Μm Intervals Research Articles

Evaluation of Multiple Tissue Levels Frequently Upstages Patients With Clinically Localized Thin Primary Cutaneous Melanoma.

Breslow thickness (BT), ulceration, and microsatellitosis are critical prognostic parameters for cutaneous melanoma staging. These parameters can vary depending on the number of tissue levels examined from individual paraffin blocks. We sought to evaluate all prognostic histopathologic parameters in melanoma for their variations between levels, taken at regular intervals, in a single study. We analyzed 40 consecutive cases of primary cutaneous (nonacral) melanoma through five hematoxylin and eosin sections, taken at 100 μm intervals, for staging and prognostic parameters. Examination of additional levels resulted in (a) an increase in BT in 47.5% (19 out of 40) of cases and (b) detection of ulceration in a further 5% (2/40). This resulted in upstaging for 20% (8 out of 40) of patients (15% because of BT, 2.5% because of ulceration, and 2.5% because of BT and ulceration). The upstaging effect was incremental, with approximately 5% of patients upstaged with each additional 100 μm interval (up to 400 μm). Incipient ulceration and epidermal consumption were infrequent (10% of cases); however, when present, ulceration was subsequently observed in half of cases. We encountered no cases where microsatellitosis was detected at deeper levels. The performance of additional tissue levels is a simple and inexpensive procedure that can improve the accuracy of staging for patients with thin (pT1) primary cutaneous melanomas. It may be pertinent for pathologists to consider additional levels for thin melanomas when a BT measurement is close to a staging threshold (e.g., within 0.1-0.3 mm for pT1a vs. pT1b, or pT1b vs. pT2a), or when incipient ulceration is encountered.

Alternative Strategy to Analyze In Vitro Cell Invasion of 3D Cultures.

Spheroid culture is a 3D model that provides an improved replication of the in vivo microenvironment compared to traditional two-dimensional (2D) cultures. Invasion is a cellular outcome of utmost interest in cancer biology. In this protocol, we have devised an alternative strategy for evaluating cancer cell invasion in vitro, employing heterospheroids comprised of oral squamous cell carcinoma (OSCC), cancer-associated fibroblasts (CAF), and monocytes. These heterospheroids aim to mimic the tumor microenvironment (TME), including two relevant non-neoplastic cell types alongside the cancer cells. Each cell type was labeled with vital fluorescent markers emitting in distinct wavelengths before spheroid formation. Once formed, heterospheroids were seeded onto a layer of human leiomyoma-derived extracellular matrix in the upper compartment of a microporous membrane. Invasion was assessed in the z-axis using confocal microscopy. Digital images were obtained in the corresponding fluorescent channels at 10 µm intervals, covering a depth of 90 µm in the z-axis. Analysis was performed using freeware image software by calculating the integrated fluorescence intensity in each image and fluorescence channel. This approach enables a more dynamic analysis of cell invasion patterns in a multilayered context, as well as the examination of spatial co-localization of different cell types during invasion.

High resolution 3D images of sediment cores as powerful tool for exploring foraminiferal microhabitats

Benthic foraminifera are marine protists largely used as bioindicators and proxies of paleo- environments. Epifaunal species are supposed to live at or above the sediment surface and are therefore used as proxies for bottom water conditions, while infaunal inhabit the sediment column, thus tracing porewater chemistry. Traditional analytical methods based on core slicing, however, have a low resolution that does not allow to precisely characterise the preferential microhabitat(s) of indicator species.In this study we performed microtomographic analyses on an experimental sediment core, to observe the life-position of living foraminifera of two surface-dwelling species Ammonia confertitesta and Haynesina germanica, reported both as epifaunal or shallow infaunal. The images we obtained offered for the first time the possibility to observe each individual in 3D space with a numerical resolution of 13 μm/voxel.The results revealed that the two species are never located above or at the sediment surface and have their preferential microhabitats in a sub-superficial sediment layer constrained in the 0–500 μm interval below the surface. Rapid decrease of abundances below this layer suggests that their microhabitat could be even more specific than previously thought.μCT-scan of sediment cores is also a valuable tool to obtain high-resolution information about foraminiferal ecology. The described method is useful to assess the effective microhabitat of all foraminiferal species that are usually used as proxies for paleorecords, to ensure that the information we can obtain from them is attributable to bottom water or to porewater conditions at a specific sediment depth.

