Additive manufacturing (AM) is a promising new technology that is having a very fast growth from home workshops to high-tech cutting-edge factories. As any manufacturing technique, adequate metrology services are needed to assure the quality of items manufactured by AM. One of the most widely used instruments to measure the characteristics of surfaces manufactured with AM is the confocal microscope. In this paper, authors present a whole calibration procedure for confocal microscopes designed to be implemented preferably in workshops or industrial environments rather than in research and development departments. Because of that, it is as simple as possible. The procedure is designed without forgetting any of the key aspects that need to be taken into account and based on classical reference material standards. These standards can be easily found in industrial dimensional laboratories and easily calibrated in accredited calibration laboratories.

Breast cancer remains a significant health concern worldwide, necessitating early detection and treatment. The rapid appearance or a change in the shape of skin lesion on the breast or just of the nipple-areola complex(NAC), especially if it is unilateral needs to be evaluated. Several primary or secondary tumours such as inflammatory skin lesions can affect this body area. Ultrasound, mammography, or magnetic resonance imaging (MRI) are the most used non-invasive diagnostic techniques for breast cancer, but in recent years reflectance confocal microscopy (RCM) and optical coherence tomography have emerged as additional tools to diagnose cases characterized by cutaneous changes.The introduction of new techniques such as reflectance confocal microscopy (RCM), may open new perspectives in the non-invasive diagnosis of mammary paget disease (MPD). RCM is a non-invasive diagnostic technique that allows high-resolution images of the skin at microscopic level in real time. It is non-invasive, painless and repeatable, making it a valuable tool in the early diagnosis and monitoring of skin diseases. We describe an atypical clinical presentation of mammary paget disease early diagnosed by reflectance confocal microscopy evaluation.

Rapid microscopic analysis of tissue is an essential diagnostic tool in oncological surgery. The gold standard for intraoperative histological tissue evaluation are frozen sections. However, frozen sections are prone to a variety of artefacts and require skilled staff and specialized lab equipment. A potential method for rapid intraoperative tissue evaluation that does not require fixation, freezing nor sectioning of the tissue, is ex vivo fluorescence confocal microscopy (FCM). The visualization of the structurally important extracellular matrix (ECM) in conventional ex vivo FCM lags behind the standards of conventional histology. The objective of this study was to find a stain that will improve the depiction of the ECM to resemble FFPE H&E sections as closely as possible.

Background: MRI-guided prostate biopsies from visible tumor-specific lesions (TBx) can be used to diagnose clinically significant carcinomas (csPCa) requiring treatment more selectively than conventional systematic biopsies (SBx). Ex vivo fluorescence confocal microscopy (FCM) is a novel technique that can be used to examine TBx prior to conventional histologic workup. Methods: TBx from 150 patients were examined with FCM on the day of collection. Preliminary findings were reported within 2 hours of collection. The results were statistically compared with the final histology. Results: 27/40 (68%) of the csPCa were already recognized in the intraday FCM in accordance with the results of conventional histology. Even non-significant carcinomas (cisPCa) of the intermediate and high-risk groups (PSA > 10 or 20 ng/ml) according to conventional risk stratifications were reliably detectable. In contrast, small foci of cisPCa were often not detected or were difficult to distinguish from reactive changes. Conclusion: The rapid reporting of preliminary FCM findings helps reduce the psychological stress on patients an can improve the clinical management of csPCa. Additional SBx can be avoided in individual cases, leading to lower rates of complications and scaring in the future surgical area. Additional staging examinations can be arranged without losing time. FCM represents a promising basis for future AI-based diagnostic algorithms.

Hyper duplex stainless steels, HDSS, are a new alloy group of duplex stainless steels with the highest corrosion resistance and mechanical properties among the existing modern stainless steel. Due to the addition of extra high content of alloying elements, e.g. Cr, Ni, Mo, etc., the crystallization behavior of the δ-ferrite from the liquid is of vital importance for controlling the steel properties. In this work, the effect of cooling rate (i.e. 4°C/min and 150 °C/min) on the growth process of δ-ferrite in S33207 during the solidification process was investigated using high-temperature confocal scanning laser microscopy (HT-CLSM) in combination with electron microscopes and thermodynamic calculation. The results showed that the solidification mode of S33207 steel was a ferrite-austenite type (FA mode). L→δ transformation occurred at a certain degree of supercooling, and merging occurred during the growth of δ phase dendrites. Similar microstructures were observed after solidification under these two different cooling rates. The variation of area fraction of δ ferrite at the free surface and temperature as well as time during solidification of S33207 steels were calculated at different cooling rates of 4 °C/min and 150 °C/min, respectively. The current HT-CLSM study provides in-situ experimental evidence to confirm that a low cooling rate favored the growth of primary δ-ferrite, whereas a high cooling rate favored the nucleation of primary δ-ferrite during the solidification process. The post-microstructure as well as composition evolution is also briefly investigated. This work shed light on the real time insights for the crystallization behavior of hyper duplex stainless steels during solidification, which aims to contribute to the process control of manufacturing of this steel grade.

