Adjacent Sections Research Articles

Isotope Encoded chemical Imaging Identifies Amyloid Plaque Age Dependent Structural Maturation, Synaptic Loss, and Increased Toxicity.

It is of critical importance to our understanding of Alzheimer's disease (AD) pathology to determine how key pathological factors are interconnected and implicated in nerve cell death, clinical symptoms, and disease progression. The formation of extracellular beta-amyloid (Aβ) plaques is the major pathological hallmark of AD and Aβ has been suggested to be a critical inducer of AD, driving disease pathogenesis. Exactly how Aβ plaque formation begins and how ongoing plaque deposition proceeds and initiates subsequent neurotoxic mechanisms is not well understood. The primary aim of our research is to elucidate the biochemical processes underlying early Aβ plaque formation in brain tissue. We recently introduced a chemical imaging paradigm based on mass spectrometry imaging (MSI) and metabolic isotope labelling to follow stable isotope labelling kinetics (iSILK) in vivo to track the in vivo build-up and deposition of Aβ. Herein, knock-in Aβ mouse models ( App NL-F ) that develop Aβ pathology gradually are metabolically labeled with stable isotopes. This chemical imaging approach timestamps amyloid plaques during the period of initial deposition allowing the fate of aggregating Aβ species from before and during the earliest events of plaque pathology through plaque maturation to be tracked. To identify the molecular and cellular response to plaque maturation, we integrated iSILK with single plaque transcriptomics performed on adjacent tissue sections. This enabled changes in gene expression to be tracked as a function of plaque age (as encoded in the Aβ peptide isotopologue pattern) distinct from changes due to the chronological age or pathological severity. This approach identified that plaque age correlates negatively with gene expression patterns associated with synaptic function as early as in 10-month-old animals but persists into 18 months. Finally, we integrated hyperspectral confocal microscopy into our multiomic approach to image amyloid structural isomers, revealing a positive correlation between plaque age and amyloid structural maturity. This analysis identified three categories of plaques, each with a distinct impact on the surrounding microenvironment. Here, we identified that older, more compact plaques were associated with the most significant synapse loss and toxicity. These data show how isotope-encoded MS imaging can be used to delineate Aβ toxicity dynamics in vivo. Moreover, we show for the first time a functional integration of dynamic MSI, structural plaque imaging and whole genome-wide spatial transcriptomics at the single plaque level. This multiomic approach offers an unprecedented combination of temporal and spatial resolution enabling a description of the earliest events of precipitating amyloid pathology and how Aβ modulates synaptotoxic mechanisms.

Tricolored Bat (Perimyotis subflavus) microsite use throughout hibernation

Abstract White-nose syndrome (WNS) has caused dramatic population declines in several bat species, including the Tricolored Bat (Perimyotis subflavus). Several studies have documented bats using colder roosting temperatures after infection; however, this strategy may have costs such as increased freezing risks or greater predation risks and it is unknown when during hibernation bats begin to utilize these colder temperatures. Our aim was to examine Tricolored Bat roost locations in a WNS-positive site in relation to roost microclimate and other environmental conditions throughout the hibernation season. We conducted monthly censuses of tricolored bats across 2 hibernation seasons (November to March 2020 to 2021 and October to March 2021 to 2022) in a WNS-positive hibernaculum in northwestern South Carolina and recorded skin and adjacent wall temperature, tunnel section, and distance from the entrance for each bat species. We continuously measured hibernacula temperature and relative humidity during both hibernation seasons. Most bats roosted in the back part of the tunnel where temperatures were warmer and more stable, and vapor pressure deficit (VPD) and human disturbance were low. However, >20% of bats roosted in the front, where roost temperatures were significantly colder and VPD was higher but more variable; human disturbance was also higher in this section. The proportion of bats in each tunnel section did not vary among months and we did not find evidence of significant movement to the front section of the tunnel as hibernation progressed based on marked bats; however, bats in the front section roosted higher on the wall suggesting that they may be avoiding human disturbance or predators. Our results support the notion that no optimum hibernation temperature exists for tricolored bats and that high VPD and disturbance are likely important factors driving microsite use. Protection of Tricolored Bat hibernacula that offer a range of microclimates or a network of sites in close proximity that offer different microclimates may be helpful for recovery of this species.

