Deep Laminae Research Articles

CNSC-65. ELECTROPHYSIOLOGICAL AND SPATIAL TRANSCRIPTOMIC PROFILING OF GLIOMA-INFILTRATED CORTEX REVEAL LAYER SPECIFIC ALTERATIONS

Abstract Specialized neocortical circuits selectively organize neuronal signals and encode features of cognitive processing via local and long-range connections. Since glioma-infiltrated cortex is excitable and can participate in cognitive processing, the underlying laminar structure and functionality may still be preserved despite infiltration. Moreover, since glioma cells remodel existing neural circuits and microenvironmental factors drive invasion and proliferation, neuron-glioma interactions may demonstrate layer specificity. As glioma-infiltrated cortex remains poorly understood, we sought to characterize these regions using electrophysiological, structural, and genomic approaches. We assessed the power spectra of normal-appearing and glioma-infiltrated cortex using magnetoencephalography and subdural high-density electrode arrays. Immunohistochemistry of formalin-fixed paraffin-embedded (FFPE) samples of infiltrated cortex enabled protein-level neuronal and glioma identification to determine laminar preservation and spatial invasion patterns. Spatial transcriptomics of FFPE tissues and single-nucleus RNA-sequencing were used to identify cell populations, genomic alterations, and cell-cell communication within and across samples at testing and validation sites across the US and Europe. Increased delta range (1-4 Hz) power and decreased power in the beta range (12-20 Hz), activity thought to originate from deep layers, were identified as robust features of infiltrated cortex which were maintained across glioma subtypes (n = 139 patients) and preserved in a validation cohort (n = 8 patients). Immunohistochemistry revealed greater tumor burden within deep laminae regardless of glioma subtype (n = 20 samples). Tissue proteomics and spatial genomics analyses performed using glioma-infiltrated cortical samples (n = 10 samples; 29,011 spots) confirmed routine preservation of laminar organization, greater tumor burden in deep layers, as well as layer specific differences in glioma-related expression programs. Cell-cell communication analyses demonstrated increased interactions in glioma-infiltrated cortex across layers. These findings suggest that laminar structure may be preserved in glioma-infiltrated cortex while remodeling of neural circuits alters spatiotemporal activity in a predictable manner and support a deep to superficial invasion pattern. Thus, investigating glioma infiltration using layer-centric approaches may facilitate novel insights into neuron-glioma interactions.

Vertebrate paleontology of the Cloverly Formation (Lower Cretaceous), IV: the oldest edentulous frog (Salientia) from Laurasia

ABSTRACT Edentulism—lack of teeth—is a derived condition among salientians (total-group frogs and toads) that has arisen independently at least 22 times within the crown group, Anura. Despite this frequency, edentulism (in part or complete) is seldom documented in the salientian fossil record, and thus its evolutionary history remains obscure. A ∼90 Ma gap presently exists between the edentulous salientians Notobatrachus reigi (late Toarcian; Argentina) and the five known species from the latest Cretaceous of Argentina, South Africa, Canada, and the U.S.A. Here we report a new instance of edentulism in an Early Cretaceous salientian, Ostrombatrachos nodos gen. et sp. nov., based on a maxilla from an early Albian age (∼ 111 Ma) locality in the Cloverly Formation of Wyoming, U.S.A. Although incomplete, the holotype maxilla exhibits a distinctive suite of features (i.e., edentulous; pit-and-ridge external ornament; shallow groove across anteriormost portion of margo orbitalis and continuing anteroventrally down lateral surface of pars facialis to interrupt the external ornament; medially expanded orbital flange; moderately broad, deep lamina horizontalis; and weakly developed processus pterygoideus) that is unknown in any other salientian. This new taxon represents the oldest occurrence of edentulous maxillae in a Laurasian salientian and helps fill the long temporal gap in edentulism among salientians. O. nodos gen. et sp. nov. does not appear to be closely related to other edentulous Mesozoic salientians, suggesting it represents yet another independent evolution of this feature.

Mixed adenoma-neuroendocrine tumor of the stomach: analysis of nine cases with literature review.