Both the use of plastic and its decomposition products lead to the distribution of plastic all over the earth and finally to the uptake by all kind of living beings including humans. Still, it is widely unknown what risks harbor the widespread uptake of plastics for human health, espe-cially regarding contributing factors like size, shape and surface composition. We assessed the uptake of polymethylmethacrylate (PMMA)-nano- and microbeads for HEK293-, A549- and MRC5 cells. Via confocal microscopy, we localized multiple PMMA-beads inside the cytosol of cells. Uptake of PMMA-beads did not alter cell growth and cell division, implying no short-term toxicity towards human cells. Further, we used a cAMP response element binding protein (CREB)-mediated reporter assay to assess whether internalized PMMA-nanobeads alter cell-signaling pathways. In contrast to the in vitro transcription, where the addition of PMMA-nanobeads abolished the transcription, no changes regarding CREB-mediated cell sig-naling are given in HEK293-cells. Our data led to the assumption that PMMA-nano-and mi-crobeads are internalized via endocytosis and end up as lysosomes within the cells cytosol. Therefore, we concluded differences regarding the surface composition of the PMMA-nanobeads mainly affect its potential to alter cell signaling. These findings emphasize the key role the sur-face composition plays regarding microplastic and its risks for human health.

Tumour spheroids are fast becoming commonplace in basic cancer research and drug development. Obtaining high-quality data relating to the inner structure of spheroids is important for analysis, yet existing techniques often use equipment that is not commonly available, are expensive, laborious, cause significant size distortion, or are limited to relatively small spheroids. We present a high-throughput method of mounting, clearing, and imaging tumour spheroids that causes minimal size distortion. Spheroids are mounted in an agarose gel to prevent movement, cleared using a solution prepared from commonly available materials, and imaged using confocal microscopy. We find that our method yields high quality two- and three-dimensional images that provide information about the inner structure of spheroids.

The purpose of this study was to describe ocular surface and anterior eye segment findings in various types of ichthyoses. Methods: This was a single-center prospective observational study. The study group consisted of five patients (P1-P5) aged 13–66 years. Multimodal imaging was performed, including slit-lamp examination, swept-source optical coherence tomography (SS-OCT), and in vivo confocal microscopy (IVCM). Results: All patients were diagnosed with moderate to severe dry eye disease (DED). Corneas showed a significant pattern of irregularity, with a significant difference between the corneal thickness at the apex (CAT) and corneal thinnest thickness (CTT), ranging from 5 to 375 µm. Three patients were diagnosed with ectasia patterns based on SS-OCT. All patients showed abnormalities in at least one Fourier index parameter for at least one eye at 3 or 6 mm, on keratometric, anterior, or posterior analyses. IVCM examination revealed changes in all corneal layers. Conclusions: By combining the results of multimodal im-aging, we were able to detect preclinical abnormalities, distinguish characteristic changes com-mon to ichthyosis, and reveal the depth and characteristics of corneal abnormalities. Therefore, patients with ichthyosis should be examined for DED and ectatic disorders early in clinical prac-tice.

Gliomas are one of the most aggressive and treatment-resistant types of human cancer. One of the most promising field in gliomas cancer therapy is identification and evaluation of anticancer properties of compounds found in plants i.a. naringenin (N) and 8-prenylnaringenin (8PN). The prenyl group seem to be crucial to the anticancer activity of flavones, which may lead to enhanced cell membrane targeting and thus increased intracellular activity. Unfortunately, 8PN content in hop cones is from 10 to 100 times lower compared to other flavonoids i.e. xanthohumol. In this study we used a simple method for the synthesis of 8PN from isoxanthohumol, via O-demethylation with high, 97% of the isolated yield. Cellular accumulation and cytotoxicity of naringenin and 8-prenylnaringenin in normal (BJ) and cancer cells (U-118 MG) were also examined. Obtained data indicated that 8-prenylnaringenin exhibited higher toxicity against used cell lines than naringenin and both flavones inhibited stronger glioblastoma U-118 MG cells than normal fibroblasts. The anticancer properties of 8PN correlated with its significantly greater (37%), accumulation in glioblastoma cells than in normal fibroblasts. Additionally, naringenin indicated higher selectivity for glioblastoma as it was over 6 times more toxic for cancer than normal cells. Our results provide evidence that examined prenylated and non-prenylated flavanones have different biological activity against normal and cancer cell lines and this phenomenon may be useful in clinical practice to construct new, anticancer drugs for glioblastoma.

