Intact Signals Research Articles

Deep LSTM-based autoencoder for CFRP damage imaging using Lamb wave and correlation factor weighted delay-and-sum algorithm

The ultrasonic Lamb wave testing (ULWT) has proven valuable in non-destructive testing (NDT) due to its high sensitivity and wide coverage. However, the classical post-processing algorithm, the delay-and-sum (DAS) technique, suffers from notable artifacts and inadequate accuracy in Lamb wave inspection due to the existence of reflection and superposition, especially on composite materials. In this study, we proposed a novel algorithm, the correlation factor weighted DAS imaging algorithm based on the Long short-term memory-autoencoder (LSTM-AE), to address these deterioration issues. The LSTM-AE demonstrates the capability to extract potential features and accurately reconstruct input signals. By incorporating the concept of anomaly detection, an LSTM-AE trained with intact signals produces a significantly distorted output when exposed to damaged signals as input. We formulated a novel damage index (DI) based on the Pearson Correlation Coefficient (PCC) between the output and input signals as the weighting factor. The proposed method has undergone experimental validation, confirming its effectiveness in Lamb wave inspection.

Uterine Inflammatory Myofibroblastic Tumor with ALK Polysomy: Challenging Case Report and Review of Literature

Abstract Introduction/Objective Inflammatory myofibroblastic tumor (IMT) is a distinct neoplastic process with specific molecular alterations. Given its low likelihood of local recurrence and metastasis, it is currently regarded as a neoplasm with intermediate biological potential. IMTs of the uterus variably express CD10 and smooth muscle markers, which present a diagnostic challenge and make it difficult to distinguish from mesenchymal neoplasms of smooth muscle and endometrial stromal origin. Methods/Case Report We present a case of a 46-year-old woman who underwent a hysterectomy due to abnormal menstrual bleeding. Grossly, a 5.0 cm tan, firm, well-circumscribed subendometrial nodule was identified at the fundus, which had a fleshy, soft-cut surface with no hemorrhage or necrosis. Microscopically, the nodule is composed of somewhat tightly packed spindle cells arranged in a fascicular pattern, with little-to-no myxoid stroma and dense lymphoplasmacytic infiltrate forming scattered aggregates. Immunohistochemical stains show lesional spindle cells are positive for SMA, desmin, and vimentin with patchy staining for ER while negative for S100, CD10, ALK-1, STAT6, and CD34. CD68 highlights scattered histiocytes in the background. Ki-67 highlights background lymphoid cells. RNA next-generation sequencing detected no fusions involving ROS1, PDGFR-β, or ETV6, which were previously described in a subset of ALK-negative IMTs. The FISH assay did not reveal rearrangements of the ALK or ROS1. However, extra intact ALK fusion signals were observed, suggesting polysomy for the ALK region. Results (if a Case Study enter NA) NA Conclusion To our knowledge, this is the first report of a uterine IMT with ALK polysomy. The significance of additional ALK copies in our case is not clear but highlights the need for further molecular studies investigating genetic alterations in ALK-negative IMTs. Detecting these rearrangements may be of diagnostic value and help guide future targeted therapies.

O026 Concordance between a home sleep apnoea test based on peripheral artery tonometry and laboratory polysomnography

Abstract Introduction Limited channel home sleep apnoea test (HSAT) devices are an emerging alternative to laboratory-based PSG. Despite increasingly commercially availability, there is limited external validation of their diagnostic performance. We recently independently validated the Night Owl Mini HSAT (NOM) against in-lab PSG and demonstrated a satisfactory level of agreement (LOA) of 63.8% overall. However, lower rates of agreement were observed for severe OSA. We speculated that signal artefact may have contributed to reduced concordance. Methods Data was drawn from our previously published prospective cohort of 100 participants undergoing PSG for suspected OSA, simultaneously fitted with the NOM. PSG and NOM studies were re-processed, manually remarking artefact. LOA was recalculated comparing only temporally aligned data segments for which both devices provided intact signals, this allowed direct comparison of pulse oximetry data and software algorithms that generated the ODI3%. Two new datasets were generated for PSG and NOM for which combined artefact was deleted (COM-PSG, COM-NOM). Results To date, datasets from 12 participants have been assessed. The mean difference in ODI3% between PSG and NOM was 5.25/hr (95% limits of agreement -0.73 – 11.23), while the mean difference in ODI3% between COM-PSG and COM-NOM was 2.53/hr (95% limits of agreement -1.77 – 6.83). In our previous study, the overall mean difference in ODI3% was –0.21/hr (95% limits of agreement -18.1 – 17.7). Conclusion Preliminary results may indicate signal artefact contributes to reduced level of agreement between PSG and NOM.

