The adhesion GPCR GPR116/ADGRF5 has a dual function in pancreatic islets regulating somatostatin release and islet development.

Glucose homeostasis is maintained by hormones secreted from different cell types of the pancreatic islets and controlled by manifold input including signals mediated through G protein-coupled receptors (GPCRs). RNA-seq analyses revealed expression of numerous GPCRs in mouse and human pancreatic islets, among them Gpr116/Adgrf5. GPR116 is an adhesion GPCR mainly found in lung and required for surfactant secretion. Here, we demonstrate that GPR116 is involved in the somatostatin release from pancreatic delta cells using a whole-body as well as a cell-specific knock-out mouse model. Interestingly, the whole-body GPR116 deficiency causes further changes such as decreased beta-cell mass, lower number of small islets, and reduced pancreatic insulin content. Glucose homeostasis in global GPR116-deficient mice is maintained by counter-acting mechanisms modulating insulin degradation. Our data highlight an important function of GPR116 in controlling glucose homeostasis.

[Ultrasonographic findings of the stomach in warmblood horses at different filling states]. / Die sonografische Darstellung des Magens beim Warmblutpferd bei unterschiedlichen Füllungszuständen.

SUBJECT AND AIM: The aim of the study was to clarify to what extent the sonographic representation of the stomach changes depending on its filling state. MATERIAL AND METHODS: In a prospective controlled study, warmblood horses presented for gastroscopy were assessed sonographically. The examinations took place when the horses were fed normally (measurement 1), after 12 hours of food deprivation (measurement 2), after insufflation of air during gastroscopy (measurement 3) and after removal of air from the stomach at the end of the gastroscopy (measurement 4). The following objective parameters were recorded: Number of intercostal spaces (craniocaudal extension) and determination in which intercostal spaces the stomach could be visualized sonographically, maximum dorsal extension of the stomach, distance between skin and stomach wall. RESULTS: The median number of intercostal spaces in which the stomach could be visualized sonographically was 7.5 (IQR 3.75), 1.0 (IQR 1.0), 7.0 (IQR 2.0) and 2.0 (IQR 1.0) for measurements 1, 2, 3 (n=32) and 4 (n=15), respectively. The differences were significant between measurements 1 and 2 and between measurements 2 and 3. There was no significant difference between measurements 1 and 3. After 12 hours of food deprivation, the stomach size measured by the number of intercostal spaces visualizing the stomach was reduced by 75%. The maximum dorsal expansion of the stomach for measurements 1, 2, 3 (n=32) and 4 (n=15) was a median of 38 cm (IQR 15.25), 13 cm (IQR 6.75), 43 cm (IQR 7.00) and 21 cm (IQR 8.00), respectively. The differences were significant between measurements 1 and 2 and between 2 and 3. Concerning the distance between skin and stomach wall, the following medians were determined for measurements 1, 2, 3 (n=32) and 4 (n=15): 5.8 cm (IQR 2.27), 4.05 cm (IQR 3.05), 4.8 cm (IQR 1.48) and 5.9 cm (IQR 2.90). The only statistically significant difference was observed between measurements 1 and 3. CONCLUSIONS AND CLINICAL RELEVANCE: The sonographic appearance of the stomach changes according to the state of filling. Parameters that are readily determined sonographically are the craniocaudal and maximum dorsal extension as well as the distance between the skin and the stomach wall.

Accuracy Analysis of Computer-Assisted and Guided Dental Implantology by Comparing 3D Planning Data and Actual Implant Placement in a Mandibular Training Model: A Monocentric Comparison between Dental Students and Trained Implantologists.

The aim of this study was to investigate how precisely implantation can be realized by participants on a phantom head according to preliminary planning. Of particular interest here was the influence of participants' previous knowledge and surgical experience on the precision of the implant placement. The placed implants were scanned using an intraoral scanner, saved as STL files, and superimposed with the 3D-planned implant placement. Deviations from the planning were indicated in millimeters and degrees. We were able to show that on average, the deviations from computer-assisted 3D planning were less than 1 mm for implantologists, and the students also did not deviate more than 1.78 mm on average from 3D planning. This study shows that guided implantology provides predictable and reproducible results in dental implantology. Incorrect positioning, injuries to anatomical structures, and implant positions that cannot be prosthetically restored can thus be avoided.