In a large part of ecological niches, bacterial species are united in communities and develop together in the form of heterogeneous structures called biofilms, which are formed on surfaces of different nature and nature with the participation and as a result of the interaction of two or more bacterial species. The main objective of the present work is to study the effects of pH-value in the culture medium on the biofilm development and architecture of biofilms of the bacterium (B. subtilis) during their interactions with Escherichia coli K-12 1655 strain. Effects of pH-value in the culture medium on bacterial biofilm formation and the mechanisms were analyzed by the crystal violet staining method combined with cultivated microbial analysis, and confocal laser scanning microscopy. In the presence of phosphate at pH-value of pH 6,0 , the bacterial growth rates of single-species biofilms of Bacillus subtilis strains and biofilms as result of their interaction with Escherichia coli strains were similar and determines the mutualism between two strains in the structure of biofilms, formed by the participation of B.subtilis 170 and E.coli K-12 1655, B.subtilis 168 and E.coli K-12 1655 strains. Considering that static growth in a biofilm is a highly relevant growing state for many soil bacteria, more research on phosphate acquisition and pH homeostasis by model soil bacteria in static cultures is required to draw a more comprehensive picture of the role of biofilm production in phosphate use efficiency and pH homeostasis.

Silk fibroin is a well-known biopolymer used in several applications in which the interaction with biological tissue is required. In fact, fibroin is extremely versatile and can be shaped to form several constructs useful in tissue engineering applications. Confocal imaging is usually per-formed to test the cells behaviour on the construct and in this context the fibroin autofluorescence is regarded as a problem. In addition, the autofluorescence is not intense enough to provide useful morphological images. In fact, to control study the constructs morphology other techniques are used (i.e. SEM, Micro-CT). In this work we propose a method based on the fluorescence energy transfer (FRET) to suppress the fibroin autofluorescence moving it to higher wavelength accessible to the confocal microscopy for a direct imaging.

Immunofluorescent methods in vitro have been widely used to characterize the distribution and extent of colocalization of molecules located in various intracellular compartments. These are key processes to better understand the role of those molecules in physiological and pathological conditions.In the brain, interactions of astrocytes with neurons are fundamental for the establishment and proper functioning of neuronal circuits. Among these, the phagocytosis of synapses that need to be removed is a process known as synaptic pruning. When this process is dysregulated, it might lead to the development of several brain disorders. Here, we describe the adaptation of a method to characterize the astrocyte-mediated synaptic phagocytosis and evaluate astrocytic (dys)functions in healthy cells and cells derived from an animal model of major depressive disorder (MDD).

The present paper explored the possibility of discriminating between different malignant degrees of Chondrosarcoma (CS), a diffused bone tumor, and Enchondroma (EC), its benign version, through an approach based on the coupling between Confocal Raman Microscopy (CRM) and Machine Learning (ML) techniques. Recently, Raman Spectroscopy has proven to be a powerful tool for a complete grading of CS by distinguishing between grade 1, 2 and 3, and to distinguish CS from EC. In this paper, we tested some models, belonging either to ML or to the sub-type called Deep Learning (DL), showing excellent classification performances, especially the DL algorithms, with classification accuracy approaching to 100%, making the models promising for future implementations of Raman spectroscopy when applied to oncology diagnosis. In turn, we highlighted how some proper ML models result in worse classification performances but better resolution of specific chemical target compounds, possible candidate to become malignant markers, with relevant implication for a correct diagnosis.

