 0.5% Rb-10min group were respectively 1.77, 2.45 and 3.19 times greater than that of the untreated group. There were no morphological differences between the experimental group and the untreated group. RG-SXL significantly increased the diameter of large collagen fibers in the middle and inner layers of the sclera. Ultimately, 0.5% Rb infiltration for 10 min achieves an appropriate infiltration depth and crosslinking effect, and may thus be a feasible schedule for scleral crosslinking., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objectives: Caries is a significant public health challenge. Herein, novel tooth-targeting antimicrobial peptides (HABPs@AMPs) were developed by combining the antimicrobial peptide DJK-5 with hydroxyapatite (HA) binding peptides, providing a potential new strategy for caries management., Materials and Methods: The minimal inhibitory concentration (MIC 100 ) and minimal biofilm inhibitory concentration (MBIC 100 ) values of HABPs@AMPs were determined via micro-broth dilution and crystal violet staining. The affinities of the peptides for HA were measured by mass depletion, and the abilities of peptides to inhibit Streptococcus mutans (S. mutans) biofilm formation and kill 3-day-old S. mutans biofilms were evaluated in HA disk and tooth slice biofilm models through confocal laser scanning microscopy. Biocompatibility with human gingival fibroblasts was evaluated via CCK8 assays., Results: The best performing peptides, DJK-5@SVA and SVA@DJK-5 exhibited MIC 100 and MBIC 100 values of 31.25 µg/mL, similar to DJK-5. DJK-5@linker2@YSL had the highest affinity for HA, followed by YSL@DJK-5, DJK-5@linker1@YSL, and DJK-5@SVA. Moreover, the biofilms on HABPs@DJK-5 coated surfaces had more dead bacteria by volume than those in the DJK-5 and SVA groups (p < 0.05). DJK-5@SVA outperformed SVA@DJK-5 and DJK-5 in killing 3-day-old S. mutans biofilms (p < 0.05). With the exception of established biofilms on tooth slices, DJK-5@SVA exhibited greater killing efficiency in the bottom half of the biofilms than in the top half. The CCK-8 assay results confirmed peptides' biocompatibility., Conclusions: DJK-5@SVA with good affinity for HA, has excellent biocompatibility and efficacy against S. mutans biofilms., Clinical Relevance: HABPs@AMPs with effective inhibitory effects on the growth of S. mutans and biofilm formation, contributing to intraoral targeted application AMPs and providing a new strategy for caries management., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: This study aims to comparatively assess the preventive and protective effects of the self-assembling peptide P 11 -4 on enamel erosion and evaluate the potential for enamel surface recovery when professional products are combined with home-use dental-care products during the erosive process., Materials and Methods: Ninety-nine bovine incisors were divided into nine groups: a control group, four groups with the application of professional-products [P 11 -4 peptide (Curodont-Repair), stannous/Sn 2+ containing solution (8% Sn 2+ ), casein-phosphopeptide-amorphous-calcium-phosphate fluoride/CPP-ACPF (MI Varnish), sodium fluoride/NaF (Profluorid)] and four groups with the combination of professional products and home-use daily dental care products [P 11 -4 peptide (Curodont Repair + Curodont Protect), stannous ions containing agents (8% Sn 2+ +Emofluor Gel Intensive-Care), CPP-ACPF (MI Varnish + MI Paste Plus), NaF (Profluorid + ReminPro)]. Professional products were applied once before a five-day erosive cycle, involving six 2-minute citric-acid exposures per day. In combined-groups, a home-use dental-care product was applied daily at the end of each cycle day. After the cycle, lesion depth and fluorescence were analyzed using confocal-laser-scanning-microscopy, and results were statistically evaluated using paired t-tests, ANOVA, and Tukey's post-hoc tests., Results: The P 11 -4 group was significantly more effective among the professional-only groups in both parameters (p<0.001). All combined-groups were determined to be statistically more successful than their respective professional-groups in both parameters (p<0.05). Based on lesion depth data, it was observed that the P 11 -4 and stannous-containing combined-groups showed statistically similar results (p>0.05). In terms of lesion fluorescence data, the P 11 -4 combined-group was found to be statistically more successful than all other study groups (p<0.05)., Conclusion: The self-assembling peptide P 11 -4 was determined to exhibit the best preventive and protective anti-erosive effect in both professional and combined applications., Clinical Relevance: A positive relationship was observed between the support of professional applications with home-use daily-dental-care products and protection against erosive lesions., Competing Interests: Declarations. Ethical approval: This article does not contain any studies with human participants or animals performed by any authors. Informed consent: For this type of study, formal consent is not required. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The aim of this study was to assess roughness profile and surface roughness after simulated toothbrushing cycles, as well as the degree of conversion (DC) of bulk-fill resin composites at different depths. Forty nine composite discs were made from three low-viscosity bulk-fill resins (Filtek Bulk-Fill Flowable/3M Oral Care - FBF, Beautifil-Bulk Flowable/Shofu Inc. - BBF and Surefill SDR Flow/Dentsply Caulk - SDR), three high-viscosity bulk-fill resin composites (Filtek Bulk-Fill Restorative/3M Oral Care - FBR, Beautifil-Bulk Restorative/Shofu Inc. - BBR and Tetric EvoCeram Bulk Fill/Ivoclar Vivadent - TEB) and one conventional composite (Filtek Supreme Ultra/3M Oral Care - FSU) (n=7), using addition silicon molds (2 mm thick x 10 mm diameter). The roughness profile (Rv) and surface roughness (Sa) were analyzed after 30,000 brushing cycles using confocal laser scanning microscopy. Five samples (4 mm thick x 2 mm diameter) were produced per resin composite and light-cured for 20 seconds using a LED-curing unit. After 24 hours at 37oC, DC of samples was evaluated at four depths, using a confocal Raman microscope. The data were analyzed by ANOVA and Tukey post hoc multiple-comparison tests (α = 0.05). The Rv values of FSU were lower than those obtained for BBR and BBF. The toothbrushing significantly decreased the Sa for all composites. The DC was significantly reduced at 4 mm depth. Toothbrushing changed composite surface of all materials, especially for BBR and BBF. Composites were not able to maintain the DC values shown at the top compared with those at the bottom (4 mm depth).

Background: The purpose of this study was to compare AH Plus Bioceramic Sealer (AHPB), AH Plus (AHP), MTA Fillapex (MTAF), and Sure-Seal (SS) sealers for dentinal tubular penetration ability., Methods: Forty extracted human mandibular premolars were instrumented and divided into 4 groups based on the sealer used for filling (n = 10): AHPB, AHP, MTAF, SS. After filling the root canals, the roots were entombed in acrylic resin blocks and cut vertically in the long axis to obtain 1-mm-thick disks from 3 mm (at apical third), 8 mm (at middle third) and 13 mm (at coronal third) away the apical foramen. The images of 120 samples (30 for each group) were scanned by using confocal laser scanning microscopy and analyzed with the ImageJ software. Total penetration area and the root canal area were measured, and the ratio of these values were recorded as sealer penetration ratio. The data were evaluated for each third and in total using one-way ANOVA with post-hoc Tukey's test. The significance level was set to P < 0.05., Results: In the apical, middle and coronal thirds and in total, MTAF and AHP, and SS and AHPB had statistically significant identical penetration ratio values. Both SS and AHPB demonstrated statistically considerably greater penetration % as compared to MTAF and AHP in all thirds (P < 0.05), and in total (P < 0.05)., Conclusion: AHPB and SS provided a higher penetration % than AHP and MTAF in apical, middle and coronal thirds, and in total., Competing Interests: Declarations. Ethics approval and consent to participate: Ethical approval was received for this in vitro study from the Ankara Yıldırım Beyazıt University, Ethical Committee of Health Sciences. (Date: 13.04.2023, number: 2023/141). An informed consent was obtained from the owner(s) of the teeth to use the teeth. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests. Authorship decleration: All authors have contributed significantly, and all authors agree with the manuscript. Disclosure: Authors declare any financial support or relationships related to this study., (© 2024. The Author(s).)