Silk fibroin microgrooved zirconia surfaces improve connective tissue sealing through mediating glycolysis of fibroblasts

The use of zirconia has significantly enhanced the aesthetic outcomes of implant restorations. However, peri-implantitis remains a challenge to long-term functionality of implants. Unlike the perpendicularly arranged collagen fibers in periodontal tissue, those in peri-implant tissue lie parallel to the abutment surface and contain fewer fibroblasts, making them more prone to inflammation. Studies have shown that microgroove structures on implant abutments could improve surrounding soft tissue structure. However, creating precise microgrooves on zirconia without compromising its mechanical integrity is technically challenging. In this study, we applied inkjet printing, an additive manufacturing technique, to create stable silk fibroin microgroove (SFMG) coatings of various dimensions on zirconia substrates. SFMG significantly improved the hydrophilicity of zirconia and showed good physical and chemical stability. The SFMG with 90 μm interval and 10 μm depth was optimal in promoting the proliferation, alignment, and extracellular matrix production of human gingival fibroblasts (HGFs). Moreover, the in vitro results revealed that SFMG stimulated key glycolytic enzyme gene expression in HGFs via the PI3K-AKT-mTOR pathway. Additionally, the in vivo results of histological staining of peri-abutments soft tissue showed that SFMG promoted the vertical alignment of collagen fibers relative to the abutment surface, improving connective tissue sealing around the zirconia abutment. Our results indicated that SFMG on zirconia can enhance HGF proliferation, migration and collagen synthesis by regulating glycolysis though PI3K-AKT-mTor pathway, thereby improving connective tissue sealing.

Salvia tiliifolia leaf extract-based silver nanoparticles for colorimetric detection of Hg(II) in food and environmental samples

In the present research, we provide easy, quick, ultrasensitive, inexpensive, and non-toxic plasmonic sensors using silver nanoparticles (AgNPs). The AgNPs were biosynthesized utilizing Salvia tiliifolia leaf extract and well characterized by various techniques, and then employed to design a colorimetric method enabling Hg(II) sensing. The designed sensor seems very sensitive as well as specific for Hg(II), having limits of detection at 5.0 nM and 0.27 nM through the naked eye and UV–vis detection, sequentially. In addition, the technique showed good precision (intra- and inter-day RSD values were 2.4 % and 4.9 %, respectively) as well as linearity (R2 = 0.9984 over the 0.1–100 µM interval). The actual utilization of the proposed AgNPs-based sensors has been examined in various real samples, including wastewater effluent, lake water, drinking water, soft drinks, mineral water, and canned light tuna. The percentages of recovery of the tested samples varied from 88.0 % to 110 %, demonstrating the method's considerable application. In a nutshell, the method is greener, has tremendous advantages, and has been suggested to detect Hg(II) in foods and environmental samples.

Laser-assisted manipulation of Volta potential pattern on the TC4 surface for improved hBMSCs osteogenesis

The Ti6Al4V (TC4) alloy, a prevalent biomedical material in orthopedics, still faces limitation of the insufficient osseointegration. To improve the bioactivity of TC4, introducing the electric environment onto the TC4 surface may be an effective way in the view of the necessity of endogenous electric microenvironment in bone regeneration. Herein, a Volta potential pattern was engendered on the TC4 surface via parallel laser patterning, so as to promote the osteogenic differentiation of cells. A 15 W laser successfully transformed the original α + β dual phase towards radially distributed lath-like martensite phase in the laser treated region. The atomic lattice distortion between the heterogeneous microstructures of the laser treated and untreated regions leads to a significant Volta potential fluctuation on the TC4 surface. The Volta potential pattern as well as the laser-engraved microgrooves respectively induced mutually orthogonal cell alignments. The hBMSCs osteogenic differentiation was significantly enhanced on the laser treated TC4 surfaces in comparison to the surface without the laser treatment. Moreover, a drastic Volta potential gradient on the TC4 surface (treated with 15 W power and 400 μm interval) resulted in the most pronounced osteogenic differentiation tendency compared to other groups. Modulating the electric environment on the TC4 surface by manipulating the phase transformation may provide an effective way in evoking favorable cell response of bone regeneration, thereby improving the bioactivity of TC4 implant.