Improvement of Lipid Detection in Mouse Brain and Human Uterine Tissue Sections Using In Situ Matrix Enhanced Secondary Ion Mass Spectrometry.

The potential of mass spectrometry imaging, and especially ToF-SIMS 2D and 3D imaging, for submicrometer-scale, label-free molecular localization in biological tissues is undisputable. Nevertheless, sensitivity issues remain, especially when one wants to achieve the best lateral and vertical (nanometer-scale) resolution. In this study, the interest of in situ matrix transfer for tissue analysis with cluster ion beams (Bin+, Arn+) is explored in detail, using a series of six low molecular weight acidic (MALDI) matrices. After estimating the sensitivity enhancements for phosphatidylcholine (PC), an abundant lipid type present in almost any kind of cell membrane, the most promising matrices were softly transferred in situ on mouse brain and human uterine tissue samples using a 10 keV Ar3000+ cluster beam. Signal enhancements up to 1 order of magnitude for intact lipid signals were observed in both tissues under Bi5+ and Ar3000+ bombardment. The main findings of this study lie in the in-depth characterization of uterine tissue samples, the demonstration that the transferred matrices also improve signal efficiency in the negative ion polarity and that they perform as well when using Bin+ and Arn+ primary ions for analysis and imaging.

O-166 Distinct molecular signatures of successful sperm in situ

Abstract Out of millions of ejaculated sperm, a few reach the fertilization site in mammals. Flagellar Ca2+ signaling nanodomains, organized by multi-subunit CatSper calcium channel complexes, are pivotal for sperm migration in the female tract, implicating CatSper-dependent mechanisms in sperm selection. Here using biochemical and pharmacological studies, we demonstrate that CatSper1 is an O-linked glycosylated protein, undergoing capacitation-induced processing dependent on Ca2+ and phosphorylation cascades. CatSper1 processing correlates with protein tyrosine phosphorylation (pY) development in sperm cells capacitated in vitro and in vivo. Using 3D in situ molecular imaging and ANN-based automatic detection of sperm distributed along the cleared female tract, we demonstrate that spermatozoa past the utero-tubal junction possess the intact CatSper1 signals. Together, we reveal that fertilizing mouse spermatozoa in situ are characterized by intact CatSper channel, lack of pY, and reacted acrosomes. These findings provide molecular insight into sperm selection for successful fertilization in the female reproductive tract.

Chemical Decrosslinking-Based Peptide Characterization of Formaldehyde-Fixed Rat Pancreas Using Fluorescence-Guided Single-Cell Mass Spectrometry.

Approaches for the characterization of proteins/peptides in single cells of formaldehyde-fixed (FF) tissues via mass spectrometry (MS) are still under development. The lack of a general method for selectively eliminating formaldehyde-induced crosslinking is a major challenge. A workflow is shown for the high-throughput peptide profiling of single cells isolated from FF tissues, here the rodent pancreas, which possesses multiple peptide hormones from the islets of Langerhans. The heat treatment is enhanced by a collagen-selective multistep thermal process assisting efficient isolation of islets from the FF pancreas and, subsequently, their dissociation into single islet cells. Hydroxylamine-based chemical decrosslinking helped restore intact peptide signals from individual isolated cells. Subsequently, an acetone/glycerol-assisted cell dispersion was optimized for spatially resolved cell deposition onto glass slides, while a glycerol solution maintained the hydrated state of the cells. This sample preparation procedure allowed peptide profiling in FF single cells by fluorescence-guided matrix-assisted laser desorption ionization MS. Here, 2594 single islet cells were analyzed and 28 peptides were detected, including insulin C-peptides and glucagon. T-distributed stochastic neighbor embedding (t-SNE) data visualization demonstrated that cells cluster based on cell-specific pancreatic peptide hormones. This workflow expands the sample availability for single-cell MS characterization to a wide range of formaldehyde-treated tissue specimens stored in biobanks.