Early cancer detection plays a vital role in improving the survival rate of cancer patients, underscoring the importance of developing cancer detection methods. However, it is a great challenge to achieve simple, rapid, and accurate methods for simultaneously discerning various cancers. Herein we developed a 5-element porphyrin-embedded dendrimer-based sensor array, targeting the parallel discrimination of multiple cancers. The porphyrin-embedded dendrimers were modified with various functional groups to generate differentiated interactions with diverse cancer cells, which has been validated by fluorescence responses and laser confocal microscopy imaging. The dual-channel, five-element array, featuring ten signal outputs, achieved 100% accuracy in distinguishing between one human normal cell and six human cancerous cells, as well as in differentiating among mixed cells. Moreover, the screen 6-channel array can accurately distinguish 9 cells from mice and humans in minutes through optimization by multiple machine learning algorithms, including two normal cells and 7 cancerous cells with only 1000 cells, highlighting the significant potential of a porphyrin-embedded dendrimer-based parallel discriminating platform in early cancer diagnosis.

Multilayer paper-based cell culture, as an in vitro three-dimensional (3D) cell culture method, has been frequently used to research drug bioavailability, therapeutic efficacy, and dose-limiting toxicity in malignant tumors. This paper proposes a heterogenous multilayer paper stacking co-culture system to establish a model of natural killer (NK) cells moving through the endothelium layer and attacking tumor spheroids. This system consists of three layers: a bottom tumor-spheroid layer, a middle invasion layer, and a top endothelium layer. NK-92 cells were placed in the supernatant on top of the three layers. After two days of co-culture, the attack of tumor spheroids by NK cells was observed. We additionally examined the infiltration of NK-92 cells within the tumor spheroids at different Z -axis depths using a confocal microscope, and the results suggested that the system successfully realizes NK cells traveling cross the endothelium layer to form tumor-infiltrating NK cells (TINKs). The potential application of multilayer paper for co-culture models involving cancer cells and immune cells holds great promise for exploring the interaction dynamics of these two cell types.

Laminopathies represent a wide range of genetic disorders caused by mutations in gene-encoding proteins of the nuclear lamina. Altered nuclear mechanics have been associated with laminopathies, given the key role of nuclear lamins as mechanosensitive proteins involved in the mechanotransduction process. To shed light on the nuclear partners cooperating with altered lamins, we focused on Src tyrosine kinase, known to phosphorylate proteins of the nuclear lamina. Here, we demonstrated a tight relationship between lamin A/C and Src in skin fibroblasts from two laminopathic patients, assessed by advanced imaging-based microscopy techniques. With confocal laser scanning and Stimulated Emission Depletion (STED) microscopy, a statistically significant higher co-distribution between the two proteins was observed in patients' fibroblasts. Furthermore, the time-domain fluorescence lifetime imaging microscopy, combined with Förster resonance energy transfer detection, demonstrated a decreased lifetime value of Src (as donor fluorophore) in the presence of lamin A/C (as acceptor dye) in double-stained fibroblast nuclei in both healthy cells and patients' cells, thereby indicating a molecular interaction that resulted significantly higher in laminopathic cells. All these results demonstrate a molecular interaction between Src and lamin A/C in healthy fibroblasts and their aberrant interaction in laminopathic nuclei, thus creating the possibilities of new diagnostic and therapeutic approaches for patients.