Enhancing Wear Resistance in Functionally Graded Metallic Components: Insights from Nanoindentation and Mechanical Analysis.

To manufacture metallic components with high wear resistance, treatments such as nitriding and carburising followed by quenching and tempering (NQT and CQT, respectively) are applied to various types of steel to increase the hardness (H) of the friction surface. However, the wear mechanism of the resulting functionally graded materials has not been fully understood. In this study, specimens of industrial 99.82% pure iron treated with NQT at 913 and 1033 K, and CQT at 1203 K, as well as hot-rolled sheets without heat treatment were examined by performing nanoindentation tests to measure changes in their H, reduced Young's moduli (Er), elastic deformation energies (We), and plastic deformation energies (Wp) along the depth direction. The relationship between Wp/We and the elastic strain resistance (H/Er) can be expressed for all specimens via the equation Wp/We = -1.0 + 0.16 (H/Er)-1. Furthermore, the obtained H/Er av measured at 5 µm intervals based on the specimen profile and wear volume has a good correlation depending to the sliding distance, as confirmed by the results of the ring-on-plate sliding tests conducted for the carbon-treated, nitrogen-treated, and hot-rolled specimens. This study provides a new approach, using H/Er parameters to identify the dominant factors affecting wear resistance at the initial stage of wear that may contribute to the development of wear-resistant materials.

Abstract 2306: A deep learning framework for automated segmentation and analysis of 3D breast cancer spheroid invasion dynamics

Abstract Invasion of neighboring tissues is a cardinal feature of malignancy, notably observed in aggressive cancers like breast cancer, where it can lead to significant morbidity. To explore cancer invasion dynamics, the implementation of a three-dimensional (3D) tumor spheroid invasion assay offers a swift approach to mimicking a tumor micro-region or micro-metastasis. Differential Interference Contrast (DIC) time-lapse imaging was chosen for its fluorescence-free and non-destructive advantages in capturing live spheroid movement. However, the subsequent analysis posed challenges, including out-of-focus cells and a voluminous dataset with diverse microscope planes and time points. Our study introduces a segmentation framework featuring in-focus spatial stacking algorithm and novel training architecture that enabling in-depth automated analysis of 3D spheroid invasion behaviors within a microenvironment. Firstly, the MDA-MB-231 breast cancer spheroids were cultured in a microwell dish and then incorporated into a collagen type I matrix for the 3D spheroid invasion assay. Capturing spheroid invasion dynamics involved acquiring 15 z-plane images (spaced at 20 µm intervals) per timepoint for each spheroid using DIC microscopy at 15-minute intervals over a 28-hour period, employing a 20X objective lens. We refined the Focus Stacking framework into Multiple Slices Blurry Stacking (MSB-Stack) by pre-filtering blurred areas before stacking images. This process produced 6390 slices consolidated into 426 labeled stacked images, highlighting Invasive area, Spheroid core, and Invasive cells post-invasion. We then enhanced Mask R-CNN (Region based-Convolutional Neural Networks) with a weak-to-strong augmentation mechanism, leveraging well-segmented samples from MSB-Stack for training on both fully stacked and blurred images. Blurry Consistency Mask R-CNN (BCMask R-CNN) demonstrates overall improvement across feature extractors, outperforming Mask R-CNN. Using Mean average precision (mAP) as the main metric for instance segmentation and detection, our proposed model achieves 73.0% and 65.2% on mAP @[0.5, 0.95], showcasing robustness. Notably, our best results for mAP @0.5 reach 96.6% and 96.7% on both tasks, underscoring the effectiveness of consistency training in mitigating unclear boundary issues during preprocessing. In conclusion, our proposed method, MSB-Stack, effectively addresses the challenge of unclear boundaries in 3D breast cancer cell DIC microscopy datasets. The integration of weak-to-strong consistency training in our model, BCMask R-CNN, not only mitigates biases but also enhances performance across different backbones, setting the stage for future advancements in cell-tracking tasks and broader migratory analyses. Citation Format: Thi Kim Ngan Ngo, Thanh-Huy Nguyen, Fang-Yu Lin, Ting-Yuan Tu. A deep learning framework for automated segmentation and analysis of 3D breast cancer spheroid invasion dynamics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2306.