Mixed adenoma-neuroendocrine tumor (MANET) comprises adenoma and well-differentiated neuroendocrine tumor (NET) components. Given the limited information on this due to its rarity, we aimed to clarify the clinicopathologic features and optimal management of gastric MANETs in a case series and literature review. Nine patients with gastric MANETs, including eight male and one female patient (mean age, 72years), were identified from the institutional pathology archive. Endoscopically, the tumors appeared as flat elevated lesions with sizes ranging from 0.8 to 4.4cm. One patient had familial adenomatous polyposis, and no patient had autoimmune gastritis. All MANETs developed in the gastric body mucosa exhibiting chronic metaplastic atrophic gastritis. The glandular components were intestinal-type low-grade adenoma, and focal high-grade dysplasia was also recognized in three cases. The NET component was in middle/deep lamina propria in six cases and confined to deep lamina propria in the remaining three cases. Minimal cytologic atypia was found in the NET component, with no recognizable mitosis and a Ki-67 labeling index of < 2%. The NET component mostly showed diffuse positivity for serotonin and CDX2, suggesting that it consists of enterochromaffin cells. Diffuse p53 immunostaining was observed only in the high-grade adenomatous component of one case. No recurrence was observed during the follow-up period of 2-94months. Correct distinction between the NET and poorly differentiated carcinoma components is crucial to prevent overtreatment of gastric MANETs. Considering its indolent nature, endoscopic resection is the primary recommendation for gastric MANETs as well as for pure adenomas.

Deep sequencing of Phox2a nuclei reveals five classes of anterolateral system neurons

The anterolateral system (ALS) is a major ascending pathway from the spinal cord that projects to multiple brain areas and underlies the perception of pain, itch, and skin temperature. Despite its importance, our understanding of this system has been hampered by the considerable functional and molecular diversity of its constituent cells. Here, we use fluorescence-activated cell sorting to isolate ALS neurons belonging to the Phox2a-lineage for single-nucleus RNA sequencing. We reveal five distinct clusters of ALS neurons (ALS1-5) and document their laminar distribution in the spinal cord using in situ hybridization. We identify three clusters of neurons located predominantly in laminae I-III of the dorsal horn (ALS1-3) and two clusters with cell bodies located in deeper laminae (ALS4 and ALS5). Our findings reveal the transcriptional logic that underlies ALS neuronal diversity in the adult mouse and uncover the molecular identity of two previously identified classes of projection neurons. We also show that these molecular signatures can be used to target groups of ALS neurons using retrograde viral tracing. Overall, our findings provide a valuable resource for studying somatosensory biology and targeting subclasses of ALS neurons.

Development and growth of the temporal fascia: a histological study using human fetuses.

The temporal fascia is a double lamina sandwiching a thick fat layer above the zygomatic bony arch. To characterize each lamina, their developmental processes were examined in fetuses. We observed histological sections from 22 half-heads of 10 mid-term fetuses at 14-18 weeks (crown-rump length, 95-150 mm) and 12 near-term fetuses at 26-40 weeks (crown-rump length, 215-334 mm). The superficial lamina of the temporal fascia was not evident at mid-term. Instead, a loose subcutaneous tissue was attached to the thin, deep lamina of the temporal fascia covering the temporalis muscle. At near-term, the deep lamina became thick, while the superficial lamina appeared and exhibited several variations: i) a mono-layered thick membrane (5 specimens); ii) a multi-layered membranous structure (6) and; iii) a cluster of independent thick fasciae each of which were separated by fatty tissues (1). In the second and third patterns, fatty tissue between the two laminae was likely to contain longitudinal fibrous bands in parallel with the deep lamina. Varying proportions of the multi-layered superficial lamina were not attached to the zygomatic arch, but extended below the bony arch. Whether or not lobulation or septation of fatty tissues was evident was not dependent on age. The deep lamina seemed to develop from the temporalis muscle depending on the muscle contraction. In contrast, the superficial lamina developed from subcutaneous collagenous bundles continuous to the cheek. Therein, a difference in development was clearly seen between two categories of the fasciae.

Concomitant use of pre-emptive analgesia with local and general anesthesia in rat uterine pain surgical model.

Preemptive analgesia is used for postoperative pain management, providing pain relief with few adverse effects. In this study, the effect of a preemptive regime on rat behavior and c-fos expression in the spinal cord of the uterine surgical pain model was evaluated. It was a lab-based experimental study in which 60 female Sprague-Dawley rats; eight to 10 weeks old, weighing 150-300 gm were used. The rats were divided into two main groups: (i) superficial pain group (SG) (with skin incision only), (ii) deep pain group (with skin and uterine incisions). Each group was further divided into three subgroups based on the type of preemptive analgesia administered i.e., "tramadol, buprenorphine, and saline subgroups." Pain behavior was evaluated using the "Rat Grimace Scale" (RGS) at 2, 4, 6, 9 and 24 h post-surgery. Additionally, c-fos immunohistochemistry was performed on sections from spinal dorsal horn (T12-L2), and its expression was evaluated using optical density and mean cell count 2hours postoperatively. Significant reduction in the RGS was noted in both the superficial and deep pain groups within the tramadol and buprenorphine subgroups when compared to the saline subgroup (p ≤ .05). There was a significant decrease in c-fos expression both in terms of number of c-fos positive cells and the optical density across the superficial laminae and lamina X of the spinal dorsal horn in both SD and DG (p ≤ .05). In contrast, the saline group exhibited c-fos expression primarily in laminae I-II and III-IV for both superficial and deep pain groups and lamina X in the deep pain group only (p ≤ .05). Hence, a preemptive regimen results in significant suppression of both superficial and deep components of pain transmission. These findings provide compelling evidence of the analgesic efficacy of preemptive treatment in alleviating pain response associated with uterine surgery.