Sorted compressive sensing for reconstruction of failed in-core detector signals

Self-Powered Neutron Detectors(SPNDs) are used to calculate core power distributions, an essential factor in the safe operation of nuclear power plants. Some detectors may fail during normal operation, and signals from failed detectors are isolated from intact signals. The calculated detailed power distribution accuracy depends on the number of available detector signals. Failed detectors decrease the operating margin by enlarging the power distribution measurement error. Therefore, a thorough reconstruction of the failed detector signals is critical. This note suggests a compressive sensing based methodology that rationally reconstructs the readings of failed detectors. The methodology significantly improves reconstruction accuracy by sorting signals and removing high-frequency components from conventional compressive sensing methodology.

Psychological stress disrupts intestinal epithelial cell function and mucosal integrity through microbe and host-directed processes

ABSTRACT Psychological stress alters the gut microbiota and predisposes individuals to increased risk for enteric infections and chronic bowel conditions. Intestinal epithelial cells (IECs) are responsible for maintaining homeostatic interactions between the gut microbiota and its host. In this study, we hypothesized that disruption to colonic IECs is a key factor underlying stress-induced disturbances to intestinal homeostasis. Conventionally raised (CONV-R) and germ-free (GF) mice were exposed to a social disruption stressor (Str) to ascertain how stress modifies colonic IECs, the mucosal layer, and the gut microbiota. RNA sequencing of IECs isolated from CONV-R mice revealed a robust pro-inflammatory (Saa1, Il18), pro-oxidative (Duox2, Nos2), and antimicrobial (Reg3b/g) transcriptional profile as a result of Str. This response occurred concomitant to mucus layer thinning and signs of microbial translocation. In contrast to their CONV-R counterparts, IECs from GF mice or mice treated with broad spectrum antibiotics exhibited no detectable transcriptional changes in response to Str. Nevertheless, IECs from Str-exposed GF mice exhibited an altered response to ex vivo bacterial challenge (increased dual Oxidase-2 [Duox2] and nitric oxide synthase-2 (Nos2)), indicating that STR primes host IEC pro-oxidative responses. In CONV-R mice stress-induced increases in colonic Duox2 and Nos2 (ROS generating enzymes) strongly paralleled changes to microbiome composition and function, evidencing Str-mediated ROS production as a primary factor mediating gut-microbiota dysbiosis. In conclusion, a mouse model of social stress disrupts colonic epithelial and mucosal integrity, a response dependent on an intact microbiota and host stress signals. Together these preclinical findings may provide new insight into mechanisms of stress-associated bowel pathologies in humans.

The Muscle Cuff Regenerative Peripheral Nerve Interface for the Amplification of Intact Peripheral Nerve Signals.

Robotic exoskeletons have gained recent acclaim within the field of rehabilitative medicine as a promising modality for functional restoration for those individuals with extremity weakness. However, their use remains largely confined to research institutions, frequently operating as a means of static extremity support as motor detection methods remain unreliable. Peripheral nerve interfaces have arisen as a potential solution to this shortcoming; however, due to their inherently small amplitudes, these signals can be difficult to differentiate from background noise, lowering their overall motor detection accuracy. As current interfaces rely on abiotic materials, inherent material breakdown can occur alongside foreign body tissue reaction over time, further impacting their accuracy. The Muscle Cuff Regenerative Peripheral Nerve Interface (MC-RPNI) was designed to overcome these noted complications. Consisting of a segment of free muscle graft secured circumferentially to an intact peripheral nerve, the construct regenerates and becomes reinnervated by the contained nerve over time. In rats, this construct has demonstrated the ability to amplify a peripheral nerve's motor efferent action potentials up to 100 times the normal value through the generation of compound muscle action potentials (CMAPs). This signal amplification facilitates high accuracy detection of motor intent, potentially enabling reliable utilization of exoskeleton devices.