In Situ Analysis of the Effect of Ultrasonic Cavitation on Electrochemical Polishing of Additively Manufactured Metal Surfaces

Abstract The effects of a hybrid process that combines ultrasonic cavitation and electrochemical polishing on the electrochemical behavior and the resulting surface characteristics of additively manufactured 316-L stainless steel were investigated. In situ potentiodynamic scans and electrochemical impedance spectroscopy (EIS) were conducted to gain a fundamental understanding of the effect of ultrasonic cavitation on the electrochemical processes involved, considering the influence of electrolyte temperature at 60 and 70 °C. The potentiodynamic scans revealed that increasing the ultrasonic excitation amplitude from 20 to 80 µm at 20 µm intervals and temperature from 60 to 70 °C led to reduced polishing resistance, and elevated passivation current density at equivalent applied potentials, thus leading to an increased polishing rate. These findings are attributed to intensified cavitation near the material surface, which promoted anodic dissolution reactions and accelerated the polishing rate. In situ EIS measurements provided valuable information on the charge transfer resistance and double-layer capacitance and their influence on the hybrid process. Specifically, higher ultrasonic amplitudes and elevated temperatures contributed to enhanced electrical double-layer formation and ion adsorption, resulting in a faster rate of polishing, indicating the efficacy of the hybrid process. These findings enhance our understanding of the complex interactions between ultrasonic cavitation and electrochemical dissolution processes that occur during ultrasonic cavitation-assisted electrochemical polishing. The research provides valuable insights for optimizing the process and its potential application in the post-processing of metal additive manufactured parts.

Analysis of the 3.2–3.3 μm Interstellar Absorption Feature on Three Milky Way Sightlines

We report new analyses of spectra of the 3.2–3.3 μm absorption feature observed in the diffuse interstellar medium toward three Milky Way sources: 2MASS J17470898 − 2829561 (2M1747) and the Quintuplet Cluster, both located in the Galactic center, and Cygnus OB2-12. The 3.2–3.3 μm interval coincides with the CH-stretching region for compact polycyclic aromatic hydrocarbons (PAHs). We focus on the 2M1747 spectrum. Its published optical depth spectrum contains residual telluric transmission features, which arise from the 0.06 difference in mean airmasses between the observations of the source and its telluric standard star. We corrected the published spectrum by adding the airmass residual optical depth spectrum. The corrected spectrum is well fit by a superposition of four Gaussians. The absorption spectra of the other two sources were also fit by four Gaussians, with similar central wavelengths, widths, and relative peak opacities. We associate the three longer wavelength Gaussians covering the 3.23–3.31 μm interval with compact PAHs in positive, neutral, and negative charge states. We identify the shortest-wavelength Gaussian, near 3.21 μm, with irregularly shaped PAHs. Constraints imposed by spectral smoothness on the corrected 2M1747 spectrum, augmented by a PAH cluster formation model for post-asymptotic giant branch stars, suggests that >99% of the PAHs in the diffuse interstellar medium reside in small clusters. This study supports the PAH hypothesis, and it suggests that a family of primarily compact PAHs with a C66H20 (circumvalene) parent is consistent with the observed mid-infrared and ultraviolet interstellar absorption spectrum.