Characteristics and exploration targets of Chang 7 shale oil in Triassic Yanchang Formation, Ordos Basin, NW China

The geological characteristics and enrichment laws of the shale oil in the third submember of the seventh member of Triassic Yanchang Formation (Chang 73) in the Ordos Basin were analyzed by using the information of core observations, experiments and logging, and then the exploration potential and orientation of the Chang 73 shale oil were discussed. The research findings are obtained in three aspects. First, two types of shale oil, i.e. migratory-retained and retained, are recognized in Chang 73. The former is slightly better than the latter in quality. The migratory-retained shale oil reservoir is featured with the frequent interbedding and overlapping of silty–sandy laminae caused by sandy debris flow and low-density turbidity current and semi-deep–deep lacustrine organic-rich shale laminae. The retained shale oil reservoir is composed of black shale with frequent occurrence of bedding and micro-laminae. Second, high-quality source rocks provide a large quantity of hydrocarbon-rich high-quality fluids with high potential energy. The source-reservoir pressure difference provides power for oil accumulation in thin interbeds of organic-poor sandstones with good seepage conditions and in felsic lamina, tuffaceous lamina and bedding fractures in shales. Hydrocarbon generation-induced fractures, bedding fractures and microfractures provide high-speed pathways for oil micro-migration. Frequent sandstone interlayers and felsic laminae provide a good space for large-scale hydrocarbon accumulation, and also effectively improve the hydrocarbon movability. Third, sand-rich areas around the depression are the main targets for exploring migratory-retained shale oil. Mature deep depression areas are the main targets for exploring retained oil with medium to high maturity. Theoretical research and field application of in-situ conversion in low-mature deep depression areas are the main technical orientations for exploring retained shale oil with low to medium maturity.

Dual contribution of ASIC1a channels in the spinal processing of pain information by deep projection neurons revealed by computational modeling

Dorsal horn of the spinal cord is an important crossroad of pain neuraxis, especially for the neuronal plasticity mechanisms that can lead to chronic pain states. Windup is a well-known spinal pain facilitation process initially described several decades ago, but its exact mechanism is still not fully understood. Here, we combine both ex vivo and in vivo electrophysiological recordings of rat spinal neurons with computational modeling to demonstrate a role for ASIC1a-containing channels in the windup process. Spinal application of the ASIC1a inhibitory venom peptides mambalgin-1 and psalmotoxin-1 (PcTx1) significantly reduces the ability of deep wide dynamic range (WDR) neurons to develop windup in vivo. All deep WDR-like neurons recorded from spinal slices exhibit an ASIC current with biophysical and pharmacological characteristics consistent with functional expression of ASIC1a homomeric channels. A computational model of WDR neuron supplemented with different ASIC1a channel parameters accurately reproduces the experimental data, further supporting a positive contribution of these channels to windup. It also predicts a calcium-dependent windup decrease for elevated ASIC conductances, a phenomenon that was experimentally validated using the Texas coral snake ASIC-activating toxin (MitTx) and calcium-activated potassium channel inhibitory peptides (apamin and iberiotoxin). This study supports a dual contribution to windup of calcium permeable ASIC1a channels in deep laminae projecting neurons, promoting it upon moderate channel activity, but ultimately leading to calcium-dependent windup inhibition associated to potassium channels when activity increases.