7662 Mass Spectrometry Imaging Reveals Region-, Age-, And Sex-Specific Effects Of Lipopolysaccharide Administration On Glucocorticoids In Mouse Lymphoid Organs

Abstract Disclosure: M. Salehzadeh: None. S. Khan: None. R. Andrew: None. K.K. Soma: None. Glucocorticoids (GCs) are steroids that are critical for immune function. Immune organs, such as the thymus and spleen, locally produce GCs (i.e., immunosteroids). However, GC concentrations in the thymus and spleen at the cellular level, and how these levels are affected by stress, remain largely unknown. We used state-of-the-art mass spectrometry imaging (MSI) to elucidate the spatial distribution of GCs in the neonatal and adult mouse thymus and spleen 1) at baseline and 2) after an acute stressor. We used MSI to measure 11-deoxycorticosterone (DOC), corticosterone, and 11-dehydrocorticosterone (DHC) in different regions of the thymus (medulla/cortex) and spleen (white/red pulp) after an immunological challenge. We administered lipopolysaccharide (LPS; i.p.) at low (50μg/kg) and high (400μg/kg) doses or saline (vehicle) to male and female C57BL/6J mice at post-natal day (PND) 5 (neonatal) and 90 (adult) (n=6/sex/group/age). 4hr after treatment, we collected thymus and spleen on liquid nitrogen. Cryosections (10μm) were subject to MSI following Girard-T reagent derivatization and α-cyano-4-hydroxycinnamic acid matrix application. Matrix assisted laser desorption/ionisation (MALDI) was used for sampling, coupled to Fourier-transform ion cyclotron mass spectrometry (100μm resolution; positive mode; Bruker 12T SolariX). Adjacent sections were taken to determine histological zones by H&E staining. Preliminary results indicate that LPS increased DOC, corticosterone, and DHC levels by 2- to 9-fold in lymphoid organs at both ages. There was a dose-dependent increase in thymus and spleen GC levels at PND5, but not PND90. At PND5, GC levels were similarly distributed across each tissue in both sexes. In contrast, at PND90, thymic corticosterone levels were greater in males than females in the medulla and cortex. Interestingly, in PND90 thymus, the medulla had greater DHC levels than the cortex only after LPS treatments. In PND90 spleen, GC levels significantly increased after LPS, yet there was no significant difference in GC distribution between sexes or red and white pulp. These results suggest that GC production is uniform across neonatal lymphoid organs. However, in adulthood, the thymus exhibits region-specific GC metabolism. Our results expand knowledge of steroid intracrinology in thymus and spleen and potential effects of environmental stressors (e.g., infectious diseases, toxins) on immune GC responses. Further, our novel MSI method for GC detection in murine lymphoid organs can help inform future techniques in pharmacokinetics for localized thymic and splenic targeting of steroids and steroid modulators. Presentation: 6/3/2024

Influence of lithodynamics of coastal area on the safety of railway operation on the Tuapse-Adler section of the North Caucasus Railway

The present paper considers the railroad bed on the Tuapse-Adler section of the North Caucasus Railway (Black Sea coast) in terms of its protection from wave scouring. Aim. To evaluate the influence of coastal lithodynamics on the safety of the railway operation on the section under consideration. The paper describes lithodynamic processes in the sea coastal zone and demonstrates the influence of coastal protection structures produced on lithodynamics in general and on adjacent sections, as exemplified by a specific section of the coast (1907–1910 km). The study involves a retrospective analysis of the step-by-step construction of trapping structures, and sea groins without full-fledged dumping of beach-forming material into the inter-groin compartments. The investigation and analysis of satellite images of 1907–1910 km coast within the Vodopadny-Lazarevskoye section indicate that lithodynamic processes remained unconsidered during the intensive construction of bank protection structures, which led to the beach erosion and the emergency condition of the seawalls along 1909–1910 km section. The paper provides recommendations for considering lithodynamic processes in the construction of railway protection structures, thereby increasing the safety of the railway operation.