Purpose: To evaluate the relationship between corneal nerve structure and function in a veteran population. Methods: 83 veterans (mean age: 55 ± 5 years) seen at the Miami Veterans Affairs (VA) eye clinic were included in the study. Each individual filled out questionnaires to evaluate ocular symptoms (5-Item Dry Eye Questionnaire, DEQ5, Ocular Surface Disease Index, OSDI) and ocular pain (Numerical Rating Scale, NRS, Neuropathic Pain Symptom Inventory modified for the Eye, NPSI-Eye). Individuals also underwent an ocular surface examination that captured functional nerve tests including corneal sensation, corneal staining, and Schirmer test for tear production. Corneal sub-basal nerve analysis was conducted using in vivo confocal microscopy (IVCM) images with corneal nerve density, length, area, width, and fractal dimension captured. IVCM and functional corneal metrics from the right eye were examined using correlational and linear regression analysis. Results: Most corneal structural metrics were not related to functional metrics, except for weak correlations between various IVCM metrics and tear production. In addition, corneal nerve fiber area was positively related to corneal sensation (r = 0.3, p = 0.01). On linear regression analyses, only corneal fractal dimension remained significantly related to tear production (β = -0.26, p = 0.02) and only corneal nerve fiber area remained significantly related to corneal sensation (β = 0.3, p = 0.01). Conclusions: Most corneal nerve structural metrics did not relate to functional metrics in our veteran population, apart from a few weak correlations between structural metrics and tear production. This highlights caution in using corneal nerve anatomy to predict corneal function.

Djeya1 (RKLAFRYRRIKELYNSYR), is a very effective cell penetrating peptide (CPP) which mimics the α5 helix of the highly conserved Eya domain (ED) of eyes absent (Eya) proteins. The objective of this study was to bioengineer analogues of Djeya1 that, following effective translocation into planarian tissues, would reduce the ability of neoblasts (totipotent stem cells) and their progeny to regenerate the anterior pole in decapitated S. mediterranea. As a strategy to increase the propensity for helix formation, molecular bioengineering of Djeya1 was achieved by the mono-substitution of the helicogenic aminoisobutyric acid (Aib) at three species variable sites: 10,13,16. CD analyses indicted that Djeya1 is highly helical and that Aib-substitution had little influence on secondary structure. Aib-substituted Djeya1 analogues are highly efficient CPPs, devoid of influence upon cell viability or proliferation. All three peptides increase the migration of PC-3 cells which express high concentrations of Eya. Two peptides, [Aib13]Djeya1 and [Aib16]Djeya1, are bioportides which delay planarian head regeneration. As neoblast are the only cell population capable of division in planaria, these data indicate that bioportide technologies could be utilised to directly manipulate other stem cells in situ, thus negating any requirement for genetic manipulation.

Fused deposition modeling (FDM) is a rather new technology in the production of personalized dosage forms. Melting and printing of polymer–active pharmaceutical ingredient (API) – mixtures can be used to produce oral dosage forms with different dosage as well as release behavior. This process is utilized to increase the bioavailability of pharmaceutically relevant active ingredients that are poorly soluble in physiological medium by transforming them into solid amorphous dispersions (ASD). Release from such ASDs is expected to be faster and higher compared to the raw materials and thus enhance bioavailability. Printing directly from powder while forming ASDs was realized. Different techniques to change release patterns as well as a non-destructive way for determination of API distribution were shown. By measuring the melt viscosities of mixtures printed a rheological model for the printer used is proposed.

Tissue clearing methods combined with confocal microscopy have been widely used for studying developmental biology. In plants, ClearSee is a reliable clearing method that is applicable to a wide range of tissues and is suitable for gene expression analysis using fluorescent reporters, but its application to the Arabidopsis thaliana embryo, a model system to study morphogenesis and pattern formation, has not been described in the original literature. Here we describe a ClearSee-based clearing protocol, which is suitable for obtaining 3D images of Arabidopsis thaliana embryos. The method consists of embryo dissection, fixation, washing, clearing, and cell wall staining, and enables high quality 3D imaging of embryo morphology and expression of a fluorescent reporter with the cellular resolution.

Self-assembling films typically used for colloidal lithography have been applied to pine wood substrates to change the surface wettability. Therefore, monodisperse polystyrene (PS) spheres have been deposited onto a rough pine wood substrate via dip coating. The resulting PS sphere film resembled a polycrystalline FCC-like structure with typical domain sizes of 5 – 15 single spheres. This self-assembled coating was further functionalized via an O2 plasma. This plasma treatment strongly influenced the particle sizes in the outermost layer, and hydroxyl as well as carbonyl groups were introduced to the PS spheres’ surfaces, thus generating a superhydrophilic behaviour.