An Angle‐Sensitive Microcolumn‐Based Capacitive Shear Force Sensor for Robot Grasping

AbstractShear force sensors play an indispensable role in tactile perception for robot manipulation tasks. However, recent advancements in shear force sensors have been hindered by issues such as direction sensitivity and integration limitations. This paper proposes a microcolumn array dielectric layer produced using photolithography technology that enables tunability of sensor sensitivity and detection range by adjusting the aspect ratio and interval of the microstructures. Meanwhile, the impact of five constant normal force couplings on the sensitivity of shear force perception is investigated. The structure array with a 1:2 aspect ratio and 600 µm interval demonstrates an ultrahigh sensitivity of 6.189 N−1 and outstanding linearity (R2 = 0.9873) within the range up to 0.1 N. The sensor exhibits low hysteresis and robust stability over 3000 cycles. Additionally, it exhibits remarkable anisotropic direction sensitivity, enabling accurate positioning within a quarter‐circle angle. An intentionally designed orthogonal array is employed to extend the shear angle range up to 360°. Owing to the high performance of the sensor, it is further integrated onto a gripper to facilitate the grasping operation and effectively capture delicate movements. The experimental outcomes highlight that the designed sensor holds promise for applications in robotic applications and electronic skin domains.

Comparative evaluation of slide scanners, scan settings, and cytopreparations for digital urine cytology

BackgroundAcquiring well-focused digital images of cytology slides with scanners can be challenging due to the 3-dimensional nature of the slides. This study evaluates performances of whole-slide images (WSIs) obtained from 2 different cytopreparations by 2 distinct scanners with 3 focus modes. MethodsFourteen urine specimens were collected from patients with urothelial carcinoma. Each specimen was equally divided into 2 portions, prepared with Cytospin and ThinPrep methods and scanned for WSIs using Leica (Aperio AT2) and Hamamatsu (NanoZoomer S360) scanners, respectively. The scan settings included 3 focus modes (default, semi-auto, and manual) for single-layer scanning, along with a manual focus mode for 21 Z-layers scanning. Performance metrics were evaluated including scanning success rate, artificial intelligence (AI) algorithm-inferred atypical cell numbers and coverage rate (atypical cell numbers in single or multiple Z-layers divided by the total atypical cell numbers in 21 Z-layers), scanning time, and image file size. ResultsThe default mode had scanning success rates of 85.7% or 92.9%, depending on the scanner used. The semi-auto mode increased success to 92.9% or 100%, and manual even further to 100%. However, these changes did not affect the standardized median atypical cell numbers and coverage rates. The selection of scanners, cytopreparations, and Z-stacking influenced standardized median atypical cell numbers and coverage rates, scanning times, and image file sizes. DiscussionBoth scanners showed satisfactory scanning. We recommend using semi-auto or manual focus modes to achieve a scanning success rate of up to 100%. Additionally, a minimum of 9-layer Z-stacking at 1 μm intervals is required to cover 80% of atypical cells. These advanced focus methods do not impact the number of atypical cells or their coverage rate. While Z-stacking enhances the AI algorithm's inferred quantity and coverage rates of atypical cells, it simultaneously results in longer scanning times and larger image file sizes.

Strength and Microscopic Mechanism of Cement-Fly Ash-Slag-Desulfurization Gypsum Solidified Mica Schist Weathered Soil.

Mica schist weathered soil possesses a number of poor engineering characteristics, which make it difficult to use as a subgrade material for resource utilization. Therefore, in this study, a new type of curing agent, CFSD (cement-fly ash-slag-desulfurized gypsum), is proposed for this soil. The effects of different curing agent dosages, age of preservation, and confining pressure on the stress-strain curves were analyzed via the uniaxial compression test and triaxial compression test, while the micromorphological characteristics of cured soil were analyzed via X-ray diffraction analysis and the SEM test combined with Image J software. In this paper, we also establish a microscopic mechanism model to determine how curing agents increase the strength of mica schists. The results reveal that the compressive strength of solidified soil increases rapidly within 28 days; the CFSD dosage of 4% at 7 d increased by 103.23% by 28 d. After 28 d, the trend of compressive strength growth was flat. The CFSD dosage of 4% at 7 d increased by 128.34% by 90 d; with the increase in the dosage, the curve transformed from flat to steep. These results suggest that the CFSD dosage is positively correlated with the damage strain and damage bias stress of solidified soil. The curves for the strain softening type with a 4% dosage as the initial effective confining pressure increased from 50 kPa to 300 kPa; the failure stress and failure strain increased by 202.09% and 90.85%, respectively. With the increase in curing agent dosage and maintenance age, the pore size of 2~5 μm, >5 μm interval decreased from 56.46% to 27.92%, the porosity decreased from 12.51% to 4.6%, and the hydrate produced by the curing agent cemented and filled up the pore space between the loose particles of the soil body. Thus, the large pore space became microporous, and the pore structure densification was greatly improved.