Переходные участки бесстыкового пути, примыкающие к стрелочным переводам

Purpose: consider the problem of interaction between turnouts and adjacent sections of a seamless railway track. Methods: statistical data analysis. Results: an analysis of the results of domestic and foreign studies has been carried out, which show that in such areas there is a jump with subsequent extrusion of material of less rigidity and an increase in the slope of the elastic unevenness, i. e., an undesirable phenomenon occurs — a shock wave. The impact force depends on the mass of the rolling stock and the speed of movement, the geometric evenness of the rail track and the chosen approaches to designing the transition zone as a whole. Jump and mainly occurs due to compression and pushing out of structural layers and materials of lower rigidity in the under-rail area. The impact force depends on the mass of the rolling stock and the speed of movement, the geometric evenness of the rail track and the chosen approaches to designing the transition zone as a whole. Jump and mainly occurs due to compression and pushing out of structural layers and materials of lower rigidity in the under-rail area. Shock waves caused by uneven rigidity of sections have negative consequences (the appearance of deviations in level, subsidence, distortions, increased intensity of rail wear, the appearance of rail defects), which increase the cost of maintenance of the railway track. An analysis of the accumulation of residual deformations has been carried out, which shows the systematic formation of geometric irregularities in adjacent areas in the turnout area. Practical importance: from the point of view of the design of the transition zone, the expected advantage will be the differentiation of structural materials and elements and their influence on achieving the optimal gradation of railway track rigidity in the vertical and horizontal (transverse, longitudinal directions), selection of the optimal ratio of the mechanical properties of individual elements of the superstructure structure. It should also be noted that it is necessary to conduct an additional feasibility study of the feasibility of constructing transition sections in the area where the track and switches meet, taking into account the operational features of individual railway lines.

B-350 The application of MALDI-TOF imaging to biomarker discovery in a mouse model of papillomavirus associated tumorigenesis

Abstract Background Human papillomavirus (HPV) can cause infection and dysplasia in cutaneous epithelium of immunodeficient patients. An animal model has been developed to study HPV-associated tumorigenesis using Nude mouse (Foxn1nu) and mouse papillomavirus (MmuPV1). This study applies MALDI-TOF imaging to compare the expression pattern of small molecules between viral-infected dysplastic mouse skin and mock-infected control tissue. Three sets of molecules showing distinct patterns are identified in correlation with the upregulation of an enzyme in related biosynthetic pathway. Methods Nude mice were wounded on tail and inoculated with MmPV1 suspension (1×109 viral DNA genomes per infected site) or PBS 24 hours later for treatment and control group, respectively. Tail skins were harvested 4 weeks post-infection and processed for cryosectioning. Tissue sections were analyzed using timsTOF flex (Bruker) under negative ion mode with 9-aminoacridine as the matrix. Viral infection (MmuPV1-L1 antigen) and enzyme expression (3'-phosphoadenosine 5'-phosphosulfate synthetase 1, PAPSS1) were assessed by immunofluorescence on adjacent sections. Results Three peaks demonstrated distinct distribution patterns in mouse skin. One peak at m/Z = 465 was identified as cholesterol sulfate (CS) by tandem MS and comparing with the standard. CS was enriched in keratinocytes, hair follicles, and blood vessels in normal skins, and it increased in dysplastic skins with enhanced angiogenesis. The combined peaks at m/Z = 327 & 329 were tentatively identified as docosahexaenoic acid (DHA) and docosapentaenoic acid (DPA). These long chain polyunsaturated fatty acids (LC-PUFAs) were present in the dermis as foci partially co-localized with CS, and they also increased in virus-infected skins. The peak at m/Z = 426 may represent a combination of ADP and APS. Although these two isobars cannot be separated by tandem MS, APS was revealed by the presence of sulfur isotopes at expected natural abundance (34S and 32S). They were present at a low level in the epidermis of normal skins but significantly increased after virus infection. APS and ADP were closely associated with the proliferating cells, and the signal intensity also increased with the degree of tissue dysplasia. As a single factor, APS & ADP demonstrated a fair differential power for MmuPV1-induced dysplastic skin and normal control (AUC of ROC curve = 0.75). The bifunctional enzyme PAPSS1 catalyzes the conversion of ATP to 3’-phosphonato-5’-adenyl sulfate (PAPS) via APS as an intermediate. Interestingly, immunofluorescence showed the expression of PAPSS1 also increased significantly with the severity of tissue dysplasia and MmuPV1 infection. Conclusions MALDI-TOF imaging reveals three sets of molecules with distinct patterns in mouse skin, and their levels are increased during MmuPV1-induced epithelium dysplasia. As a precursor of the common sulfate donor (PAPS), increased APS and ADP in the epidermis of virus-infected skins indicate an upregulation of sulfation pathway and adenine metabolism. This finding is supported by enhanced expression of enzyme PAPSS1 and an increased sulfation reaction product, CS. In addition, DHA and DPA are also increased in the dermis of virus-infected skins, implying a disturbance in LC-PUFA metabolism. This study demonstrates combined use of MALDI-TOF imaging and immunostaining in the search of novel biomarkers of HPV-associated tumorigenesis.