The Muscle Cuff Regenerative Peripheral Nerve Interface for the Amplification of Intact Peripheral Nerve Signals

The Muscle Cuff Regenerative Peripheral Nerve Interface for the Amplification of Intact Peripheral Nerve Signals

Effects of solvent composition on ionization and fragmentation within the solution-cathode glow discharge

Recently, solution-electrode glow discharges (SEGDs) have shown great utility as ionization sources for mass spectrometry (MS). The solution composition of SEGD electrodes is pivotal to their performance as it influences analyte-ion formation. The performance of electrospray ionization is heavily dictated by spray-solution composition, which can alter ionization efficiency and pathways. While SEGDs produce Taylor cones similar to electrospray ionization, the influence of solution-electrode composition on molecular-ion formation has not been studied. Here, we examine how additives to an atmospheric-pressure solution-cathode glow discharge (SCGD) influence molecular ionization and fragmentation. The impact of several additives to the acidic solution of an SCGD ionization source was evaluated based on mass-spectrometric performance. Addition of methanol increased molecular- and fragment-ion signals for peptide angiotensin II. This effect is likely due to improved desolvation and a greater interaction of analyte molecules with glow-discharge species. Several high-boiling-point reagents were tested to examine changes in the ion signal, the average charge state, and the degree of fragmentation. Overall, these additives inhibited fragmentation but significantly lowered intact molecular-ion signals. Interestingly, loss of fragment ions trended with the boiling point of the reagent used. We hypothesize that analyte molecules become trapped in droplets produced at the solution-cathode surface. These droplets do not fully desolvate before escaping the discharge region, sparing analyte molecules from fragmentation. For low volatility additives, droplets do not desolvate, even as they enter the heated MS, which yields a loss in molecular ions. The changing composition of the SCGD solution alters analytical performance, but also provides insight into analyte ionization and fragmentation processes.

Surface crack characterization using laser nonlinear ultrasonics based on the bispectrum

Laser nonlinear wave modulation spectroscopy (LNWMS) has been used to detect nonlinear ultrasonic signatures caused by fatigue cracks in materials. The nonlinear feature extraction from the spectral plot plays a significant role in LNWMS. This paper uses the bispectrum to distinguish harmonically related signals which result from nonlinear interactions and to remove those which do not. A feature called bispectrum damage factor (BDF), which is defined as the ratio of the bispectrum peak difference value of the damage and intact signals to the bispectrum peak value of intact signals at frequencies F 1, F 2, is extracted from the bispectral plot to characterize the crack features. The basic premise of the proposed laser nonlinear wave modulation bispectrum (LNWMBS) is that this BDF value will vary regularly as the level of crack-induced nonlinearity increases. A finite element model and a noncontact laser ultrasonic system have been built for LNWMBS measurement. The simulation results show that the BDF value is sensitive to crack depth and angle, and is insensitive to crack length and detecting position. BDF values of reflected Rayleigh waves are positive and approximately linearly increase with an increase of the crack depths in the range of 0.1 λ ∼ 0.6 λ (λ is the corresponding wavelength of adopted frequency); meanwhile, the values of the transmitted waves are negative and decreasing. The BDF values are larger than the crack angle at 90° when they are less than 90°, which is beneficial for crack detection. The proposed LNWMBS technique has been successfully tested for detecting artificial sub-millimeter surface-breaking cracks in aluminum alloy plates and closed fatigue cracks of torque load simulator connection blocks.

3D in situ imaging of the female reproductive tract reveals molecular signatures of fertilizing spermatozoa in mice.