MP63-08 CLINICAL AND GENOMIC RISK FEATURES OF HIGH GRADE T1 NON-MUSCLE INVASIVE BLADDER CANCER

You have accessJournal of UrologyCME1 Apr 2023MP63-08 CLINICAL AND GENOMIC RISK FEATURES OF HIGH GRADE T1 NON-MUSCLE INVASIVE BLADDER CANCER Carissa Chu, Jessica Chen, Manuel DE Jesus Escano, Karissa Whiting, Irina Ostrovnaya, Song Jiang, Wesley Yip, Sari Khaleel, Andrew Lenis, Timothy Clinton, Goh Alvin, CHA Eugene, Timothy Donahue, Guido Dalbagni, Gopa Iyer, David Solit, Judy Sarungbam, Hikmat AL-Ahmadie, Bernard Bochner, and Eugene Pietzak Carissa ChuCarissa Chu More articles by this author , Jessica ChenJessica Chen More articles by this author , Manuel DE Jesus EscanoManuel DE Jesus Escano More articles by this author , Karissa WhitingKarissa Whiting More articles by this author , Irina OstrovnayaIrina Ostrovnaya More articles by this author , Song JiangSong Jiang More articles by this author , Wesley YipWesley Yip More articles by this author , Sari KhaleelSari Khaleel More articles by this author , Andrew LenisAndrew Lenis More articles by this author , Timothy ClintonTimothy Clinton More articles by this author , Goh AlvinGoh Alvin More articles by this author , CHA EugeneCHA Eugene More articles by this author , Timothy DonahueTimothy Donahue More articles by this author , Guido DalbagniGuido Dalbagni More articles by this author , Gopa IyerGopa Iyer More articles by this author , David SolitDavid Solit More articles by this author , Judy SarungbamJudy Sarungbam More articles by this author , Hikmat AL-AhmadieHikmat AL-Ahmadie More articles by this author , Bernard BochnerBernard Bochner More articles by this author , and Eugene PietzakEugene Pietzak More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003321.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: High grade non-muscle invasive bladder cancer invading the lamina propria (HG T1) exhibits variable clinical behavior. Existing risk stratification approaches by clinicopathologic alone are limited in determining who will benefit most from immediate radical cystectomy (RC) versus BCG. We report contemporary outcomes of patients managed with BCG or immediate RC and comparative mutational landscape across the spectrum of HG T1 disease. METHODS: This was a single-center retrospective cohort study of patients newly diagnosed with HG T1 non-muscle invasive bladder cancer between 2014 and 2020>18 months of follow up. All included patients underwent restaging TURBT with confirmed T1 by TURBT/EUA/imaging. Targeted exome sequencing with MSK-IMPACT was performed on 139 HG T1 tumors and 191 T2 tumors. RESULTS: 390 patients were included with median follow up time of 35 months (IQR 19-59 months). 308 were managed with intravesical BCG and 82 immediate RC. Patients selected for immediate RC were younger (median age 66 vs 69, p<0.001) and had higher rates of concurrent CIS (78 vs 53%, p<0.001), deep lamina propria invasion (37 vs 17%, p<0.001), lymphovascular invasion (7 vs 0.3%, p<0.001), and larger tumor size (4.3 cm vs 3.3 cm, p=0.007). Of the 82 patients who immediate radical cystectomy (BCG naïve), 32 (40%) were upstaged to pT2 and/or N+disease at RC. Persistent pT1 disease at re-resection and the presence of variant histology were associated with pathologic upstaging (p<0.01), while tumor size, concurrent CIS, bladder neck involvement, or prostatic urethra involvement were not. Compared to patients without additional risk factors, patients with CIS and at least one additional risk (e.g . “very high risk” T1 disease) harbored more TP53, ERBB2/Her2, and RB1 mutations and fewer FGFR3 mutations and a genomic profile more similar to T2 tumors than NMIBC (Figure 1). CONCLUSIONS: Very high risk T1 tumors are genomically similar to muscle-invasive tumors and are enriched in targetable alterations including ERBB2/Her2. Integration of genomic sequencing with clinical and pathologic features of HG T1 disease may improve risk stratification and treatment selection. Source of Funding: Ruth L. Kirschstein National Research Service Award T32CA082088 American Society of Clinical Oncology Young Investigator Award (YIA) Award © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e874 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Carissa Chu More articles by this author Jessica Chen More articles by this author Manuel DE Jesus Escano More articles by this author Karissa Whiting More articles by this author Irina Ostrovnaya More articles by this author Song Jiang More articles by this author Wesley Yip More articles by this author Sari Khaleel More articles by this author Andrew Lenis More articles by this author Timothy Clinton More articles by this author Goh Alvin More articles by this author CHA Eugene More articles by this author Timothy Donahue More articles by this author Guido Dalbagni More articles by this author Gopa Iyer More articles by this author David Solit More articles by this author Judy Sarungbam More articles by this author Hikmat AL-Ahmadie More articles by this author Bernard Bochner More articles by this author Eugene Pietzak More articles by this author Expand All Advertisement PDF downloadLoading ...