Taxane-Induced Neuropathy and Its Ocular Effects-A Longitudinal Follow-up Study in Breast Cancer Patients.

A common severe neurotoxic side effect of breast cancer (BC) therapy is chemotherapy-induced peripheral neuropathy (CIPN) and intervention is highly needed for the detection, prevention, and treatment of CIPN at an early stage. As the eye is susceptible to neurotoxic stimuli, the present study aims to determine whether CIPN signs in paclitaxel-treated BC patients correlate with ocular changes by applying advanced non-invasive biophotonic in vivo imaging. Patients (n = 14, 10 controls) underwent monitoring sessions after diagnosis, during, and after therapy (T0-T3). Monitoring sessions included general anamnesis, assessment of their quality of life, neurological scores, ophthalmological status, macular optical coherence tomography (OCT), and imaging of their subbasal nerve plexus (SNP) by large-area confocal laser-scanning microscopy (CLSM). At T0, no significant differences were detected between patients and controls. During treatment, patients' scores significantly changed while the greatest differences were found between T0 and T3. None of the patients developed severe CIPN but retinal thickenings could be detected. CLSM revealed large SNP mosaics with identical areas while corneal nerves remained stable. The study represents the first longitudinal study combining oncological examinations with advanced biophotonic imaging techniques, demonstrating a powerful tool for the objective assessment of the severity of neurotoxic events with ocular structures acting as potential biomarkers.

Dental Root Injuries Caused by Osteosynthesis Screws in Orthognathic Surgery-Comparison of Conventional Osteosynthesis and Osteosynthesis by CAD/CAM Drill Guides and Patient-Specific Implants.

BACKGROUND/AIM: The primary aim was to evaluate the prevalence and localisation of dental injuries caused by osteosynthesis screws during orthognathic surgery, comparing two different CAD/CAM planning/surgical approaches through retrospective evaluation of post-operative computed tomography. MATERIAL AND METHODS: This study considered all patients who underwent orthognathic surgery from 2010-2019. The examination for dental root injuries between conventional osteosynthesis (Maxilla conventional cohort) and osteosynthesis with patient-specific implant (Maxilla PSI cohort) was performed by evaluating the post-operative CT scans. RESULTS: A total of 126 patients were included in the study. Among the 61 patients of the Maxilla conventional cohort, 10 dental root injuries in 8 patients (13.1%) were detected in the post-operative CT scan, representing 1.5% (n = 10/651) of the osteosynthesis screws inserted in proximity of the alveolar crest. No dental injury occurred following osteosynthesis in the 65 patients of the Maxillary PSI cohort (n = 0/773 screws) (p < 0.001). During a mean follow-up period of 13 months after primary surgery, none of the injured teeth showed evidence of periapical alterations and no endodontic treatments were necessary. CONCLUSIONS: Maxillary positioning using CAD/CAM-fabricated drill/osteotomy guide and osteosynthesis with PSI can significantly reduce the risk for dental injury compared to the conventional procedure. However, the clinical significance of the detected dental injuries was rather minor.

Reduced urine volume and changed renal sphingolipid metabolism in P2ry14-deficient mice.

The UDP-glucose receptor P2RY14, a rhodopsin-like G protein-coupled receptor (GPCR), was previously described as receptor expressed in A-intercalated cells of the mouse kidney. Additionally, we found P2RY14 is abundantly expressed in mouse renal collecting duct principal cells of the papilla and epithelial cells lining the renal papilla. To better understand its physiological function in kidney, we took advantage of a P2ry14 reporter and gene-deficient (KO) mouse strain. Morphometric studies showed that the receptor function contributes to kidney morphology. KO mice had a broader cortex relative to the total kidney area than wild-type (WT) mice. In contrast, the area of the outer stripe of the outer medulla was larger in WT compared to KO mice. Transcriptome comparison of the papilla region of WT and KO mice revealed differences in the gene expression of extracellular matrix proteins (e.g., decorin, fibulin-1, fibulin-7) and proteins involved in sphingolipid metabolism (e.g., small subunit b of the serine palmitoyltransferase) and other related GPCRs (e.g., GPR171). Using mass spectrometry, changes in the sphingolipid composition (e.g., chain length) were detected in the renal papilla of KO mice. At the functional level, we found that KO mice had a reduced urine volume but an unchanged glomerular filtration rate under normal chow and salt diets. Our study revealed P2ry14 as a functionally important GPCR in collecting duct principal cells and cells lining the renal papilla and the possible involvement of P2ry14 in nephroprotection by regulation of decorin.