Supramolecular solvent (SUPRAS) has received increasing interest as an innovative, efficient, and green solvent for the effective extraction and separation of bioactive compounds from natural resources. However, studies on the use of SUPRAS for the extraction of phenolic components from plants, which are highly valued in food products due to their excellent antioxidant capacity, are still lacking. The present study developed a green ultrasound-assisted and efficient SU-PRAS method to simultaneously determine three phenolic acids in Prunella vulgaris by high-performance liquid chromatography (HPLC) and the experimental parameters were optimized in detail. The efficiency and antioxidant properties of the phenolic compounds obtained by different extraction methods were also compared. Under the optimal conditions, the extraction efficiency of SUPRAS, prepared with octanoic acid reverse micelles dispersed in ethanol-water, was substantially higher than conventional organic reagents. Moreover, this SUPRAS method also showed more significant antioxidant capacities. The confocal laser scanning microscopy (CLSM) images presented the spherical droplet structure of SUPRAS (A well-defined circle), which coincided with the position of the fluorescence position of phenolic acids. The phenolic acids were encapsulated in the vesicles by the SUPRAS droplets, which indicated the efficient extraction capacities of SUPRAS for the target compounds. Finally, combined with the CLSM method, the mechanism of the proposed method was further clarified, and the superior extraction performance of SUPRAS was theoretically demonstrated through molecular dynamics simulations. In comparison with conventional methods, the higher extraction efficiency of SUPRAS can be illustrated by the larger solvent contact surface area, more types of hydrogen bonds between extractants and supramolecular solvents, and stronger and more stable interaction forces. The results of the theoretical studies are in rational agreement with the experimental outcome.

Surfaces of additively manufactured metal parts from powder-based processes typically show powder particle agglomerations and other features, resulting in high surface roughness. Proper characterization of those surfaces is necessary in order to assess part quality with respect to coatability, mechanical performance or corrosion resistance for use in aerospace, automotive, medical and more industrial applications. Optical surface texture measurement allows for collection of areal surface data, while the established contact stylus method only captures line profile data. When applying optical methods for surface topography measurements, proper data acquisition and post-processing in order to assess surface texture may be complex. A number of variables can be adjusted, such as different measurement settings, approaches to outlier removal, evaluated area size or form removal. This work shows the influence of selected z-range prior to measurement and the influence of choosing pre-defined outlier removal settings in MountainsMap 9.2 on selected ISO 25178-2:2022 parameters calculated from data obtained from confocal microscopy for as-built Ti6Al4V from laser powder bed fusion. The aim is to show the influence of variation in measurement and post-processing on calculated surface texture parameters and stress the importance of proper documentation in order to achieve reproducibility of data for quality management.

Glaucomatous optic neuropathy, a major cause of blindness, is characterized by the loss of retinal gan-glion cells (RGCs) and degeneration of their axons. Mitochondria are deeply involved in maintaining RGCs and their axons. Therefore, lots of attempts have been made to develop diagnostic tools and therapies tar-geting mitochondria. Recently, we reported that mitochondria are uniformly distributed in unmyelinated axons of RGCs, possibly owing to the ATP gradient. Thus, using transgenic mice expressing yellow fluo-rescent protein targeting mitochondria exclusively in RGCs within the retina, we assessed the alteration of mitochondrial distributions induced by optic nerve crush (ONC) via in vitro flat-mount retinal sections and in vivo fundus images captured through the cornea with a confocal scanning ophthalmoscope. We observed that the mitochondrial distribution in unmyelinated axons of RGCs that survived the ONC remained uni-form, although their density increased. Furthermore, via in vitro analysis, we discovered that the mitochon-drial size is attenuated following ONC. These results suggest that ONC induces mitochondrial fission without disrupting the uniform mitochondrial distribution. This mechanism may help to prevent axonal degeneration and apoptosis. The in vivo visualization system of axonal mitochondria in RGCs may be ap-plicable in the diagnosis of progression of optic neuropathy.

Transplant of human cancer cells into zebrafish larvae has emerged as a useful methodology in cancer research. Zebrafish have very low husbandry costs, are amenable to large-scale drug screening, and are unmatched for optical clarity in live animal imaging. However, there is currently no consensus on the ideal methods for xenograft of human cancer cells into zebrafish. Here, we have examined the effects of transplant site and housing temperature on both zebrafish larvae and human cancer cells using survival analyses, metabolomic approaches, and in vivo imaging. Our data show that while zebrafish larvae can adapt to the ideal conditions for mammalian cells, human cancer cells are highly sensitive to both temperature change and transplant site. Human cells housed in slightly cooler than physiologic temperatures had a significantly altered metabolism that resulted in changes in growth, survival, and response to chemotherapy. Cancer cells xenografted into the yolk of the larvae also had reduced proliferation and drug response compared to those xenografted into the soma, in part due to the differences in metabolites available at these sites. In total, temperature and transplant site can have profound effects on xenografted cells. Standardization of zebrafish xenograft methods will enhance data reproducibility between individual laboratories.