Out of millions of ejaculated sperm, a few reach the fertilization site in mammals. Flagellar Ca2+ signaling nanodomains, organized by multi-subunit CatSper calcium channel complexes, are pivotal for sperm migration in the female tract, implicating CatSper-dependent mechanisms in sperm selection. Here using biochemical and pharmacological studies, we demonstrate that CatSper1 is an O-linked glycosylated protein, undergoing capacitation-induced processing dependent on Ca2+ and phosphorylation cascades. CatSper1 processing correlates with protein tyrosine phosphorylation (pY) development in sperm cells capacitated in vitro and in vivo. Using 3D in situ molecular imaging and ANN-based automatic detection of sperm distributed along the cleared female tract, we demonstrate that spermatozoa past the utero-tubal junction possess the intact CatSper1 signals. Together, we reveal that fertilizing mouse spermatozoa in situ are characterized by intact CatSper channel, lack of pY, and reacted acrosomes. These findings provide molecular insight into sperm selection for successful fertilization in the female reproductive tract.

Identification of eggshell crack for hen egg and duck egg using correlation analysis based on acoustic resonance method

AbstractThis work presents a novel method for eggshell crack timely detection of hen and duck eggs based on the acoustic resonance analysis. The developed experimental device consisting of an excitation device, an acquisition module of acoustic signals, and a personal computer was designed to generate and acquire the response signals by impaction on eggshell. The parameter of Pearson's correlation coefficients (PCCs) was calculated to compare the similarity between intact and crack eggs signals with various sampling points. The 200 sampling points of the response signal were selected as an effective response signal, which observed the values of higher than 0.85 and lower than 0.8 for intact eggs and crack eggs, respectively. Thereafter, multivariate analysis of variance (MANOVA) was conducted to find that the egg individual and the impact location were two remarkable influencing factors to the response signal from intact egg. The cross‐comparison of signals from the same direction of each egg sample was proposed to get the PCCs and served as feature parameters. A universal linear discriminant function was built to distinguish intact and crack egg. In the conducted experiments, a crack detection level of 95.5% and a false rejection level of 5% were achieved using the mixed samples of 100 hen eggs and 100 duck eggs. From the findings, it can be concluded that the proposed method will assist in simplifying the classification algorithm and in advancing the applicability for multiple categories of egg for an online detection system.Practical ApplicationsEggshell crack produced during packing and transportation may lead to significant economic loss and food security problem. This article presents a novel method of eggshell crack detection based on the acoustic resonance analysis for different categories egg. The effective response signals selection method and remarkable influencing factors analysis method of response signal was conducted to weaken the random interference of noise and reduce the computation consumption. As a result, a universal linear discriminant function was built to distinguish the intact and cracked eggs, which can satisfy the requirement in sorting industry. The overall results sufficiently indicated that the proposed methods in this study have significant potential for online detection of eggshell crack with high throughput.

Proteome dynamics from heavy water metabolic labeling and peptide tandem mass spectrometry

Protein homeostasis (proteostasis) is a result of a dynamic equilibrium between protein synthesis and degradation. It is important for healthy cell/organ functioning and is often associated with diseases such as neurodegenerative diseases and non-Alcoholic Fatty Liver disease. Heavy water metabolic labeling, combined with liquid-chromatography and mass spectrometry (LC-MS), is a powerful approach to study proteostasis in vivo in high throughput. Traditionally, intact peptide signals are used to estimate stable isotope incorporation in time-course experiments. The time-course of label incorporation is used to extract protein decay rate constant (DRC). Intact peptide signals, computed from integration in chromatographic time and mass-to-charge ratio (m/z) domains, usually, provide an accurate estimate of label incorporation. However, sample complexity (co-elution), limited dynamic range, and low signal-to-noise ratio (S/N) may adversely interfere with the peptide signals. These artifacts complicate the DRC estimations by distorting peak shape in chromatographic time and m/z domains. Fragment ions, on the other hand, are less prone to these artifacts and are potentially well suited in aiding DRC estimations. Here, we show that the label incorporation encoded into the isotope distributions of fragment ions reflect the isotope enrichment during the metabolic labeling with heavy water. We explore the label incorporation statistics for devising practical approaches for DRC estimations.

Chemical imaging of aggressive basal cell carcinoma using time-of-flight secondary ion mass spectrometry.