Comparison of Anatomical Preformed Titanium Implants and Patient-Specific CAD/CAM Implants in the Primary Reconstruction of Isolated Orbital Fractures-A Retrospective Study.

BACKGROUND/AIM: Reconstruction of the fractured orbit remains a challenge. The aim of this study was to compare anatomical preformed titanium orbital implants with patient-specific CAD/CAM implants for precision and intraoperative applicability. MATERIAL AND METHODS: A total of 75 orbital reconstructions from 2012 to 2022 were retrospectively assessed for their precision of implant position and intra- and postoperative revision rates. For this purpose, the implant position after digital orbital reconstruction was checked for deviations by mirroring the healthy orbit at 5 defined points, and the medical records of the patients were checked for revisions. RESULTS: The evaluation of the 45 anatomical preformed orbital implant cases showed significantly higher deviations and an implant inaccuracy of 66.6% than the 30 CAD/CAM cases with only 10% inaccuracy. In particular, the CAD/CAM implants were significantly more precise in medial and posterior positioning. In addition, the intraoperative revision rates of 26.6% vs. 11% after 3D intraoperative imaging and the postoperative revision rates of 13% vs. 0 for the anatomical preformed implants were significantly higher than for patient-specific implants. CONCLUSION: We conclude that patient-specific CAD/CAM orbital implants are highly suitable for primary orbital reconstruction. These seem to be preferable to anatomical preformed implants in terms of precision and revision rates.

Approaches to standardising the magnetic resonance image analysis of equine tendon lesions.

Background: Low-field magnetic resonance imaging (MRI) has gained increasing importance to monitor equine tendon lesions. Comparing results between studies and cases is hampered, because image analysis approaches vary strongly. This study aimed to improve reliability, comparability and time efficiency of quantitative MRI image analysis. Methods: Induced tendon lesions were studied over a 24-week period with 10 follow-up MRI examinations. Signal intensities (SIs) of tendons, tendon lesions, cortical bone and background, as well as lesion cross-sectional areas (CSAs) were measured. Lesion SI standardisation with different formulas was evaluated, using histological findings as reference. Different types of region of interest (ROI) for lesion SI measurement were compared. Lesion CSA measurement at different levels was evaluated, using the calculated total lesion volume as reference. Subjective lesion identification and manual CSA and SI measurements were compared to an automated, algorithm-based approach. Results: Lesion SI standardised using a quotient of lesion and background or cortical bone SI, correlated best with histologically determined lesion severity. Lesion SI in circular ROIs correlated strongly with lesion SI in free-hand whole-lesion ROIs. The level of the maximum lesion CSA shifted over time; the CSA maximum correlated strongly with lesion volume. In sequences with short acquisition time, algorithm-based automated lesion detection showed almost perfect agreement with subjective lesion identification. Automated measurement of CSA and SI was also feasible, with stronger correlation and better agreement with the manually obtained data for the SI than for the CSA. Conclusion: Our study may provide guidance for MRI image analysis of tendon healing. Reliable image analysis can be performed time-efficiently, particularly regarding lesion SI quantification.

Mosaic vs. Single Image Analysis with Confocal Microscopy of the Corneal Nerve Plexus for Diagnosis of Early Diabetic Peripheral Neuropathy.