Polylactic Acid (PLA) is an organic polymer commonly used in fused deposition (FDM) printing and biomedical scaffolding that is biocompatible and immunologically inert. However, variations in source material quality and chemistry make it necessary to characterize the filament and determine potential changes in chemistry occurring as a result of the FDM process. We used several spectroscopic techniques, including laser confocal microscopy, Fourier-Transform Infrared (FTIR) spectroscopy and photoacousitc FTIR spectroscopy, Raman spectroscopy, and X-ray photoelectron Spectroscopy (XPS) in order to characterize both the bulk and surface chemistry of the source material and printed samples. Scanning Electron Microscopy (SEM) and Differential Scanning Calorimetry (DSC) were used to characterize morphology, crystallinity, and the glass transition temperature following printing. Analysis revealed calcium carbonate-based additives which were reacted with organic ligands and potentially trace metal impurities, both before and following printing. These additives became concentrated in voids in the printed structure. This finding is important for biomedical applications as carbonate will impact subsequent cell growth on printed tissue scaffolds. Results of chemical analysis also provided evidence of the hygroscopic nature of the source material and oxidation of the printed surface, and SEM imaging revealed micro and sub-micron scale roughness that will also impact potential applications.

This study aimed to evaluate the impact of Candida albicans on subgingival biofilm formation on dental implant surfaces. Scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) were used to compare biofilm structure and microbial biomass in the presence and absence of the fungus after periods of 24, 48, and 72 h. Quantitative polymerase chain reaction (qPCR) was used to quantify the number of viable and total micro-organisms for each of the biofilm-forming strains. A general linear model was applied to compare CLSM and qPCR results between control and test conditions. The biofilm developed with C. albicans at 72 h had a higher bacterial biomass, with a significantly higher cell viability (p < 0.05). After both 48 and 72 h of incubation, in the presence of C. albicans, there was a significant increase in counts of Fusobacterium nucleatum and Porphyromonas gingivalis and in the cell viability of Streptococcus oralis, Aggregatibacter actinomycetemcomitans, F. nucleatum, and P. gingivalis. Using a dynamic in vitro multispecies biofilm model, C. albicans exacerbated the development of the biofilm grown on dental implant surfaces, significantly increasing the number and cell viability of periodontal bacteria.

Infectious keratitis (IK) is among the top 5 leading causes of blindness globally. Early diagnosis is key to guide an appropriate therapy to avoid complications such as vision impairment and blindness. Culture of corneal scrapes is the initial diagnostic test to grow and identify the causal organism. Alternative diagnostic tools include imaging such as with optical coherence tomography (OCT) and in vivo confocal microscopy (IVCM). OCT’s advantage is its ability to accurately determine the depth and extent of the corneal ulceration, infiltrates and haze; therefore, characterizing the severity and progression of the infection. However, it is not a preferred choice in the diagnostic tool package for infectious keratitis. IVCM is a great aid in the diagnosis of fungal and Acanthamoeba keratitis with overall sensitivities of 66-74% and 80-100%, and specificity of 78-100% and 84-100%, respectively. Recently the use of deep learning (DL) models in IK has been shown to aid diagnosis via image recognition. Most of the studies that have developed DL models to diagnose the different types of IK have utilised photographs from digital camaras, slit-lamp images, or IVCM images. Some studies have used extremely efficient single DL algorithms to train their models and others used ensemble approaches. This technology is likely to assist in the diagnosis of IK in some years. Further work is needed to examine and validate the clinical performance of the DL models in the real-world setting and to evaluate whether this technology improves patient clinical outcomes. The scope of this review was to provide a recent update on the diagnostic imaging tools in IK

The term neuroinflammation defines the reactions of astrocytes and microglia to alterations in homeostasis in the diseased central nervous system (CNS), the exacerbation of which contributes to the neurodegenerative effects of Alzheimer's disease (AD). Local environmental conditions, such as the presence of proinflammatory molecules, mechanical properties of the extracellular matrix (ECM) and local cell-cell interactions, are determinants of glial cell phenotypes. In AD, the load of the cytotoxic/proinflammatory amyloid β peptide (Aβ) is a microenvironmental component increasingly growing in the CNS, imposing time-evolving challenges on resident cells. This study aimed at investigating the temporal and spatial variations of the effects produced by this process on astrocytes and microglia, either directly or by interfering in their interactions. Ex vivo confocal analyses of hippocampal sections from the mouse model TgCRND8 at different ages have shown that overproduction of Aβ peptide induced early and time persistent disassembly of functional astroglia syncytium and promoted a senile phenotype of reactive microglia, hindering Aβ clearance. In the late stages of disease, these patterns were altered in the presence of Aβ-plaques, surrounded by typically reactive astrocytes and microglia. Morphofunctional characterization of peri-plaque gliosis revealed a direct contribution of astrocytes in plaque buildup that might result in shielding Aβ-peptide cytotoxicity and, as a side effect, in exacerbating neuroinflammation.