Objective: Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is an autoimmune neuropathy characterized by progressive or relapsing-remitting weakness and sensory deficits. This study aims to evaluate the utility of corneal confocal microscopy (CCM) in diagnosing and monitoring CIDP., Methods: We analysed 100 CIDP patients and 31 healthy controls using CCM to measure corneal nerve fiber density (CNFD), length (CNFL), and branch density (CNBD). Standardized clinical and electroneurographic evaluation were conducted, and statistical analyses were performed to compare CCM parameters between groups and across disease stages., Results: CIDP patients and subgroups exhibited significant reduction in CNFD, CNFL, and CNBD compared to controls. This reduction was observed in late disease stages and severe overall disability sum score (ODSS), and Inflammatory Neuropathy Cause and Treatment Sensory Sum Score (ISS). CCM parameters correlated with axonal pathology in electroneurography of sensory, but not motor nerves. Despite the significant differences, the diagnostic sensitivity (41%) and specificity (77%) of CCM parameters were limited., Conclusion: While CCM effectively differentiates CIDP patients from healthy controls and was associated with disease severity, its diagnostic accuracy for routine clinical use is a posteriori. However, CCM shows promise as a non-invasive tool for monitoring sensory axonal pathology in CIDP., Competing Interests: Declarations. Conflict of interest: The authors declare that they have no conflict of interest. Ethical approval: All procedures performed in studies involving human participants were in accordance with the ethical standard of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendment or comparable ethical standards. The study was approved by the local ethics committee. Patient and public involvement: The patients gave spoken and written consent for publication and participation to the study. Disclosures: Rafael Klimas: received research funding from The LFB Group France and Ruhr-University, Bochum and travel funding from Grifols and Takeda; not related to this work. Dietrich Sturm: nothing to disclose. Annika Altenborg: nothing to disclose. Nayia Stylianou: nothing to disclose. Sophie Huckemann: nothing to disclose. Zornitsa Gasz: nothing to disclose. Jörg Philipps: nothing to disclose. Thomas Grüter: received travel funding and speaker honoraria from CSL Behring, Sanofi, Novartis, Biogen Idec, Bristol Myers Squibb, and Viatris, none related to this work. Greiner Tineke: nothing to disclose. Lynn Eitner: nothing to disclose. Tineke Greiner: nothing to disclose. Christoph Meier: nothing to disclose. Elena Enax-Krumova: nothing to disclose. Matthias Vorgerd: nothing to disclose. Peter Schwenkreis: received speaker honoraria from Merck Healthcare and Novartis Pharma, and funding from Berufsgenossenschaft für Gesundheitsdienst und Wohlfahrtspflege (BGW), none related to this work. Ralf Gold: serves on scientific advisory boards for Teva Pharmaceutical Industries Ltd., Biogen Idec, Bayer Schering Pharma, and Novartis; has received speaker honoraria from Biogen Idec, Teva Pharmaceutical Industries Ltd., Bayer Schering Pharma, and Novartis; serves as editor for Therapeutic Advances in Neurological Diseases and on the editorial boards of Experimental Neurology and the Journal of Neuroimmunology; and receives research support from Teva Pharmaceutical Industries Ltd., Biogen Idec, Bayer Schering Pharma, Genzyme, Merck Serono, and Novartis, none related to this manuscript. Anna Lena Fisse: received research funding by Georgius Agricola Stiftung Ruhr and Ruhr-University, Bochum (FoRUM-program), received honoraria and travel grants from Novartis AG, Sanofi and Eisai GmbH, none related to this work. Owns shares of Fresenius SE & Co., Gilead Sciences, Medtronic PLC and Novartis AG. Jeremias Motte: received travel grants from Biogen idec, Novartis AG, Teva and Eisai GmbH, his research is funded by Klaus Tschira Foundation and Ruhr-University, Bochum (FoRUM-program); none related to this work. Kalliopi Pitarokoili: received travel funding and speaker honoraria from Biogen Idec, Novartis and Bayer Schering Pharma and funding from the Ruhr-University, Bochum (FORUM-Program), none related to this work., (© 2024. The Author(s).)