INTRODUCTION: The assessment of the corneal nerve fibre plexus with corneal confocal microscopy (CCM) is an upcoming but still experimental method in the diagnosis of early stage diabetic peripheral neuropathy (DPN). Using an innovative imaging technique-Heidelberg Retina Tomograph equipped with the Rostock Cornea Module (HRT-RCM) and EyeGuidance module (EG)-we were able to look at greater areas of subbasal nerve plexus (SNP) in order to increase the diagnostic accuracy. The aim of our study was to evaluate the usefulness of EG instead of single image analysis in diagnosis of early stage DPN. METHODS: This prospective study was performed on 60 patients with type 2 diabetes mellitus, classified equally into two subgroups based on neuropathy deficient score (NDS): patients without DPN (group 1) or with mild DPN (group 2). The following parameters were analysed in the two subgroups: corneal nerve fibre length (CNFL; mm/mm2), corneal nerve fibre density (CNFD; no./mm2), corneal nerve branch density (CNBD; no./mm2). Furthermore, we compared the data calculated with the novel mosaic, EG-based method with those received from single image analysis using different quantification tools. RESULTS: Using EG we did not find a significant difference between group 1 and group 2: CNFL (16.81 ± 5.87 mm/mm2 vs. 17.19 ± 7.19 mm/mm2, p = 0.895), CNFD (254.05 ± 115.36 no./mm2 vs. 265.91 ± 161.63 no./mm2, p = 0.732) and CNBD (102.68 ± 62.28 no./mm2 vs. 115.38 ± 96.91 no./mm2, p = 0.541). No significant difference between the EG method of analysing the SNP and the single image analysis of 10 images per patient was detected. CONCLUSION: On the basis of our results it was not possible to differentiate between early stages of large nerve fibre DPN in patients with type 2 diabetes mellitus via SNP analysis. To improve sensitivity and specificity of this method newer technologies are under current evaluation. TRIAL REGISTRATION: ClinicalTrials.gov Identifier NCT05326958.

Intestinal S100/Calgranulin Expression in Cats with Chronic Inflammatory Enteropathy and Intestinal Lymphoma.

Diagnosing chronic inflammatory enteropathies (CIE) in cats and differentiation from intestinal lymphoma (IL) using currently available diagnostics is challenging. Intestinally expressed S100/calgranulins, measured in fecal samples, appear to be useful non-invasive biomarkers for canine CIE but have not been evaluated in cats. We hypothesized S100/calgranulins to play a role in the pathogenesis of feline chronic enteropathies (FCE) and to correlate with clinical and/or histologic disease severity. This retrospective case-control study included patient data and gastrointestinal (GI) tissues from 16 cats with CIE, 8 cats with IL, and 16 controls with no clinical signs of GI disease. GI tissue biopsies were immunohistochemically stained using polyclonal α-S100A8/A9 and α-S100A12 antibodies. S100A8/A9+ and S100A12+ cells were detected in all GI segments, with few significant differences between CIE, IL, and controls and no difference between diseased groups. Segmental inflammatory lesions were moderately to strongly correlated with increased S100/calgranulin-positive cell counts. Clinical disease severity correlated with S100A12+ cell counts in cats with IL (ρ = 0.69, p = 0.042) and more severe diarrhea with colonic lamina propria S100A12+ cells with CIE (ρ = 0.78, p = 0.021) and duodenal S100A8/A9+ cells with IL (ρ = 0.71, p = 0.032). These findings suggest a role of the S100/calgranulins in the pathogenesis of the spectrum of FCE, including CIE and IL.

Equine skeletal scintigraphy: Comparing normal bone-to-soft tissue ratio 2 and 4 hours after injection with either hydroxymethylene diphosphonate or methylene diphosphonate.