Immunocytochemistry assessment of vocal fold regeneration after cell-based implant in rabbits.

Cell-based outer vocal fold replacement (COVR) offers a potential treatment for severe vocal fold scarring or cancer reconstruction. Previous work in rabbits using human adipose-derived stem cells (ASC) in fibrin suggested that a hybrid structure emerged within 2 months, containing both implanted and host cells. This project uses immunocytochemistry to better define the phenotypic fate of implanted cells and features of the extracellular environment. Immunocytochemistry was performed on sections collected from rabbits 2 months after COVR implantation or scar surgery. Cellular targets included human leukocyte antigen (HLA), CD31, and smooth muscle actin (SMA). HLA was present in all implanted sections and was used to identify human cells. In adjacent sections, HLA-positive cells were identified expressing CD31. SMA was not identified in the same cells as HLA. These markers were also present in injured vocal folds not receiving COVR. SMA protein content did not differ according to treatment. Implanted human ASC persist in rabbit vocal folds. Some appear to express CD31, an endothelial marker. Smooth muscle actin, a marker of myofibroblast phenotype, was present in all sections regardless of treatment, and was not identified in hASC. Host cells also infiltrate the structure, producing a hybrid host-graft vocal fold.

The geodynamics of plume-influenced mid-ocean ridges: insights from the Foundation Segment of the Pacific-Antarctic Ridge

The intersection of the Foundation Plume with the Pacific-Antarctic Ridge is a key location in global geodynamics where a mantle plume is approached by and interacting with a fast-spreading mid-ocean ridge. Here, we discuss a comprehensive major and trace element and Sr-Nd-Pb isotope dataset of new and existing samples from the young Foundation Seamount Chain (<5 Ma) and adjacent section of the Pacific-Antarctic Ridge. We use the geochemistry of axial, off-axis and intraplate lavas to map the spatial extent of plume dispersal underneath the ridge as well as the internal zonation of the upwelling plume. We show that the unusual length, increased crustal thickness and occurrence of silicic rocks on the axis of the Foundation Segment are the direct result of plume being tapped by the axial melting zone. We demonstrate that the plume is not homogeneous but shows a HIMU-like (high time-integrated 238U/204Pb) OIB (Ocean Island Basalt) component characterized by 206Pb/204Pb of up to 20.5 in its center and a more EM1-like (Enriched Mantle one) OIB component characterized by low U/Pb and 206Pb/204Pb but high Rb/Nb and 87Sr/86Sr towards its edges. Plume entrainment leads to a high magma supply rate that fosters the formation of silicic rocks and triggers the lengthening of the segment over time. However, plume dispersal is not symmetric as the geochemical tracers for the OIB component are extending <100 km northwards but >300 km southwards. We relate this to the current plate tectonic framework in which the obliquity between the migrating ridge and the absolute plate motions induces a sub-axial asthenospheric flow that preferentially channels plume material southwards.