Objectives: Biofilm is the major challenge in chronic wound management. Instilling a wound cleansing solution aids in wound bed cleaning and infectious pathogen elimination. Negative pressure wound therapy (NPWT) improves the wound-healing process. This study investigated the efficacy of two antimicrobials (Vashe Wound Cleanser and Prontosan Wound Irrigation Solution) against a multispecies bacterial biofilm with or without NPWT in an in vitro wound model., Methods: A mixed multispecies biofilm containing Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pyogenes, and Acinetobacter baumannii was developed and verified by scanning electron microscopy and fluorescent in situ hybridization. The efficacy of Vashe and Prontosan against multispecies biofilm with or without NPWT was evaluated by colony-forming unit (cfu) of each species and total bacterial number, and visually confirmed by live/dead stain and confocal microscopy., Results: Prontosan reduced biofilm cell numbers significantly: 6 instils over 24 h resulting in 3.86 ± 0.14 cfu log10 reduction without NPWT and 4.75 ± 0.13 cfu log10 reduction combined with NPWT (P < 0.01) and 12 instils over 48 h resulting in 5.24 ± 0.11 cfu log10 reduction without NPWT and biofilm eradication with NPWT (P < 0.001). NPWT alone or combined with Vashe failed to reduce multispecies biofilm numbers significantly over 24 or 48 h., Conclusions: Prontosan significantly reduced biofilm cell numbers, with better efficacy over 48 than 24 h, emphasizing the necessity for persistent and robust treatment. NPWT enhanced the effectiveness of Prontosan instillation. However, NPWT alone or combined with Vashe showed limited efficacy and difficulty when combating the multispecies biofilm in vitro., (© The Author(s) 2024. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Purpose: Mice are highly used in retinal research because, like humans, mice have vascularized retinas and choroidal circulation. Although the retinal circulation has been well-characterized in development, its stability during adulthood is less understood. To examine this network, we quantified several key metrics of the trilaminar vasculature., Methods: We used mice (n = 15) with transgenic fluorescent NG2-DsRed (JX: #00824), a vascular-associated label in the retina. One eye per mouse was imaged using confocal microscopy (Nikon A1 Ti2 Eclipse) and traced with ImageJ SNT tools. Using an adaptive optics scanning light ophthalmoscope, additional mice (n = 3) were imaged at single-cell resolution within the living eye to measure the same vasculature., Results: Across mice, we found a stable retinal circulation that formed and maintained a trilaminar stratification throughout early adulthood at all eccentricities. Bridging these layers, microvessels had five distinct anatomical branching patterns. The superficial, intermediate, and deep plexuses increased in density with depth: 16.14 ± 3.61 mm/mm2, 22.14 ± 6.86 mm/mm2, and 31.01 ± 6.24 mm/mm2, respectively. This patterning was not impacted by eccentricity or age (13-61 weeks). Similar metrics were achieved using adaptive optics scanning light ophthalmoscope in vivo with the same analysis pipeline., Conclusions: The mouse retinal vasculature was stable up to 50 weeks of age, providing a robust and extensive baseline dataset with which models of retinal vascular and neural disease may be compared. Vessels connecting the laminae were more complex than previously reported and represented a uniquely vulnerable population due to their relatively low density.

The study of the interactions between biofunctionalized gold nanoclusters (Au NCs) and spermatozoa is highly relevant to evaluate the potential of Au NCs as imaging probes and transfection agents in reproductive biology. In this work, confocal laser scanning microscopy (CLSM) was used to investigate the distribution of Au NCs bioconjugated with peptide (nuclear localisation sequence, NLS) and oligonucleotide (locked nucleic acid, LNA) ligands in bovine spermatozoa. Fluorescence lifetime imaging (FLIM) was employed to detect changes in the NC's chemical environment. We observed a pronounced regio-selective accumulation of the bioconjugates in spermatozoa with high concentration at the equatorial segment. Furthermore, 3D-CLSM showed successful non-endosomal cellular uptake of the conjugates by intact sperm cells and the distribution of the bioconjugates was found to be influenced by the ligand types. Interestingly, the FLIM data showed differences in lifetime depending on membrane integrity. Furthermore, ligand-dependent changes in lifetime between NC bioconjugates carrying peptide and oligonucleotide ligands were found, probably attributed to specific interactions with sperm cell compartments., (© 2024 The Author(s). ChemBioChem published by Wiley-VCH GmbH.)

Objectives: Porphyromonas gulae is a major causative agent of periodontal disease in companion animals that possesses various virulence factors, including fimbriae, lipopolysaccharides, and proteases. P. gulae fimbriae are classified into three genotypes (A, B, and C) based on their nucleotide sequences. Type C fimbrial isolates have been reported to be more virulent than other fimA types, suggesting that different fimA types may aid in the regulation of periodontal pathogenesis. Detailed findings regarding the ability of P. gulae to form biofilms have yet to be reported. Here, we investigated the contributions of fimbrial genotypes in P. gulae biofilm formation., Methods: P. gulae and P. gingivalis biofilms were generated on plates and analyzed using confocal laser microscopy. Additionally, the biofilms formed were assessed by staining with crystal violet. Furthermore, the physical strength of P. gulae biofilms was examined by ultrasonication., Results: Biofilms formed by P. gulae type C were denser than those formed by types A and B. Moreover, the amount of biofilm formed by type C strains was significantly greater than that formed by type A and B strains, which was similar to the biofilms formed by P. gingivalis with type II fimbriae. Additionally, the physical strength of the type C biofilm was significantly greater than that of the other strains., Conclusions: These results suggest that FimA variation may coordinate for biofilm formation. This is the first report on the observation and characterization of P. gulae biofilm formation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved.)