BACKGROUND: In equine skeletal scintigraphy, there is no information about the possible influence of different phosphonate compounds on image quality. METHODS: This prospective randomised study determined bone uptake changes and image quality for hydroxymethylene diphosphonate (HDP) and methylene diphosphonate (MDP) in equine patients at different time points. Scintigraphic images of the radius and the tibia of 20 horses were acquired at 2 and 4 hours after injection of either technetium-labelled HDP or MDP. Three regions of interest were identified-in the bone diaphysis, adjacent soft tissue and background area-to determine the normal bone-to-soft tissue ratio (BSR). Qualitative analysis was performed using a modified scoring system. RESULTS: In terms of BSR and count rates, HDP showed a slightly better incorporation into bone compared to MDP, but significant differences were only detected for count rates at 4 hours after injection (p = 0.048). The radiopharmaceutical used did not influence qualitative image scoring, which was correlated with the BSR (ρ = 0.49; p ≤ 0.001). CONCLUSION: The choice of HDP or MDP for equine skeletal scintigraphy does not seem to substantially affect BSR and qualitative image scoring. Further studies with a larger sample size, including the evaluation of lesion detection ability for both bone-seeking agents, are needed.

[Venous catheters in horses - early ultrasonographic venous abnormalities and evidence of biofilms]. / Ultrasonografische Früherkennung von Venenveränderungen sowie der Nachweis von Biofilmen auf Venenverweilkathetern beim Pferd.

OBJECTIVE: Complications associated with indwelling intravenous catheters vary from minor to severe. Changes in tissue architecture and vein structure may be detectable prior to clinical alterations. The aim of the study was to characterize and compare changes in a catheterized vein and surrounding tissue by clinical and ultrasonographic examination. Microbiological infestation of catheter specimens was assessed. MATERIAL AND METHODS: In this prospective, observational, clinical study 55 horses with an indwelling intravenous catheter have been included. Subsequent to catheter placement, vein and surrounding tissue were daily examined clinically and by ultrasonographic examination at predetermined localizations. After sterile removal of the catheters, specimens underwent microbiological testing and scanning electron microscopy. Obtained data were analyzed via descriptive statistics. Thickness of the venous wall was compared at predetermined localizations and time points for several parameters with the help of non-parametric tests (level of significance at p < 0.05, post-hoc Bonferroni correction). RESULTS: Overall, in 41.8 % of the horses transient alterations (swelling) of the surrounding tissue occurred during catheterization. Median catheterization was 69.5 hours (19 hour-10 days). With ultrasonography, venous valves and collateral vessels could be detected reliably. Significant alterations in wall thickness during catheterization were associated with various factors (location site [p ≤ 0.001], season [p = 0.006], anesthesia [p ≤ 0.001]).Microbiological analysis revealed a positive result in 23.5 %, 12 of the 51 samples; raster electron microscopy showed presence of bacteria in 25.0 %, 4 of the 16 investigated catheter samples. CONCLUSION AND CLINICAL RELEVANCE: Local tissue changes at the insertion site of the catheter are commonly associated with catheterization, and are easily detectable with ultrasonography. Despite the easy performance in a clinical setting, the benefit of routine ultrasonographic monitoring of catheterized veins might be questionable with regard to early identification and prediction of catheter-associated venous disease. Bacteria might be detectable morphologically on the catheter but frequently lack a positive result from standard bacterial cultivation.

Surfactant Protein-G in Wildtype and 3xTg-AD Mice: Localization in the Forebrain, Age-Dependent Hippocampal Dot-like Deposits and Brain Content.

The classic surfactant proteins (SPs) A, B, C, and D were discovered in the lungs, where they contribute to host defense and regulate the alveolar surface tension during breathing. Their additional importance for brain physiology was discovered decades later. SP-G, a novel amphiphilic SP, was then identified in the lungs and is mostly linked to inflammation. In the brain, it is also present and significantly elevated after hemorrhage in premature infants and in distinct conditions affecting the cerebrospinal fluid circulation of adults. However, current knowledge on SP-G-expression is limited to ependymal cells and some neurons in the subventricular and superficial cortex. Therefore, we primarily focused on the distribution of SP-G-immunoreactivity (ir) and its spatial relationships with components of the neurovascular unit in murine forebrains. Triple fluorescence labeling elucidated SP-G-co-expressing neurons in the habenula, infundibulum, and hypothalamus. Exploring whether SP-G might play a role in Alzheimer's disease (AD), 3xTg-AD mice were investigated and displayed age-dependent hippocampal deposits of ß-amyloid and hyperphosphorylated tau separately from clustered, SP-G-containing dots with additional Reelin-ir-which was used as established marker for disease progression in this specific context. Semi-quantification of those dots, together with immunoassay-based quantification of intra- and extracellular SP-G, revealed a significant elevation in old 3xTg mice when compared to age-matched wildtype animals. This suggests a role of SP-G for the pathophysiology of AD, but a confirmation with human samples is required.