Ectopic expression of Nav1.7 in spinal dorsal horn neurons induced by NGF contributes to neuropathic pain in a mouse spinal cord injury model.

Neuropathic pain (NP) induced by spinal cord injury (SCI) often causes long-term disturbance for patients, but the mechanisms behind remains unclear. Here, our study showed SCI-induced ectopic expression of Nav1.7 in abundant neurons located in deep and superficial laminae layers of the spinal dorsal horn (SDH) and upregulation of Nav1.7 expression in dorsal root ganglion (DRG) neurons in mice. Pharmacologic studies demonstrated that the efficacy of the blood-brain-barrier (BBB) permeable Nav1.7 inhibitor GNE-0439 for attenuation of NP in SCI mice was significantly better than that of the BBB non-permeable Nav1.7 inhibitor PF-05089771. Moreover, more than 20% of Nav1.7-expressing SDH neurons in SCI mice were activated to express FOS when there were no external stimuli, suggesting that the ectopic expression of Nav1.7 made SDH neurons hypersensitive and Nav1.7-expressing SDH neurons participated in central sensitization and in spontaneous pain and/or walking-evoked mechanical pain. Further investigation showed that NGF, a strong activator of Nav1.7 expression, and its downstream JUN were upregulated after SCI in SDH neurons with similar distribution patterns and in DRG neurons too. In conclusion, our findings showed that the upregulation of Nav1.7 was induced by SCI in both SDH and DRG neurons through increased expression of NGF/JUN, and the inhibition of Nav1.7 in both peripheral and spinal neurons alleviated mechanical pain in SCI mice. These data suggest that BBB permeable Nav1.7 blockers might relieve NP in patients with SCI and that blocking the upregulation of Nav1.7 in the early stage of SCI via selective inhibition of the downstream signaling pathways of NGF or Nav1.7-targeted RNA drugs could be a strategy for therapy of SCI-induced NP.

Gastric endocrine cell hyperplasia after neoadjuvant therapy for poorly cohesive carcinoma: a diagnostic pitfall

Mucosal endocrine cell hyperplasia is a poorly described phenomenon that may occur following neoadjuvant therapy in a variety of gastrointestinal cancers. We describe two cases of poorly cohesive gastric carcinoma which underwent neoadjuvant chemotherapy. Subsequent biopsy and resection showed the presence of endocrine cells occurring singly and in micronests within the deep lamina propria, showing typical reactivity with synaptophysin and chromogranin. The distribution of these cells and cytokeratin positivity may mimic residual poorly cohesive carcinoma, particularly in small post-therapy biopsies. This phenomenon is thought to be related to normal mucosal elements being rendered relatively conspicuous by the depletion of malignant cells in cases with good neoadjuvant response. This treacherous finding may be misdiagnosed on small biopsies as residual malignancy, leading to potentially avoidable differences in subsequent management. This error may be avoided by awareness of this phenomenon and applying a simple panel of directed immunohistochemical markers, coupled with morphological correlation. Endocrine cell hyperplasia post neoadjuvant therapy in poorly cohesive carcinoma is a mimic of residual disease and may be result in false-positive interpretation of residual disease. Interpretation of post-treatment small biopsies should take into account this finding to avoid this potential clinically significant diagnostic pitfall.

Assessing the reliability of modern marine stromatolites as archives for the uranium isotope paleoredox proxy