Compare of optic coherence tomography parameters in recreational synthetic tetrahydrocannabinol use and healthy control

Purpose To evaluate retinal thickness (RT), retinal nerve fiber layer thickness (RNFLT), and choroidal thickness (CT) changes in synthetic cannabinoid (SC) users. Methods This prospective study evaluated the RT, RNFLT, and CT values of 56 SC users and 58 healthy controls. The individuals using SCs were referred to us by our hospital’s forensic medicine department. Retinal and choroidal images were obtained using spectral-domain optical coherence tomography (OCT). Measurements (one subfoveal, three temporals, three nasal) were taken at 500 μm intervals up to 1500 μm using the caliper system. Only the right eye was used for subsequent analysis. Results Mean ages were 27.7 ± 5.7 years in the SC-user group and 25.4 ± 6.7 in the control group. Subfoveal Global RNFLT was in the SCs group 102.3 ± 10.5 μm and 105.6 ± 20.2 μm in the control group (p = 0.271). Subfoveal CT was in the SC group mean of 316.1 ± 100.2 μm and in the control group mean 346.4 ± 81.8 μm (p = 0.065). RT, T500 (283.3 ± 36.7 μm, 296.6 ± 20.5 μm, p = 0.011) and N1500 (355.1 ± 14.3 μm, 349.3 ± 18.1 μm, p = 0.049) were significantly higher in the SC group than in the control group, respectively. Conclusion Analysis of OCT findings of individuals who had been using SC for more than one year revealed no statistically significant difference between RNFLT and CT, although N1500 was significantly higher in RT. Further studies in the field of OCT are important to explore the pathology of SC

Centrosome fragmentation and malposition is common in the first cell division of equine ICSI embryos

In vitro production of embryos via Intra-Cytoplasmic-Sperm-Injection (ICSI) has become popular in the horse industry. Although improving, the blastocyst yield in clinical practice remains sub-optimal (∼20%: Stout. JEVS 2020; 89:103011). The high incidence of developmental arrest of ICSI embryos is suggested a result of mitotic errors during the first zygotic cell divisions. A better understanding of the cause of these errors could help develop strategies to prevent mitotic errors in equine in vitro produced (IVP) embryos. In somatic cells, the centrosomes, composed of two centrioles surrounded by pericentriolar material are the major microtubule-organizing centers and play essential roles in spindle assembly and chromosome segregation. Mammalian oocytes lack centrosomes and although two centrioles are re-introduced by the spermatozoon at fertilization, centrosomes make only a minor contribution to zygotic spindle assembly (Schneider et al., J.Cell. Biol. 2021; 220(11):e202010106). Although not essential for spindle assembly, the role of the centrosomes in ensuring fidelity of chromosome segregation during zygotic cell division is unclear. The aim of this study was to evaluate the position and number of centrosomes in horse ICSI zygotes during the first mitotic division. Cumulus-Oocyte-Complexes (COCs) were obtained from Warmblood mares by Ovum-Pick-Up, maintained in H-SOF at room temperature overnight and then matured in vitro, fertilized by ICSI and cultured as described previously (Ducheyne et al. Reprod. Fertil. Dev. 2019; 31(12):1830). After 24h, injected oocytes were fixedand immunostained for alpha-tubulin, acetylated tubulin, (peri-)centrosome material (NEDD1) and chromatin (DAPI). For each zygote, confocal microscopy was performed using a Leica SPE II microscope and a 0.17µm interval z stack generated. Of 326 COCs collected, 177 reached MII and were injected and cultured further. Of the 177, 136 zygotes were fixed and imaged, 13 were at the two pronucleus, 11 at prophase/prometaphase, 14 at metaphase, 11 at anaphase or telophase, 54 at the two cell stage and 33 were either degenerated or poorly stained. During the two pronucleus and prophase/prometaphase stages, centrosomes mainly localized at the interface between the two pronuclei. In 56% (n=14/25) of zygotes in pro-metaphase and metaphase, independent spindles formed around each of the two parental pronuclei. The majority of dual spindles were closely adjacent and synchronous. Moreover, in 64.2% (n=9/14) of the metaphase zygotes, the (peri-)centrosomes were either malpositioned (e.g. not at the poles of the spindle) or fragmented. Fragmented or multiple centrosomes are known to predispose to abnormal chromosome-microtubule attachment and aberrant chromosome segregation. We therefore hypothesize that malposition and fragmentation of the centrosomes contributes to mitotic errors and developmental arrest of equine ICSI embryos. Acknowledgements to CAPES, Brazil, for financial support (Code 88882.433332/2019-01 and 88887.467895/2019-00)