Histological differences between lumbar and tail intervertebral discs in mice.

Both the lumbar and tail intervertebral discs (IVD) of mice serve as models for the pathogenesis and histologic progression of degenerative disc disease. Recent studies in mature mice, however, demonstrate that the mechanics and physical attributes of lumbar and tail IVD-endplate (EP)-interfaces are strikingly different. We hypothesized that these structural disparities are associated with differences in the composition and organization of soft tissue elements that influence the biomechanical properties of the spine. Lumbar and tail vertebral segments and discs were collected from the same C57BL/6N and C57BL/6JRj mice, respectively for histological comparison of coronal sections at the ages of 4 weeks (weaned, both strains, C57BL/6N: n = 7; C57BL/6JRj: n = 4), three (mature, C57BL/6N: n = 7; C57BL/6JRj: n = 4), twelve (middle aged, C57BL/6JRj only: n = 3) and eighteen (old, C57BL/6JRj only: n = 3) months old. The histology of lumbar and tail IVD-EP-interfaces of mature mice differed markedly. The lumbar IVD-EP-interphase was characterized by a broad cartilaginous EP, while the tail IVD-EP-interphase comprised a thin layer of cartilage cells adjacent to a broad bony layer abutting the vertebral growth plate. Furthermore, the composition of the nuclei pulposi (NP) of lumbar and tail IVD in mature mice differed greatly. Lumbar NP consisted of a compact cluster of mainly large, uni-vacuolated cells centered in an amorphous matrix, while tail NP were composed of a loose aggregate of vacuolated and non-vacuolated cells. The anuli fibrosi also differed, with more abundant and sharply defined lamellae in tail compared to lumbar discs. The observed histological differences in the EP were even most prominent in weaned mice but were still discernible in middle-aged and old mice. An appreciation of the histological differences between lumbar and tail IVD components in mice, including nucleus pulposus, annulus fibrosus, and endplates, is essential to our understanding of spinal biomechanics in these animals and should inform the design and interpretation of future IVD-studies.

3T MRI characteristics of the palatine tonsil in brachycephalic dogs.

BACKGROUND: Investigation of the MRI characteristics of the palatine tonsil in brachycephalic dogs in 3T high-field system. METHODS: Eighty-five brachycephalic dogs and 37 normocephalic dogs were divided into five groups: group 1 French bulldogs (FBs) with neurological clinical signs (n = 37), group 2 FBs with brachycephalic obstructive airway syndrome (BOAS) (n = 22), group 3 pugs with neurological clinical signs (n = 17), group 4 pugs with BOAS (n = 9) and group 5 normocephalic dogs (n = 37). Cross-sectional area and volume measurements were performed, and tonsillar margination and contour, shape, signal intensity and homogeneity/heterogeneity of the palatine tonsils were evaluated and compared. RESULTS: Cross-sectional area and volume measurements of the tonsils showed no significant differences between brachycephalic and normocephalic dogs with the exception of the dogs of group 2 (FB BOAS), which showed relatively high volume and large cross-sectional area in comparison to other groups. In 87% of the brachycephalic animals, the tonsils were well defined. A smooth contour was detectable in 91.8% and a rounded shape in 94.7% of brachycephalic dogs. Signal intensity was assessed as hyperintense in relation to the musculature and iso- to hyperintense to the soft palate. Heterogeneous appearance was described in 86.9% of the brachycephalic animals. CONCLUSIONS: The MRI characteristics of the tonsils of brachycephalic dogs do not differ considerably from those of normocephalic dogs. In FBs with distinct clinical signs of obstructive airway syndrome, increase in cross-sectional area and volume of the tonsils was detected.

RNA sequencing of glioblastoma tissue slice cultures reveals the effects of treatment at the transcriptional level.