A set of basal cell carcinoma samples, removed by Mohs micrographic surgery and pathologically identified as having an aggressive subtype, have been analyzed using time-of-flight secondary ion mass spectrometry (SIMS). The SIMS analysis employed a gas cluster ion beam (GCIB) to increase the sensitivity of the technique for the detection of intact lipid species. The GCIB also allowed these intact molecular signals to be maintained while surface contamination and delocalized chemicals were removed from the upper tissue surface. Distinct mass spectral signals were detected from different regions of the tissue (epidermis, dermis, hair follicles, sebaceous glands, scar tissue, and cancerous tissue) allowing mass spectral pathology to be performed. The cancerous regions of the tissue showed a particular increase in sphingomyelin signals that were detected in both positive and negative ion mode along with increased specific phosphatidylserine and phosphatidylinositol signals observed in negative ion mode. Samples containing mixed more and less aggressive tumor regions showed increased phosphatidylcholine lipid content in the less aggressive areas similar to a punch biopsy sample of a nonaggressive nodular lesion.

Enhanced TH17 Responses in Patients with IL10 Receptor Deficiency and Infantile-onset IBD.

IL10 receptor (IL10R) deficiency causes severe infantile-onset inflammatory bowel disease. Intact IL10R-dependent signals have been shown to be important for innate and adaptive immune cell functions in mice. We have previously reported a key role of IL10 in the generation and function of human anti-inflammatory macrophages. Independent of innate immune cell defects, the aim of the current study was to determine the role of IL10R signaling in regulating human CD4 T-cell function. Peripheral blood mononuclear cells and intestinal biopsies cells were collected from IL10/IL10R-deficient patients and controls. Frequencies of CD4 T-cell subsets, naive T-cell proliferation, regulatory T cell (Treg)-mediated suppression, and Treg and TH17 generation were determined by flow cytometry. Transcriptional profiling was performed by NanoString and quantitative real-time polymerase chain reaction. RNA in situ hybridization was used to determine the quantities of various transcripts in intestinal mucosa. Analysis of 16 IL10- and IL10R-deficient patients demonstrated similar frequencies of peripheral blood and intestinal Tregs, compared with control subjects. In addition, in vitro Treg suppression of CD4 T-cell proliferation and generation of Treg were not dependent on IL10R signaling. However, IL10R-deficient T naive cells exhibited higher proliferative capacity, a strong TH17 signature, and an increase in polarization toward TH17 cells, compared with controls. Moreover, the frequency of TH17 cells was increased in the colon and ileum of IL10R-deficient patients. Finally, we show that stimulation of IL10R-deficient Tregs in the presence of IL1β leads to enhanced production of IL17A. IL10R signaling regulates TH17 polarization and T-cell proliferation in humans but is not required for the generation and in vitro suppression of Tregs. Therapies targeting the TH17 axis might be beneficial for IL10- and IL10R-deficient patients as a bridge to allogeneic hematopoietic stem cell transplantation.

Association of Neural and Emotional Impacts of Reward Prediction Errors With Major Depression

Major depressive disorder (MDD) is associated with deficits in representing reward prediction errors (RPEs), which are the difference between experienced and predicted reward. Reward prediction errors underlie learning of values in reinforcement learning models, are represented by phasic dopamine release, and are known to affect momentary mood. To combine functional neuroimaging, computational modeling, and smartphone-based large-scale data collection to test, in the absence of learning-related concerns, the hypothesis that depression attenuates the impact of RPEs. Functional magnetic resonance imaging (fMRI) data were collected on 32 individuals with moderate MDD and 20 control participants who performed a probabilistic reward task. A risky decision task with repeated happiness ratings as a measure of momentary mood was also tested in the laboratory in 74 participants and with a smartphone-based platform in 1833 participants. The study was conducted from November 20, 2012, to February 17, 2015. Blood oxygen level-dependent activity was measured in ventral striatum, a dopamine target area known to represent RPEs. Momentary mood was measured during risky decision making. Of the 52 fMRI participants (mean [SD] age, 34.0 [9.1] years), 30 (58%) were women and 32 had MDD. Of the 74 participants in the laboratory risky decision task (mean age, 34.2 [10.3] years), 44 (59%) were women and 54 had MDD. Of the smartphone group, 543 (30%) had a depression history and 1290 (70%) had no depression history; 918 (50%) were women, and 593 (32%) were younger than 30 years. Contrary to previous results in reinforcement learning tasks, individuals with moderate depression showed intact RPE signals in ventral striatum (z = 3.16; P = .002) that did not differ significantly from controls (z = 0.91; P = .36). Symptom severity correlated with baseline mood parameters in laboratory (ρ = -0.54; P < 1 × 10-6) and smartphone (ρ = -0.30; P < 1 × 10-39) data. However, participants with depression showed an intact association between RPEs and happiness in a computational model of momentary mood dynamics (z = 4.55; P < .001) that was not attenuated compared with controls (z = -0.42; P = .67). The neural and emotional impact of RPEs is intact in major depression. These results suggest that depression does not affect the expression of dopaminergic RPEs and that attenuated RPEs in previous reports may reflect downstream effects more closely related to aberrant behavior. The correlation between symptom severity and baseline mood parameters supports an association between depression and momentary mood fluctuations during cognitive tasks. These results demonstrate a potential for smartphones in large-scale computational phenotyping, which is a goal for computational psychiatry.