Uranium isotopes (238U/235U) are commonly utilised in marine carbonates to reconstruct the redox evolution of ancient oceans and the atmosphere. However, questions remain concerning U isotope fractionation during carbonate precipitation and/or early sedimentary diagenesis. Moreover, the stromatolitic nature of many Precambrian limestones and dolostones brings further questions about U isotope fractionation during stromatolite formation due to microbial activity. Thus, it is unclear whether stromatolites are reliable archives for the U isotope composition of seawater. Here we measure the 238U/235U ratio of modern stromatolites from the hypersaline Shark Bay, Western Australia, a key study area that represents the largest modern example of stromatolite growth globally. A component-specific approach was utilised to analyse the uppermost crusts of modern stromatolites, older material from the deeper stromatolite laminae and the substrate upon which the stromatolites are forming. Our interpretations of δ238U data are aided by δ234U, δ18O, δ13C and trace element measurements, including some mineralogy and total organic carbon determinations for selected samples.Modern aragonitic crusts of stromatolites from the subtidal and intertidal zones around Shark Bay exhibited a narrow range of δ238U (−0.30 to −0.33‰), corresponding to a small offset of ca. +0.07‰ from seawater. This range of δ238U values overlaps with that of other primary carbonate precipitates in shallow marine environments such as corals but with a slightly higher mean δ238U. We investigated if this offset may be associated with the coprecipitation of U with aragonite from hypersaline seawater in Shark Bay, which has a higher proportion of dissolved U present as Ca2UO2(CO3)3 compared to open seawater. However, this measured offset is much smaller than that predicted using a speciation-dependent isotope fractionation model. Sub-recent material in the deeper stromatolite laminae exhibited higher and more variable δ238U (up to −0.15‰) compared to the modern aragonitic crusts of stromatolites. The strong inverse correlation between δ238U and the total organic carbon content (r = −0.83) and positive correlation with Mn concentrations (r = 0.91) suggest that 238U was preferentially reduced in the deeper laminae in association with organic carbon remineralisation under reducing conditions. The average δ238U offset of sub-recent stromatolite laminae from modern seawater was +0.13 ± 0.11‰ (1 s.d.), which is slightly lower than, but within uncertainty of, the offset measured for bulk carbonates from the Bahamas (+0.27 ± 0.14‰; 1 s.d.). These results demonstrate that stromatolites are valuable archives of global seawater δ238U, even in hypersaline restricted basins such as Shark Bay, but an offset to account for syndepositional diagenesis is likely required to accurately reconstruct seawater δ238U using ancient stromatolites.

Dorsal horn volume loss and pain pathway changes in Cavalier King Charles Spaniels with syringomyelia, signs of pain, and phantom scratching.

Central neuropathic pain is a core clinical sign of syringomyelia in humans and Cavalier King Charles Spaniel (CKCS) dogs. This histopathological study used spinal cords from CKCS dogs with syringomyelia to investigate the following conditions: (1) whether specific structural cervical spinal cord entities involved in nociception were affected by loss of neuroparenchyma or other pathological changes in CKCS dogs with pain-related behaviour and phantom scratching, (2) whether pain-related behaviour or phantom scratching correlated with loss of a specific anatomical entity or upregulation of glia cells, and (3) whether syringomyelia-related lesions affected specific functional spinal cord units of nociception. Spinal cord segments C1-C8 from CKCS dogs with magnetic resonance imaging-confirmed syringomyelia and clinical signs of pain and phantom scratching (n = 8) were compared with those from CKCS dogs without syringomyelia (n = 4). Dogs with unilateral scratching (n = 7) had a volume loss ( P = 0.043) of the dorsal horn laminae I-III in the ipsilateral side compared with the contralateral dorsal horn. A clear pattern of ipsilateral changes in the dorsal root entry zone characterised by deafferentation and reorganization of first-order axons into deeper laminae was found in cases with lateralised scratching. Significant changes in cell number density were not found for astrocytes or microglia, suggesting that the dogs represented cases of end-stage syringomyelia and thus could not reveal astrogliosis and microgliosis, which may be involved in the early phases of syrinx development and phantom scratching. The present relationship between clinical findings and dorsal horn and pain pathway pathology in CKCS dogs suggests that these dogs may be of interest as a supplement to experimental model pain research.

Ultrasound Analyses of the Dorsal Hands for Volumetric Rejuvenation

Volumetric hand rejuvenation of the dorsal hands is increasingly applied for aged, skeletonized dorsal hand skin, with prominent veins, tendons, and joints due to volumetric loss of subcutaneous fat. However, the aging process of the fatty laminae remains unexplored. The aim of this study was to investigate the impact of aging and potential determinants on the fatty laminae to improve the therapeutic effect of volumetric injection. This cross-sectional study enrolled 105 Taiwanese participants aged ≥20 years, divided into 5 decade-based age groups. A single blinded practitioner measured the thicknesses of the dorsal superficial lamina (DSL), dorsal intermediate lamina (DIL), and dorsal deep lamina (DDL) by ultrasonography. The potential determining factors of the laminae thickness (age, sex, BMI, and hand dominance) were analyzed. The thicknesses of the 3 laminae decreased with age, with the mean decrease in thickness from the 20s to >60 years of the DSL, DIL, and DDL being 0.21 mm (30.0%), 1.38 mm (63.89%), and 0.31 mm (20.95%), respectively. The decrease in DIL thickness was the greatest and most significant in subjects aged >30 years. Multiple linear regression analysis showed age to be the only determinant of thickness for the 3 laminae (all P < 0.001), although the DIL was significantly thicker in men (P < 0.001). Volumetric fat loss was noted in the 3 fatty laminae of dorsal hands during aging; the DIL showed the greatest progressive fat loss after the age of 30. Volumetric rejuvenation of the 3 laminae may result in the most aesthetic appearance, especially in women.