One of the major challenges in cancer research is finding models that closely resemble tumors within patients. Human tissue slice cultures are a promising approach to provide a model of the patient's tumor biology ex vivo. Recently, it was shown that these slices can be successfully analyzed by whole transcriptome sequencing as well as automated histochemistry, increasing their usability as preclinical model. Glioblastoma multiforme (GBM) is a highly malignant brain tumor with poor prognosis and little is known about its genetic background and heterogeneity regarding therapy success. In this study, tissue from the tumors of 25 patients with primary GBM was processed into slice cultures and treated with standard therapy (irradiation and temozolomide). Total RNA sequencing and automated histochemistry were performed to enable analysis of treatment effects at a transcriptional and histological level. Slice cultures from long-term survivors (overall survival [OS] > 24 months) exhibited more apoptosis than cultures from patients with shorter OS. Proliferation within these slices was slightly increased in contrast to other groups, but not significantly. Among all samples, 58 protein-coding genes were upregulated and 32 downregulated in treated vs. untreated slice cultures. In general, an upregulation of DNA damage-related and cell cycle checkpoint genes as well as enrichment of genotoxicity pathways and p53-dependent signaling was found after treatment. Overall, the current study reproduces knowledge from former studies regarding the feasibility of transcriptomic analyses and automated histology in tissue slice cultures. We further demonstrate that the experimental data merge with the clinical follow-up of the patients, which improves the applicability of our model system.

Droplet Degeneration of Hippocampal and Cortical Neurons Signifies the Beginning of Neuritic Plaque Formation.

BACKGROUND: Neuritic plaques contain neural and microglial elements, and amyloid-ß protein (Aß), but their pathogenesis remains unknown. OBJECTIVE: Elucidate neuritic plaque pathogenesis. METHODS: Histochemical visualization of hyperphosphorylated-tau positive (p-tau+) structures, microglia, Aß, and iron. RESULTS: Disintegration of large projection neurons in human hippocampus and neocortex presents as droplet degeneration: pretangle neurons break up into spheres of numerous p-tau+ droplets of various sizes, which marks the beginning of neuritic plaques. These droplet spheres develop in the absence of colocalized Aß deposits but once formed become encased in diffuse Aß with great specificity. In contrast, neurofibrillary tangles often do not colocalize with Aß. Double-labelling for p-tau and microglia showed a lack of microglial activation or phagocytosis of p-tau+ degeneration droplets but revealed massive upregulation of ferritin in microglia suggesting presence of high levels of free iron. Perl's Prussian blue produced positive staining of microglia, droplet spheres, and Aß plaque cores supporting the suggestion that droplet degeneration of pretangle neurons in the hippocampus and cortex represents ferroptosis, which is accompanied by the release of neuronal iron extracellularly. CONCLUSION: Age-related iron accumulation and ferroptosis in the CNS likely trigger at least two endogenous mechanisms of neuroprotective iron sequestration and chelation, microglial ferritin expression and Aß deposition, respectively, both contributing to the formation of neuritic plaques. Since neurofibrillary tangles and Aß deposits colocalize infrequently, tangle formation likely does not involve release of neuronal iron extracellularly. In human brain, targeted deposition of Aß occurs specifically in response to ongoing ferroptotic droplet degeneration thereby producing neuritic plaques.

Leptin Receptor-Deficient db/db Mice Show Significant Heterogeneity in Response to High Non-heme Iron Diet.

Recent studies have shown an association between iron homeostasis, obesity and diabetes. In this work, we investigated the differences in the metabolic status and inflammation in liver, pancreas and visceral adipose tissue of leptin receptor-deficient db/db mice dependent on high iron concentration diet. 3-month-old male BKS-Leprdb/db/JOrlRj (db/db) mice were divided into two groups, which were fed with different diets containing high iron (29 g/kg, n = 57) or standard iron (0.178 g/kg; n = 42) concentrations for 4 months. As anticipated, standard iron-fed db/db mice developed obesity and diabetes. However, high iron-fed mice exhibited a wide heterogeneity. By dividing into two subgroups at the diabetes level, non-diabetic subgroup 1 (<13.5 mmol/l, n = 30) significantly differed from diabetic subgroup two (>13.5 mmol/l, n = 27). Blood glucose concentration, HbA1c value, inflammation markers interleukin six and tumor necrosis factor α and heme oxygenase one in visceral adipose tissue were reduced in subgroup one compared to subgroup two. In contrast, body weight, C-peptide, serum insulin and serum iron concentrations, pancreatic islet and signal ratio as well as cholesterol, LDL and HDL levels were enhanced in subgroup one. While these significant differences require further studies and explanation, our results might also explain the often-contradictory results of the metabolic studies with db/db mice.