This paper presents the experimental implementations of the mathematical models and algorithms developed in Part I. Two experiments are carried out. The first experiment aims at the determinations of the correction coefficients of the mathematical model. The dot grid target is measured and the measurement data are processed by our developed and validated algorithms introduced in Part I. The values of the coefficients are indicated and analysed. Uncertainties are evaluated with implementation of the Monte Carlo method. The second experiment measures a different area of the dot grid target. The measurement results are corrected according to the coefficients determined in the first experiment. The mean residual between the measured points and their corresponding certified values reduced 29.6% after the correction. The sum of squared errors reduced 47.7%. The methods and the algorithms for raw data processing, such as data partition, fittings of dots’ centres, K-means clustering, etc., are the same for both two experiments. The experimental results demonstrate that our method for the correction of the errors produced by the movement of lateral stage of confocal microscope is meaningful and practicable.

Extramammary Paget disease (EMPD) is an uncommon adenocarcinoma of apocrine gland-rich areas, presenting significant diagnostic challenges due to its nonspecific clinical appearance and frequent misidentification as benign, inflammatory skin conditions. Traditional diagnostic methods such as biopsy are invasive and uncomfortable, often required repeatedly due to high recurrence rates. Dermoscopy and non-invasive imaging techniques have been used but provide limited diagnostic accuracy due to their constraints in depth penetration and resolution. Recent advancements in imaging technologies, such as line-field confocal optical coherence tomography (LC-OCT), show promise in enhancing diagnostic precision while minimizing invasive procedures. LC-OCT merges high-resolution imaging with deep penetration capabilities, capturing detailed horizontal and vertical skin images akin to histopathology. This study evaluated the diagnostic performance of LC-OCT in detecting EMPD and its recurrence in 17 clinically suspicious anogenital regions, belonging to 6 patients. Data were collected prospectively at patient bedside by an LC-OCT expert with poor training for EMPD; and, then, reviewed retrospectively by an independent LC-OCT expert with adequate training for EMPD and no concerns about time. The prospective examination yielded 64.7%-accuracy, 71.4%-sensitivity, and 33.3%-specificity. The retrospective analysis achieved 94.1%-accuracy, 100%-sensitivity, and 66.7%-specificity, with the only false positive case being a difficult-to-diagnose concomitant presentation of a lichen sclerosus et atrophicus. Despite the need for specialized training, our results suggest that LC-OCT represents a valuable tool for accurately identifying EMPD and improve its management by reducing unnecessary biopsies. Further studies are needed to standardize its clinical application.

The effect of sonication on the molecular characteristics of poliacrylic acid (Carbopol® Ultrez 10) and its rheological behavior in aqueous dispersions and microgels containing 0.25 wt. % of the polymer was analyzed in this work by rheometry, weight-average molecular weight (Mw) measurements via static light scattering (SLS), Fourier transform infrared (FTIR) spectroscopy and confocal microscopy. For this, the precursor dispersion and the microgels were sonicated in a commercial ultrasound bath at constant power and different times. We observed a softening of the microgel microstructure consisting of a systematic decrease in its shear modulus, yield stress and viscosity with increasing sonication time, while their overall Herschel-Bulkley (H-B) behavior was preserved. SLS measurements evidenced a reduction of Mw of polyacrylic acid with sonication time. Separately, FTIR measurements indicate that sonication produces scission in the C-C links of the Carbopol® backbone, which results in chains with the same chemistry but lower molecular weight. Finally, confocal microscopy measurements revealed a concomitant diminution of the size of the microsponge domains with sonication time, which is reflected in a softer microstructure resulting from reduction of the molecular weight of polyacrylic acid. The present results indicate that both the microstructure and the rheological behavior of Carbopol® microgels, in particular, and complex fluids in general, may be manipulated or tailored by high-power ultrasonication.