Predicting Wide-Dynamic Range Neuron Activity from Peripheral Nerve Stimulation using Linear Parameter Varying Models.

Neuromodulation treatments for chronic pain are programmed with limited knowledge of how electrical stimulation of nerve fibers affects the dynamic response of pain-processing neurons in the spinal cord and the brain. By modeling these effects with tractable representations, we may be able to improve efficacy of stimulation therapy. However, pain transmitting neurons in the dorsal horn of the spinal cord, the first pain relay station in the nervous system, have complex responses to peripheral nerve stimulation (PNS) with nonlinearities and history effects. Wide-dynamic range (WDR) neurons are well studied in pain models and respond to peripheral noxious and non-noxious stimuli. We propose to use linear parameter varying (LPV) models to capture PNS responses of WDR neurons of the deep lamina in the dorsal horn in the spinal cord. Here we show that LPV models perform better than a single linear time-invariant (LTI) model in representing the responses of the WDR neurons to widely varying amplitudes of PNS current. In the future, we can use these models alongside LPV control techniques to design closed-loop PNS stimulation that may accomplish optimal pain treatment goals.Clinical Relevance- Electrical nerve stimulation as a therapy for chronic pain is in need of a more informed approach to programming. By describing the effects of stimulation on the pain system with tractable mathematical models, we may be able to titrate the stimulation to more effectively treat chronic pain.

Spinal Cord Stimulation Reduces Ventricular Arrhythmias by Attenuating Reactive Gliosis and Activation of Spinal Interneurons.

This study investigated spinal cord neuronal and glial cell activation during cardiac ischemia-reperfusion (IR)-triggered ventricular arrhythmias and neuromodulation therapy by spinal cord stimulation (SCS). Myocardial ischemia induces changes in cardiospinal neural networks leading to sudden cardiac death. Neuromodulation with SCS decreases cardiac sympathoexcitation; however, the molecular mechanisms remain unknown. Yorkshire pigs (n=16) were randomized to Control, IR, or IR+SCS groups. A 4-pole SCS lead was placed in the T1-T4 epidural space with stimulation for 30minutes before IR (50Hz, 0.4-ms duration, 90% motor threshold). Cardiac electrophysiological mapping and Ventricular Arrhythmia Score (VAS) were recorded. Immunohistochemistry of thoracic spinal sections was used to map and identify Fos-positive neuronal and glial cell types during IR with and without SCS. IR increased cardiac sympathoexcitation and arrhythmias (VAS=6.2 ± 0.9) that were attenuated in IR+ SCS (VAS=2.8 ± 0.5; P=0.017). IR increased spinal cellular Fos expression (#Fos+ cells Control=23 ± 2 vs IR=88 ± 5; P< 0.0001) in T1-T4, with the greatest increase localized to T3, and the greatest %Fos+ cells being microglia and astrocytes. Fos expression was attenuated by IR+ SCS (62 ± 4; P< 0.01), primarily though a reduction in Fos+ microglia and astrocytes, as SCS also led to increase in Fos+ neurons in deep dorsal laminae. In a porcine model, cardiac IR was associated with astrocyte and microglial cell activation. Our results suggest that preemptive thoracic SCS decreased IR-induced cardiac sympathoexcitation and ventricular arrhythmias through attenuation of reactive gliosis and activation of inhibitory interneurons in the dorsal horn of spinal cord.

Leukocyte numbers and intestinal mucosal morphometrics in horses with no clinical intestinal disease.

Healthy horses and other animals have large numbers of resident leukocytes in the intestinal wall, but there is scant information regarding which and how many leukocytes are normally present in the equine intestinal wall. Our aim was to provide a reference range of leukocytes in the intestinal mucosal and submucosal propria of normal horses. We included in our study intestinal tissues from 22 Thoroughbred racehorses with no clinical intestinal disease, which had been euthanized because of catastrophic musculoskeletal injuries. Neutrophils, lymphocytes, eosinophils, macrophages, and plasma cells were counted in 5 random 17,600-µm2 areas of villus lamina propria of the duodenum, jejunum, and ileum, and deep lamina propria of the duodenum, jejunum, ileum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon, and small colon. Other features investigated in the same intestinal segments included villus height and width (small intestine), presence of ciliated protozoa, Paneth cells number, subcryptal leukocyte layers (number of leukocyte layers between the bottom of the crypts and the muscularis mucosae), and submucosal leukocytes. Lymphocytes were the most numerous cells in all segments analyzed, followed by plasma cells, eosinophils, macrophages, and neutrophils. Eosinophil numbers were significantly higher in both lamina propria and submucosa of the large intestine than in the small intestine. The duodenum had shorter and thinner villi than either jejunum or ileum. The data provided from our study will be useful for diagnosticians examining inflammatory processes in the intestinal tract of horses.