Chronic wounds and chronic ulcers are an increasing problem associated with high health care burden and patient burden. The arrested healing of chronic wounds has, in part, been attributed to the presence of biofilms. Substantial research has documented the presence of biofilms in chronic wounds, and many mechanisms of host-pathogen interactions have been uncovered to explain the arrested healing. However, the paradigm of whether biofilms are only observed in chronic infections was recently challenged when biofilms were also observed in acute infections. Here, we characterize the distribution of bacteria in lower leg wounds with particular emphasis on Pseudomonas aeruginosa and Staphylococcus aureus by confocal laser scanning microscopy combined with PNA-FISH staining and routine culture of bacteria. We show that 40% of wounds contained either P. aeruginosa or S. aureus biofilms and demonstrate the presence of scattered single cells in tissues stained with a universal bacterial PNA-FISH probe. Thus, we demonstrate that chronic wounds do not only harbor bacteria organized in biofilms, but also carry populations of scattered single cells and small cell clusters of only a few bacteria. Our findings may influence diagnostic tools being developed to only target biofilms, where single-cell subpopulations thus may be overlooked and possibly lead to false-negative results., (© 2023 The Authors. APMIS published by John Wiley & Sons Ltd on behalf of Scandinavian Societies for Pathology, Medical Microbiology and Immunology.)

Aim: Natural bioactive products have been tested as alternative antimicrobial agents. This study evaluated the effect of Punica granatum extract (PGE) on oral multispecies biofilms., Methodology: Lyophilized extracts from pomegranate peel were prepared, and the punicalagin content was assessed by ultra-performance liquid chromatography (UPLC). Oral multispecies biofilms from 2 donors were grown on four collagen-coated hydroxyapatite discs. After incubation for 7 days or 3 weeks, the biofilms were exposed to water (control), 2% CHX, 10% PGE, 20% PGE or 30% PGE for 3 min. The proportions of dead bacteria were assessed by the live/dead staining and confocal microscopy. After the analysis, the best PGE concentration (30%) was combined with CHX. The experimental phases were repeated using water, 2% CHX, 30% PGE and 30% PGE + 2% CHX. Five random areas of the biofilm on each disc were scanned, resulting in 20 scanned areas for each group., Results: Regarding the biofilm volume, no differences were found amongst solutions (p = .111). The PGE solution killed bacteria effectively in 1-week, 2-week and 3-week-old-plaque biofilms, ranging from 37 to 55.3%, depending on the PGE concentration. The 30% PGE (a) (p = .0009) had greater antibiofilm effectiveness than 2% CHX (b), which killed bacteria in the 25.2 to 48.7% range. The 10% and 20% PGE had intermediate values (ab), without significant differences from 30% PGE (p = 1.002). Water (c) had the lowest proportion of dead bacteria (p < .00001) in a range of 5 to 6.7% and lower effectiveness in killing bacteria (p < .05). The PGE alone or mixed with 2% CHX had greater anti-biofilm effectiveness than CHX (p < .05). The old plaque biofilms were more resistant than the 7-day-old plaque (p < .05)., Conclusions: The 30% PGE (alone or combined with CHX) exhibited a greater antibiofilm effect on oral multispecies biofilms grown on hydroxyapatite discs than 2% CHX., (© 2024 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Purpose: Landolt ring-shaped epithelial keratopathy is a corneal disease that has only been reported in 11 Japanese patients in 2014. 