Design and mechanical properties analysis of a novel CFRP tendons anchorage structure

To address the challenges of larger size, stress concentration at the loading end, and insufficient anchoring performance at the free end of traditional bonding-type anchors for Carbon Fiber Reinforced Polymer (CFRP) tendons, a novel anchor featuring a variable angle multi-stage cone is proposed. This solution is derived by analyzing the notch effect at the loading end caused by the wedging of the bonding medium in conventional cone-type anchors. A theoretical model for the stress distribution of the variable angle multi-stage conical anchor was developed using elastic deformation theory. The reliability of this model was validated through finite element analysis. Subsequently, a comparative analysis was conducted to evaluate the stress distribution and anchoring performance of the variable angle multi-stage conical anchor against several traditional anchor types. This analysis revealed the mechanical mechanisms and advantages of the variable angle multi-stage conical anchor in enhancing anchoring effectiveness. The study illustrates that the bonding medium between adjacent cone sections of the variable angle multi-stage conical anchor exhibits a specific gap under stress, leading to reduced stress in the tendon at the connection points. This ensures a smooth transition that prevents abrupt changes in stress due to sudden shape alterations, thereby guaranteeing uniform stress distribution within the anchor section. By incorporating a multi-stage cone structure and increasing the cone angle at the free end, compressive stress can be effectively transferred to the free end, mitigating the notch effect and enhancing anchoring performance. Compared to typical conical anchors, the maximum compressive stress of the variable angle multi-stage conical anchor is reduced by 53.5 %, the outer diameter of the anchor cylinder is decreased by 22.7 %, and the anchoring efficiency is improved by 12.2 %. These improvements result in a robust anchoring effect and significant size advantage. The anchor is suitable for multibar anchoring, providing a reliable solution for large tonnage and long-span cable-stayed bridges.

Influence of horizontal alignment elements on operating speed of vehicles on two-lane highways

Earlier studies acknowledged that the design speed does not principally assure consistency in the design of highway geometric elements. Methodologies for estimating operating speeds are limited to specific or local traffic conditions and design standards, as in various studies conducted on two-lane and multi-lane highways. The present study develops operating speed models of the vehicle types observed on horizontal curves and tangent sections on two-lane highways in southern parts of India. Geometric features of the curve section and adjacent tangent sections, such as the radius of the curve, curve length, deflection angle, degree of curvature, gradient, carriageway width, and shoulder width, are obtained from the field. The speed data collected at 24 locations on the highway, including curved and tangent gradient sections, are used to analyze and model operating speed. The results indicate that the inverse of the square root of the radius appears to be an influencing parameter for modeling the operating speed of vehicles. The operating speed is sensitive for a curve radius of up to 600 m; thereafter, the operating speed appears insignificant. On tangent sections, the gradient has a larger influence on the operating speeds of all vehicle types. The operating speed models are validated and compared with models available in the literature. The proposed models developed in the present study are helpful to traffic engineers for predicting the operating speed on two-lane highways.

Hybrid deep learning approach for rock tunnel deformation prediction based on spatio-temporal patterns

The ability to predict tunnel deformation holds great significance for ensuring the reliability, safety, and sustainability of tunnel structures. However, existing deformation prediction models often simplify or overlook the impact of spatial characteristics on deformation by treating it as a time series prediction issue. This study utilizes monitoring data from the Grand Canyon Tunnel and introduces an effective data-driven method for predicting tunnel deformation based on the spatio-temporal characteristics of the historical deformation of adjacent sections. The proposed model, a combination of graph attention network (GAT) and bidirectional long and short-term memory network (Bi-LSTM), is equipped with robust spatio-temporal predictive capabilities. Additionally, the study explores other possible spatial connections and the scalability of the model. The results indicate that the proposed model outperforms other deep learning models, achieving favorable root mean square error (RMSE), mean absolute error (MAE), and coefficient of determination (R2) values of 0.34 mm, 0.23 mm, and 0.94, respectively. The graph structure based on intuitive spatial connections proves more suitable for meeting the challenges of predicting deformation. Integrating GAT-LSTM with transfer learning technology, remains stable performance when extended to other tunnels with limited data.