Histopathological analysis of the association between mucosal epithelial changes and the lamina propria vascular network in irritation fibroma

ObjectivesIrritation fibroma in the oral cavity causes atrophy or squamous epithelium thickening with respect to external injury-associated factors. However, ulcers do not occur in most cases. This study aimed to elucidate the mechanism by which ulcers do not form, focusing on the vascular network in the mucosal epithelium of irritation fibroma. MethodsImmunostaining was performed using an enzyme antibody method with primary antibodies against CD31 and Ki-67 in 17 cases of irritation fibroma in the buccal mucosa. One section was taken at three points from the margin and three points from just above the lesion for measurement. The number of blood vessels in the superficial and deep lamina propria at the measurement site were determined, and the area per blood vessel was measured. ResultsThe number and area of blood vessels in the superficial lamina propria just below the lesion epithelium were smaller than those in the margin. No difference was observed in the number and area of blood vessels in the deep lamina propria between the margins and lesions. ConclusionsOur results suggest that the vascular network in the deep lamina propria is maintained and compensates for the nutrient supply to the covering epithelium.

Clinical Characteristics and Outcomes of Eosinophilic Colitis in Patients With Cancer.

Eosinophilic colitis (EoC) is a rare form of eosinophilic gastrointestinal disease characterized by diffuse eosinophilic infiltration in the deep lamina propria of colonic mucosa. The pathophysiology is unclear, but EoC has been associated with multiple known risk factors. The aim of this study was to characterize the clinical characteristics and disease course of patients with EoC at a major cancer center. We retrospectively reviewed colonic samples obtained between January 2000 and December 2018 from our institutional database and included cases with significant colonic eosinophilia. Baseline clinical data and EoC-related clinical course and outcomes were documented. Forty-one patients were included. One fourth had coexisting autoimmune conditions. Seventy-eight percent had a cancer diagnosis. Half the patients received chemotherapy, with a median duration of 180 days between chemotherapy and EoC onset. Symptoms were present in 76% of patients. Diarrhea was more prevalent in patients who received chemotherapy (85% vs. 42%). Median duration of EoC symptoms was 30 days in patients with cancer and 240 days in those without cancer (P=0.03). Most patients (88%) had normal colonoscopy findings. Fifteen percent of patients required hospitalization. All-cause mortality was 37%, mostly related to underlying malignancy and organ failure. EoC in cancer patients appears to have more diarrhea-predominant symptoms, particularly in patients receiving chemotherapy, but a shorter disease duration compared with patients without cancer. Hospitalization can be required for serious cases. Treatment may be reserved for patients requiring symptom management, as most patients with EoC have good clinical outcomes regardless of treatment.

Spinoparabrachial projection neurons form distinct classes in the mouse dorsal horn.

Projection neurons in the spinal dorsal horn relay sensory information to higher brain centres. The activation of these populations is shaped by afferent input from the periphery, descending input from the brain, and input from local interneuron circuits. Much of our recent understanding of dorsal horn circuitry comes from studies in transgenic mice; however, information on projection neurons is still based largely on studies in monkey, cat, and rat. We used viral labelling to identify and record from mouse parabrachial nucleus (PBN) projecting neurons located in the dorsal horn of spinal cord slices. Overall, mouse lamina I spinoparabrachial projection neurons (SPBNs) exhibit many electrophysiological and morphological features that overlap with rat. Unbiased cluster analysis distinguished 4 distinct subpopulations of lamina I SPBNs, based on their electrophysiological properties that may underlie different sensory signalling features in each group. We also provide novel information on SPBNs in the deeper lamina (III-V), which have not been previously studied by patch clamp analysis. These neurons exhibited higher action potential discharge frequencies and received weaker excitatory synaptic input than lamina I SPBNs, suggesting this deeper population produces different sensory codes destined for the PBN. Mouse SPBNs from both regions (laminae I and III-V) were often seen to give off local axon collaterals, and we provide neuroanatomical evidence they contribute to excitatory input to dorsal horn circuits. These data provide novel information to implicate excitatory input from parabrachial projection neuron in dorsal horn circuit activity during processing of nociceptive information, as well as defining deep dorsal horn projection neurons that provide an alternative route by which sensory information can reach the PBN.