1 We describe the first case of Landolt ring-shaped epithelial keratopathy in the United States in a patient of European background., Methods: This is a single case report with longitudinal care., Results: A 35-year-old White patient presented with a history of ocular burning, photophobia, and decreased vision. Corneal examination showed bilateral and asymmetric microcystic lesions in a unique Landolt ring (or the letter "C") shape, distributed randomly in the epithelium. Confocal microscopy revealed cellular ballooning and hyperreflective opacities in the basal layer of the corneal epithelium. The patient has had multiple recurrences of her symptoms year-round, each lasting 4 to 8 days. Topical treatment with cyclosporine, steroids, and lubrication resolved her symptoms but without complete resolution of signs on examination., Conclusions: Our patient's clinical signs and symptoms are similar to those described previously in 11 Japanese patients. However, unlike those patients, our patient demonstrates symptomatic response to topical treatment, no seasonal association to her condition, and to date, incomplete resolution of her disease after more than 2 years. This case highlights that Landolt ring-shaped epithelial keratopathy, a novel corneal disease of unclear origin, has relevance outside of the Japanese population., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Plant roots are considered highly efficient soil explorers. As opposed to the push-driven penetration strategy commonly used by many digging organisms, roots penetrate by growing, adding new cells at the tip, and elongating over a well-defined growth zone. However, a comprehensive understanding of the mechanical aspects associated with root penetration is currently lacking. We perform penetration experiments following Arabidopsis thaliana roots growing into an agar gel environment, and a needle of similar dimensions pushed into the same agar. We measure and compare the environmental deformations in both cases by following the displacement of fluorescent beads embedded within the gel, combining confocal microscopy and Digital Volume Correlation (DVC) analysis. We find that deformations are generally smaller for growing roots. To better understand the mechanical differences between the two penetration strategies, we develop a computational model informed by experiments. Simulations show that, compared to push-driven penetration, grow-driven penetration reduces frictional forces and mechanical work, with lower propagation of displacements in the surrounding medium. These findings shed light on the complex interaction of plant roots with their environment, providing a quantitative understanding based on a comparative approach., (© 2024 The Author(s). Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Objectives: Reconstruction of life histories for fossil and living primates draws on rate of enamel layering, termed Retzius periodicity (RP in days) expressed as surface perikymata, during dental crown formation. Disclosure of RP through thin sectioning is destructive; consequently, sample sizes are inadequate to detect the range of RPs present in discrete taxa. We propose an additional method to detect RPs at the population level based on twice-yearly average recurrence of linear enamel hypoplasia (rLEH) in apes shown by prior studies., Materials and Methods: Casts of teeth from orangutans (Pongo pygmaeus) (n = 40) and Lufengpithecus lufengensis (n = 57) from Late Miocene Shihuiba, China, (133 and 138 LEH, respectively) were recorded with scanning electron microscope (SEM) and confocal microscopy to yield perikymata counts between episodes of LEH. Frequency distributions of aggregated perikymata counts between LEH were compared to frequency distribution of tooth-specific ratios of perikymata counts between successive LEH (this latter step removes effects of RP differences within a sample)., Results: Drawing on prior research, two successive intervals between LEH span 1 year on average. Ratios of successive to previous intervals between LEH show that orangutans and Lufengpithecus exhibit two asymmetric intervals centered on 5.3 and 6.7 months, likely reflecting the effect of axial tilt insolation on phenology. Estimated RPs are not unimodal but show a range from 7 to 12 in Lufenpithecus and 8 to 11 in Pongo, comparable to published values., Discussion: Repetitive LEH is sufficiently regular to detect additional RPs which, in the case of Lufengpithecus, have yet to be demonstrated histologically., (© 2024 The Author(s). American Journal of Biological Anthropology published by Wiley Periodicals LLC.)