An alignment algorithm using coherent twin boundaries as internal reference in 3D-EBSD.

A three-dimensional (3D) microstructural volume is reconstructed from a stack of two-dimensional sections which was obtained by serial sectioning coupled with electron back scattering diffraction (EBSD) mapping of a 316L austenitic stainless steel. A new alignment algorithm named linear translation by minimising the indicator (LTMI) is proposed to reduce the translational misalignments between adjacent sections by referencing to coherent twin boundaries which are flat and lying on {111} planes. The angular difference between the measured orientation of a flat twin boundary and that of the {111} plane is used as an indicator of the accuracy of the alignment operations. This indicator is minimised through linear translations of the centroids of triangular facets, which constitute grain boundaries at a distance not restricted by the in-plane step size of the EBSD maps. And hence the systematic trend in the translational misalignments can be effectively reduced. The LTMI alignment procedure proposed herein effectively corrects the misalignments remained by other methods on a 3D-EBSD data prepared using serial sectioning methods. The accuracy in distinguishing between coherent and incoherent twin boundaries is significantly improved.

Dataset of Registered Hematoxylin–Eosin and Ki67 Histopathological Image Pairs Complemented by a Registration Algorithm

In this work, we describe a dataset suitable for analyzing the extent to which hematoxylin–eosin (HE)-stained tissue contains information about the expression of Ki67 in immunohistochemistry staining. The dataset provides images of corresponding pairs of HE and Ki67 stainings and is complemented by algorithms for computing the Ki67 index. We introduce a dataset of high-resolution histological images of testicular seminoma tissue. The dataset comprises digitized histology slides from 77 conventional testicular seminoma patients, obtained via surgical resection. For each patient, two physically adjacent tissue sections are stained: one with hematoxylin and eosin, and one with Ki67 immunohistochemistry staining. This results in a total of 154 high-resolution images. The images are provided in PNG format, facilitating ease of use for image analysis compared to the original scanner output formats. Each image contains enough tissue to generate thousands of non-overlapping 224 × 224 pixel patches. This shows the potential to generate more than 50,000 pairs of patches, one with HE staining and a corresponding Ki67 patch that depicts a very similar part of the tissue. Finally, we present the results of applying a ResNet neural network for the classification of HE patches into categories according to their Ki67 label.

Deregulation of MMP-2 and MMP-9 in laryngeal cancer: A retrospective observational study.

Laryngeal carcinoma (LC) is reported to have a higher incidence rate among all types of head and neck cancers around the globe. Mechanisms resulting in the pathogenesis of LC are complicated due to involvement of invasion and metastasis and there is a need to understand this complicated multistep process. Numerous molecules including matrix metalloproteinases (MMPs) are involved in regulating metastatic mechanisms. Furthermore, activation and expression of different classes of MMPs have been observed in multiple pathological and physiological events including inflammation, invasion, and metastasis. Among all members of MMPs, matrix metalloproteinases-2 (MMP-2), and matrix metalloproteinases-9 (MMP-9) have been frequently reported to correlate with tumor pathogenesis. The present study is designed to check the involvement of MMP-2 and MMP-9 in LC pathogenesis. 184 laryngeal tumor samples along with adjacent uninvolved healthy sections were collected to check the expression deregulation of the above-mentioned gene in LC using real-time PCR and immunohistochemistry (IHC). Real-time PCR and IHC analyses showed the significant upregulation of MMP-2 (P < .0001) and MMP-9 (P < .0001) genes in laryngeal tumors compared to controls. Spearman correlation showed the positive correlation of expression deregulation of selected MMPs with advanced TNM stage [MMP-2, (P < .0001); MMP-9, P < .0001] and smoking status [MMP-2 (P < .0001); MMP-9 P < .0001] in laryngeal pathogenesis. Receiver operating curve (ROC) analysis showed the good diagnostic/prognostic value of said markers in laryngeal cancer patients. The present study showed that significant upregulation of selected MMPs was found associated with an increased risk of laryngeal cancer and can act as good diagnostic markers for the detection of said disease.