Surfactant protein C is associated with perineuronal nets and shows age-dependent changes of brain content and hippocampal deposits in wildtype and 3xTg mice.

Surfactant protein C (SP-C) modulates cerebrospinal fluid (CSF) rheology. During ageing, its declining levels are accompanied by an increased burden of white matter lesions. Pulmonary SP-C intermediates harbouring the BRICHOS-domain prevent protein misfolding in the lungs. Thus, cerebral SP-C intermediates may counteract cerebral ß-amyloidosis, a hallmark of Alzheimer's disease (AD). However, data on the molecular neuroanatomy of SP-C and its alterations in wildtype and triple transgenic (3xTg) mice, featuring essential elements of AD-neuropathology, are lacking. Therefore, this study investigated SP-C-containing structures in murine forebrains and their spatial relationships with vascular, glial and neuronal components of the neurovascular unit. Fluorescence labelling demonstrated neuronal SP-C in the medial habenula, the indusium griseum and the hippocampus. Glial counterstaining elucidated astrocytes in the corpus callosum co-expressing SP-C and S100ß. Notably, perineuronal nets were associated with SP-C in the nucleus reticularis thalami, the lateral hypothalamus and the retrosplenial cortex. In the hippocampus of aged 3xTg mice, an increased number of dot-like depositions containing SP-C and Reelin, but devoid of BRICHOS-immunoreactivity were observed apart from AD-like lesions. Wildtype and 3xTg mice revealed an age-dependent increase of such deposits markedly pronounced in about 24-month-old 3xTg mice. SP-C levels of the intracellular and extracellular compartments in each group revealed an inverse correlation of SP-C and Reelin, with reduced SP-C and increased Reelin in an age-dependent fashion especially in 3xTg mice. Taken together, extracellular SP-C, as modulator of glymphatic clearance and potential ligand of PNs, declines in 3xTg mice, which show an accumulation of extracellular Reelin depositions during ageing.

Long-term diet-induced obesity does not lead to learning and memory impairment in adult mice.

Obesity arising from excessive dietary fat intake is a risk factor for cognitive decline, dementia and neurodegenerative diseases, including Alzheimer's disease. Here, we studied the effect of long-term high-fat diet (HFD) (24 weeks) and return to normal diet (ND) on behavioral features, microglia and neurons in adult male C57BL/6J mice. Consequences of HFD-induced obesity and dietary changes on general health (coat appearance, presence of vibrissae), sensory and motor reflexes, learning and memory were assessed by applying a phenotypic assessment protocol, the Y maze and Morris Water Maze test. Neurons and microglia were histologically analyzed within the mediobasal hypothalamus, hippocampus and frontal motor cortex after long-term HFD and change of diet. Long periods of HFD caused general health issues (coat alterations, loss of vibrissae), but did not affect sensory and motor reflexes, emotional state, memory and learning. Long-term HFD increased the microglial response (increased Iba1 fluorescence intensity, percentage of Iba1-stained area and Iba1 gene expression) within the hypothalamus, but not in the cortex and hippocampus. In neither of these regions, neurodegeneration or intracellular lipid droplet accumulation was observed. The former alterations were reversible in mice whose diet was changed from HFD to ND. Taken together, long periods of excessive dietary fat alone do not cause learning deficits or spatial memory impairment, though HFD-induced obesity may have detrimental consequences for cognitive flexibility. Our data confirm the selective responsiveness of hypothalamic microglia to HFD.