Objective: To investigate the antibacterial, remineralising, and discolouring effects of copper tetraamine fluoride (CTF) on artificial enamel caries., Method: Human enamel blocks with artificial caries were treated with CTF, silver diamine fluoride (SDF, positive control) and water (negative control) before being challenged with Streptococcus mutans. The morphology, viability, and growth kinetics of biofilm were evaluated using scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), and colony-forming unit (CFU) counting. The lesion depths, mineral loss, micro-hardness, and crystal characteristics were assessed using micro-computed tomography, Knoop Hardness Tester, and X-ray diffraction (XRD), respectively. The discolouring property was assessed by spectrophotometry., Results: SEM showed that bacteria completely covered the enamel surface treated with water, but not the enamel surface treated with CTF and SDF. CLSM showed dead-to-live ratio of biofilm treated with CTF, SDF and water were 0.8 ± 0.1, 0.9 ± 0.2 and 0.5 ± 0.1 (p < 0.001, CTF, SDF>Water). Log 10 CFU values of biofilm treated with CTF, SDF and water were 7.7 ± 0.2, 7.7 ± 0.1 and 8.1 ± 0.1 (p < 0.001, CTF, SDF-3 ) of the CTF, SDF and water groups were 0.4 ± 0.1, 0.4 ± 0.2 and 1.0 ± 0.2 (p < 0.001, CTF, SDFWater). XRD revealed well-crystallised hydroxyapatite in enamel treated with CTF and SDF, but not water. Spectrophotometry showed ΔE values of the CTF, SDF and water groups were 5 ± 3, 54 ± 6 and 6 ± 2 (p < 0.001, CTF, water

Background: In elastic and resistance arteries, an elastin-rich membrane, the Internal Elastic Lamina (IEL), separates the tunica intima from the underlying tunica media. The IEL often appears wrinkled or corrugated in histological images. These corrugations are sometimes ascribed to vessel contraction ex vivo, and to fixation artifacts, and therefore regarded as not physiologically relevant. We examine whether the IEL remains corrugated even under physiological conditions., Methods: The diameters of carotid arteries of anesthetized pigs were measured by ultrasound. The arteries were then excised, inflated within a conical sleeve, fixed, and imaged by confocal microscopy. The conical sleeve allows fixing each artery across a wide range of diameters, which bracket its ultrasound diameter. Thus the study was designed to quantify how corrugations change with diameter for a single artery, and test whether corrugations exist when the fixed artery matches the ultrasound diameter., Results: At diameters below the ultrasound diameter (i.e. when the artery was constricted as compared to ultrasound conditions), the IEL corrugations were found to decrease significantly with increasing diameter, but not fully flatten at the ultrasound diameter. The contour length of the IEL was found to be roughly 10% larger than the circumference of the artery measured by ultrasound. Since ultrasound was conducted with the animal under general anesthesia which induces vasodilation, the physiological diameter is likely to be smaller than the ultrasound diameter, and hence the arteries are likely to have a higher level of corrugation under physiological conditions. For arterial cross sections constricted below the ultrasound diameter, the IEL contour length decreased roughly with the square root of the diameter., Conclusion: The primary conclusions of this study are: a) the IEL is corrugated when the artery is constricted and flattens as the artery diameter increases; b) the IEL is corrugated under physiological conditions and has a contour length at least 10% more than the physiological arterial diameter; and c) the IEL despite being relatively stiffer than the surrounding arterial layers, does not behave like an inextensible membrane., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Background: Atrophic acne scars (AAS) are disfiguring and permanent changes caused by inflammatory acne. Fractional carbon dioxide is a common ablative device used to treat this condition. However, issues such as unclear effectiveness, frequent treatments, and potential side effects exist. In recent years, recombinant human epidermal growth factor (rhEGF) has also been frequently reported for its application in the treatment of acne scars., Objective: To explore the potential synergistic effect of fractional carbon dioxide laser combined with rhEGF in AAS treatment., Methods: We enrolled 15 patients with AAS. They received fractional carbon dioxide laser treatment and were then randomly assigned to receive either rhEGF or a placebo on one side of the face. The procedure was repeated three times, and the results were evaluated using the échelle d'évaluation clinique des cicatrices d'acné (ECCA) score and analyzed using the CBS camera system, 3D analysis (3DMD). Reflectance confocal microscopy (RCM) examination was also conducted., Results: Both sides exhibited significant improvement in the appearance of the acne scars after treatment, as confirmed by the ECCA score, 3DMD data, and CBS texture score. On the rhEGF-treated side, the pore number and epidermal pigment area significantly improved as compared to the control side, whereas no significant differences were observed in the other data. Under RCM, a significant increase in epidermal thickness and appearance of reticular collagen fibers in the dermal layer after treatment was observed., Conclusion: Compared to the sole use of laser, the combination of fractional carbon dioxide laser and rhEGF does not significantly enhance scar therapeutic effects. However, it does shorten the recovery period after laser treatment and improves the pore appearance., (© 2024 The Author(s). Journal of Cosmetic Dermatology published by Wiley Periodicals LLC.)