Label-free investigation of infected acute pyelonephritis tissue by FTIR microspectroscopy with unsupervised and supervised analytical methods

Acute pyelonephritis (AP) is a severe urinary tract infection (UTI) syndrome with a large population of patients worldwide. Current approaches to confirming AP are limited to urinalysis, radiological imaging methods and histological assessment. Fourier transform infrared (FTIR) microspectroscopy is a promising label-free modality that can offer information about both morphological and molecular pathologic alterations from biological tissues. Here, FTIR microspectroscopy serves to investigate renal biological histology of a rat model with AP and classify normal cortex, normal medulla and infected acute pyelonephritis tissues. The spectra were experimentally collected by FTIR with an infrared Globar source through raster scanning procedure. Unsupervised analysis methods, including integrating, clustering and principal component analysis (PCA) were performed on such spectra data to form infrared histological maps of entire kidney section. In comparison to Hematoxylin & Eosin-stained results of the adjacent tissue sections, these infrared maps were proved to enable the differentiation of the renal tissue types. The results of both integration and clustering indicated that the concentration of amide II decreases in the infected acute pyelonephritis tissues, with an increased presence of nucleic acids and lipids. By means of PCA, the infected tissue was linearly separated from normal ones by plotting confident ellipses with the score values of the first and second principal components. Moreover, supervised analysis was performed based on the supported vector machines (SVM). Normal cortex, normal medulla and infected acute pyelonephritis tissues were classified by SVM models with the best accuracy of 96.11% in testing dataset. In addition, these analytical methods were further employed on synchrotron-based FTIR spectra data and successfully form high-resolution infrared histological maps of glomerulus and necrotic cell mass. This work demonstrates that FTIR microspectroscopy will be a powerful manner to investigate AP tissue and differentiate infected tissue from normal tissue in a renal infected model system.

Geophysical Surveys to Highlight Buried Ancient Walls of Ugento (Lecce, Italy)

Geophysics is a fundamental tool to detect buried structures of archaeological interest through non-destructive techniques. The Messapian city walls in Ugento (Puglia, southern Italy) are of great archaeological importance, and some sections are still visible. In order to locate a stretch of the city walls, geophysical prospections were performed using the low-frequency electromagnetic method and ground-penetrating radar. The surveys were carried out in a peripheral area of Ugento, near a visible section of the city walls. The analysis and interpretation of the measured data revealed clear anomalies that could be ascribed to the city walls that aligned with an adjacent section of the visible walls. Archaeological excavation campaigns found a part of the walls and some important elements, as identified by the geophysical data interpretation.

Local buckling of the bottom flange in steel-concrete composite beams with trapezoidal corrugated webs subjected to end restraint at high temperatures

Steel-concrete composite beams with trapezoidal corrugated webs (SCBCW) are prone to rapid failure at high temperatures due to local instability of the bottom flange. The failure mechanism of SCBCW, which is an integral part of the holistic structure and subject to axial and rotational restraints from adjacent members at elevated temperatures, differs from conventionally simply supported steel-concrete composite beams with straight webs. To investigate the evolution of local buckling in the compressed flange of SCBCW under high temperature conditions, a composite beam model with frame restraints was developed using ABAQUS finite element software. In this model, the composite beam is connected to restraint frame columns through high-strength bolts. The changes in axial stress, bending moment, and deflection of the compressed flange of SCBCW were analyzed as temperature increased. Results indicate that at elevated temperatures, additional bending moments occur in the flange plane due to axial restraints, section temperature gradients, section negative bending moments and accordion effect caused by corrugated web on compressed bottom flange of SCBCW. This leads to a gradual increase in axial compressive stress before local buckling occurs followed by a decrease due to internal force redistribution upon onset of buckling. Studies show that initial buckling of bottom flange occurs on one side of flat section of web where axial compressive stress is relatively high while restraint effect on web is low; Subsequent local buckling of the beam then appears on opposite side adjacent flat section of web after initial buckle occure on one side. After local buckling appears on both sides of web tension bands appear on both sides where bottom flange experiences localized bucking. The setting of longitudinal stiffeners (LS) can effectively improve the local stability of the bottom flange of the composite beam, while the setting of transverse stiffeners (TS) has no significant impact.