Skin cancer is one of the most prevalent cancers worldwide, with an increasing incidence. Skin cancer is typically classified as melanoma or non-melanoma skin cancer. Melanoma, despite being much less common than basal or squamous cell carcinomas, is the most deadly, with nearly 8,300 Americans expected to die from melanoma each year. Biopsies are currently the gold standard in diagnosing melanoma; however, they can be invasive and expensive and inaccessible to lower-income individuals. Currently, suspicious lesions are triaged with image-based technologies, such as dermatoscopy and confocal microscopy. While these techniques are useful, there is wide inter-user variability and minimal training for dermatology residents on how to properly use these devices. The use of artificial intelligence (AI)-based technologies in dermatology has emerged in recent years to assist in the diagnosis of melanoma that may be more accessible to all patients and more accurate than current methods of screening. This review explores the current status of the application of AI-based algorithms in the detection of melanoma, underscoring its potential to aid dermatologists in clinical practice. We will specifically focus on AI’s application in clinical imaging, dermoscopic evaluation, algorithms that can distinguish melanoma from non-melanoma skin cancers, and in-vivo skin imaging devices.

Age-related macular degeneration (AMD) is a leading cause of blindness worldwide. In late-stage AMD, geographic atrophy (GA) of dry AMD or choroidal neovascularization (CNV) of neovascular AMD results in macular atrophy (MA), leading to significant visual loss. Despite the development of innovative therapies, there are currently no established effective treatments for MA. As a result, early detection of MA is critical in identifying later central macular involvement throughout time. Accurate and early diagnosis is achieved through a combination of clinical examination and imaging techniques. Our review of the literature depicted advances in retinal imaging to identify biomarkers of progression and risk factors for late AMD. Imaging methods like fundus autofluorescence (FAF), near-infrared reflectance (NIR), widefield imaging, optical coherence tomography (OCT), multicolor confocal scanning laser ophthalmoscopy (cSLO), and optical coherence tomography angiography (OCTA) can be used to detect and monitor the progression of retinal atrophy. Multifocal electroretinogram (ERG) and microperimetry are methods for quantifying visual function to map disease progression. The evolving diverse imaging modalities optimize detection of pathology anatomy and measurement of visual function; they may also contribute to the understanding of the underlying mechanistic pathways, particularly the underlying MA changes in late AMD.

Metastasis in oncological diseases remains one of the main reasons for negative prognosis regarding treatment. Any new data on the biophysical and biochemical characteristics of circulating metastatic cells will help to develop the concept of antimetastatic therapy. In this study we found a number of differences in spectroscopic and morphological features of circulating metastatic cells. FTIR and Raman spectra of cultivated by adhesive and de-adhesive methods (last as a model for metastatic cells) have showed spectroscopic features, namely in FTIR in the region of CH - stretching vibrations associated with structural rearrangements in the cell membrane, as well as changes in the intensity and position of the PO2- group vibrations bands correlated with a proliferative activity. The spectral features in the regions of OH stretching and Amid I vibrations and other spectral markers of the metastatic cells under different cultivation conditions were derived. Raman spectra showed redistribution of amino acid Tyr/Trp ratio and in Tyr duplet intensity in the region of 500-900 cm-1 , as well different glycogen level in different cells. The spectroscopic markers are in the accordance with biochemical data. CARS and confocal optical microscopy were applied for determination a state of the cells and F-actin expression level that turned out to be higher for adhesive cells in comparison with de-adhesive cells. Shape and morphological property of the cells drastically differs. The correlation of vibrational markers with biochemical data and cytofluorometric method were discussed.

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc) is a major disease affecting banana production around the globe. To study the movement of Foc in planta and investigate potential physical mechanisms of deterrence, the roots of a set of banana cultivars were challenged with a GFP-tagged subtropical race 4 strain of Foc. These plants included susceptible and resistant varieties including the plantain 'FHIA25' (AAB), dessert hybrids 'Lady Finger' (AAB), 'FHIA02' (AAAA), as well as the industry preferred Cavendish line, 'Williams' (AAA), and its somaclone derivative 'GCTCV119' (AAA). GFP localisation in planta confirmed that Foc can infect all four cultivars. Its movement through roots to the rhizome and to the aerial parts of the plants was evident. Tyloses formation in the root vasculature was also detected by GFP auto-fluorescence and further confirmed using scanning electron microscopy. While tyloses formation did not prevent colonisation and infection, their formation restricted the fungus within the xylem vessels, slowing its proliferation. This mechanism may be an important factor in providing plant host tolerance to Foc but it doesn't provide complete immunity. Fluorescently tagged Foc is a useful tool for understanding the infection process of the Fusarium wilt disease of banana and aiding in the integrated management of this important disease.