Ser-261 phospho-regulation is involved in pS256 and pS269-mediated aquaporin-2 apical translocation

Vasopressin catalyzes aquaporin-2 phosphorylation at several serine sites in the C-terminal region. Compared with Ser-256 and Ser-269 phosphorylation, the role of Ser-261 phospho-regulation on vasopressin-regulated AQP2 apical translocation is largely unknown. In addition, recent discovery of transcytotic apical delivery of AQP2 made the concept of its intracellular trafficking even more complicated. In this study, we evaluated how intact phospho-AQP2 signals fit with the transcytosis trafficking model in Madin-Darby canine kidney cells. PS256 and pS269 signals were intracellularly detectable in wild-type AQP2 at the beginning of forskolin stimulation (1 min). These phospho-signals were detectable in basolateral membranes even after 10 min of stimulation. AQP2 stably inserted in the apical membrane increased pS269 and decreased pS261 signals. In an NDI-causing mutant P262L-AQP2, in which Ser-261 phospho-regulation is impaired, the pS256 and pS269 signals were detectable in the basolateral membranes with increased pS261 signals after forskolin stimulation. These results suggest that Ser-261 phospho-regulation is involved in pS256- and pS269-mediated AQP2 apical translocation.

Cyclin D1 expression and polysomy in lymphocyte-predominant cells of nodular lymphocyte-predominant Hodgkin lymphoma.

Cyclin D1 protein expression in lymphocytes is classically associated with mantle cell lymphoma. Although increasingly recognized in other lymphoproliferative disorders, cyclin D1 expression and CCND1 gene abnormalities have not been well studied in nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL). Using a double stain for CD20/cyclin D1, we quantified cyclin D1 expression in 10 cases of NLPHL and correlated those findings with SOX11 expression, CCND1 gene abnormalities, and clinical data. For comparison, we examined 5 cases of T cell-/histiocyte-rich large B-cell lymphoma (THRLBCL). All cases of NLPHL stained for cyclin D1 showed at least rare positivity in lymphocyte-predominant (LP) cells. In 4 cases, at least 20% of LP cells were positive for CD20/cyclin D1. Neither SOX11 expression nor CCND1 gene rearrangement was found in any of the cases, but fluorescence in situ hybridization showed a proportion of the large cells with 3 to 4 copies of nonfused IGH and CCND1 signals or 3 intact CCND1 break-apart signals. Further study with CCND1/CEP11 showed polysomy in 6 of 9 cases with cyclin D1 expression and 5 of 16 NLPHL not examined for cyclin D1. Two of 5 cases of THRLBCL showed rare positive staining for CD20/cyclin D1; 1 case showed polysomy with CCND1/CEP11. Results show that cyclin D1 may be expressed in LP cells without SOX11 expression or CCND1 translocation. Polysomy with increased copies of CCND1 may account for cyclin D1 expression in some cases. Cyclin D1 expression is not useful for distinguishing NLPHL from THRLBCL and has no apparent clinical significance in NLPHL.