An open-source analysis workflow for geometrically imperfect bolted ring-flanges in wind turbine support structures

Bolted ring-flanges connect structures via two circular flanges, secured with pre-loaded bolts. In offshore wind turbine support structures the flanges connect the monopile and transition piece, the transition piece and tower, and adjacent tower sections. Fatigue analysis of the bolted ring-flanges in offshore wind turbine support structures is a critical step during design. The consideration of geometric imperfections of the flanges can be an influential factor in assessed fatigue damage over some given Fatigue Limit State load case. Schemes to analyse the effect of geometric imperfections using Finite Element Method simulations are available in literature. These schemes rely on the use of proprietary tools for the analysis. In this paper we introduce an open-source workflow to assess load transfer through geometrically imperfect bolted ring-flanges. The workflow relies on four open-source toolsets: the FreeCAD 3D parametric modelling package, the Code Aster / Salome-Meca Finite Element Method simulation environment, the ParaView post-processing and visualisation package, and the Python programming language. We present a case study of load transfer assessment in a monopile to transition piece bolted ring-flange typical of modern offshore wind turbine support structures. We decompose the load transfer across the flange under a number of geometric imperfection scenarios to determine the expected stress ranges for fatigue analysis. The results are compared with analytical methods specified in engineering standards. The workflow is configurable as per user needs, and is maintained as a GitHub repository at https://github.com/jhjorg/OpenBoltRF.

A Scalable Histological Method to Embed and Section Multiple Brains Simultaneously.

The preparation and processing of rodent brains for evaluation by immunohistochemistry is time-consuming. A large number of mouse brains are routinely used in experiments in neuroscience laboratories to evaluate several models of human diseases. Thus, methods are needed to reduce the time associated with processing brains for histology. A scalable method was developed to embed, section, and stain multiple mouse brains using supplies found in any common histology laboratory. Section collection schemes can be scaled to provide identical bregma locations between adjacent sections for immunohistochemistry, facilitating comprehensive, high-quality immunohistochemistry. As a result, sectioning and staining times are considerably reduced as sections from multiple blocks are stained simultaneously. This method improves on previous procedures and allows multiple embedding and subsequent immunostaining of brains easily with a dramatically reduced time requirement. Furthermore, we expand this method for use in numerous mouse tissues, rat brain tissue, and post-mortem human brain and arterial tissues. In summary, this procedure allows the processing of many rodent or human tissues from perfusion through microscopy in 10 days or less.

Co-localization of the sodium-glucose co-transporter-2 channel (SGLT-2) with endothelin ETA and ETB receptors in human cardiorenal tissue.

Immunocytochemistry localised SGLT-2, ETA and ETB receptors in sections of histologically normal kidney, left ventricle from patients undergoing heart transplantation or controls. Primary antisera were visualised using fluorescent microscopy. Image analysis was used to measure intensity compared with background in adjacent control sections. As expected, SGLT-2 localised to epithelial cells of the proximal convoluted tubules, and co-localised with both ET receptor sub-types. Similarly, ETA receptors predominated in cardiomyocytes; low (compared with kidney but above background) positive staining was also detected for SGLT-2. Whether low levels of SGLT-2 have a (patho)physiological role in cardiomyocytes is not known but results suggest the effect of direct blockade of sodium (and glucose) influx via SGLT-2 inhibition in cardiomyocytes should be explored, with potential for additive effects with ETA antagonists.