Study of the human hippocampal formation: a method for histological and magnetic resonance correlation in perinatal cases.

Little information is available on the magnetic resonance imaging (MRI) determination of the hippocampal formation (HF) during the perinatal period. However, this exploration is increasingly used, which requires defining visible HF landmarks on MRI images, validated through histological analysis. This study aims to provide a protocol to identify HF landmarks on MRI images, followed by histological validation through serial sections of the temporal lobe of the samples examined, to assess the longitudinal extent of the hippocampus during the perinatal period. We examined ex vivo MRI images from nine infant control brain samples. Histological validation of the hippocampal formation MRI images was obtained through serial sectioning and examination of Nissl-stained sections at 250μm intervals along the entire length of the hippocampal formation. Up to six landmarks were identified both in MRI images and the serial histological sections. Proceeding in an anterior to posterior (rostrocaudal) direction, these were as follows: 1) the limen insulae (fronto-temporal junction); 2) the beginning of the amygdaloid complex; 3) the beginning of the lateral ventricle; 4) the caudal limit of the uncus, indicated by the start of the lateral geniculate nucleus (at the level of the gyrus intralimbicus); 5) the end of the lateral geniculate nucleus (beginning of the pulvinar); and 6) the beginning of the fornix. After histological validation of each of these landmarks, the full longitudinal length of the hippocampal formation and distances between landmarks were calculated. No statistically significant differences were found in total length or between landmarks. While the HF is anatomically organized at birth, its annotation is particularly challenging to perform. The histological validation of HF landmarks allows a better understanding of MRI images. The proposed protocol could be useful to assess MRI hippocampal quantification in children and possible variations due to different neurological diseases.

Hyperreflective dots in the avascular outer retina in relapsing-remitting multiple sclerosis

Hyperreflective granular elements with a transient presence in the retina can be detected non-invasively by optical coherence tomography (OCT). Such foci or dots may represent aggregates of activated microglia. However, in multiple sclerosis an increased number of hyperreflective foci has so far not been demonstrated in the intrinsically hyporeflective and avascular outer nuclear layer of the retina where there are no fixed elements in healthy eyes. Therefore, the present study intended to investigate the presence of hyperreflective foci in the outer nuclear layer in patients with relapsing- remitting multiple sclerosis (RRMS) by using a high-resolution OCT scanning protocol. This cross-sectional exploratory study examined 88 eyes in 44 RRMS patients and 106 eyes in 53 age- and sex-matched healthy subjects. None of the patients had any sign of retinal disease. All patients and healthy subjects each underwent one session of spectral domain OCT imaging. A total of 23,200 B-scans extracted from 8×8 mm blocks of linear B-scans at 60 μm intervals were analysed for hyperreflective foci in the outer nuclear layer of the retina. Analyses were made of the total block scan and a circular 6-mm diameter fovea-centered field in each eye. Multivariate logistic regression analysis was used to assess associations between parameters. Hyperreflective foci were observed in 31 out of 44 (70.5 %) multiple sclerosis patients compared to 1 out of 53 (1.8%) healthy subjects (p < 0.0001). From analyses of the total block scans, the median number of hyperreflective foci in the outer nuclear layer was 1 (range 0-13) in patients and 0 (range 0-2) in healthy subjects (p < 0.0001). In total, 66.2% of all hyperreflective foci were located within 6 mm of the center of the macula. There was no detectable association between the presence of hyperreflective foci and retinal nerve fiber layer or ganglion cell layer thickness. Hyperreflective granular foci in the avascular outer nuclear layer of the retina seen by OCT were almost completely absent in healthy subjects, whereas they were found, albeit at low density, in the majority of patients with RRMS. Hyperreflective foci can be repeatedly examined by non-invasive means and without pupil dilation, which opens a new field of investigation of infiltrating elements in an unmyelinated part of the central nervous system.

초정밀 레이저를 이용한 연꽃잎 표면 자연 모사 연구

In a pilot natural super-hydrophobic surfaces study, a super-hydrophobic surface was made by coating, etching, laser ablation, chemical vapor deposition and lithography. In this study, cone-shaped periodic micro and nano-structures were constructed on a silica surface with femtosecond and picosecond laser, and the period of micro-structures between cone shape patterns was increased with 10 μm intervals. The contact angle and image of the super-hydrophobic surface were analysed and the cone (Aspect-ratio 1.27) shape model with micro-protrusion structure similar to the surface of the lotus leaf was made to measure the contact angle. To analyse the differences in the contact angles between the cone shapes and heights of the micro-protrusion, different samples with cone (Aspect-ratio 1.27), sphere (Aspect-ratio 1.00) shapes were made through laser micro-machining technology. To mimick the natural lotus leaves, the optimum condition was a cone shape. Samples of PDMS with different shapes and mixed micro/nano-structures were fabricated with a PDMS mold insert. The largest contact angle was measured at 170.42° which is similar to the contact angle of the lotus leaf. This mold insert could be used repeatedly. The molding process is advantageous for large areas and mass production.

Edi Mod Spectral Domain Optical Coherence Tomography Evaluation of The Choroid in Retinitis Pigmentosa: A Case-Control Study

Objective: The evaluation of choroidal thickness measurements became possible after the developments in optical coherence tomography (OCT) technology. This study aimed to evaluate choroidal features in Retinitis Pigmentosa subjects with EDI (Enhanced depth imaging) mode spectral domain optical coherence tomography (SD-OCT). Material and Method: A hundred and three eyes of 54 RP subjects underwent scanning with EDI-OCT for central retinal and choroidal thickness measurements and were compared with 40 healthy controls. Submacular choroidal thickness measurements were obtained beneath the fovea and at 500 µm intervals for 2.5 mm nasal and temporal to the center of the fovea. Results: Mean subfoveal choroidal thickness measurements were 253.2±74.9 µm in RP patients and 336.2±91.7 µm in control (p&lt;0.0001). There was no correlation between subfoveal choroidal thickness and visual acuity in RP patients. The choroid had an irregular shape in 85 % of RP patients. The thickest point of the choroid was not subfoveal as in healthy eyes, and excessive nasal thinning of the choroid was observed in 93 % of RP patients. Conclusion: Submacular choroidal thickness was significantly reduced but did not correlate with the visual acuity in RP patients.

Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold.

In vitro studies on adherent cells require a process of passage to dissociate the cells from the culture substrate using enzymes or other chemical agents to maintain cellular activity. However, these proteolytic enzymes have a negative influence on the viability and phenotype of cells. The mesenchymal stem cell (MSC)-like cell line, C3H10T1/2, adhered, migrated, and proliferated to the same extent on newly designed microporous titanium (Ti) membrane and conventional culture dish, and spontaneous transfer to another substrate without enzymatic or chemical dissociation was achieved. The present study pierced a 10 μm-thick pure Ti sheet with 25 μm square holes at 75 μm intervals to create a dense porous structure with biomimetic topography. The pathway of machined holes allowed the cells to access both sides of the membrane frequently. In a culture with Ti membranes stacked above- and below-seeded cells, cell migration between the neighboring membranes was confirmed using the through-holes of the membrane and contact between the membranes as migration routes. Furthermore, the cells on each membrane migrated onto the conventional culture vessel. Therefore, a cell culture system with enzyme-free passaging was developed.