Vulval Microglandular Adenosis-like Neoplasm With Chondromyxoid Stroma: Report of a Unique Case.

Microglandular adenosis of the breast is an uncommon lesion which can mimic tubular carcinoma. It is composed of small round tubules lined by a single layer of flat or cuboidal epithelial cells, often with eosinophilic colloid-like material within the lumen. The absence of a myoepithelial cell layer and preservation of basement membrane around the tubules are characteristic features. We report the first case of a vulval "neoplasm" in a 60-yr-old woman showing features identical to microglandular adenosis of the breast except for the presence of an unusual chondromyxoid stroma. This case reinforces the analogy between vulval lesions derived from anogenital mammary-like glands and breast lesions and also adds a new entity to the list of lesions arising from these glands.

The Tamil Version of the Three-Factor Eating Questionnaire-R18V2: Reliability and Factor Structure Among Persons with Schizophrenia.

Background: Persons with schizophrenia (PwS) are vulnerable to developing disordered eating behaviors. However, standardized tools to assess disordered eating patterns are unavailable in the regional language, Tamil. Different versions of the Three-Factor Eating Questionnaire (TFEQ) have been used to measure disordered eating patterns among PwS worldwide. This study aimed to assess the factor structure and reliability of the Tamil version of TFEQ-R18V2 among Tamil-speaking PwS. Methods: Over three months, 135 PwS, aged 18-65 years, who attended the outpatient department of a tertiary mental health service provider in Chennai, completed the Tamil version of TFEQ-R18V2. Thirty PwS completed the tool after two weeks to assess its test-retest reliability. The factor structure of the tool was explored using principal component analysis. Results: The sample included 75 (55.6%) males and 60 (44.4%) females with a mean (±SD) age of 40.1 (±9.8) years and a mean duration of illness of 11.99 (± 8.72) years. Internal consistency and test-retest reliability of the Tamil version were 0.84 and 0.532, respectively. A five-factor structure emerged from the factor analysis, with 65.67% of the variance. Conclusion: The Tamil version of TFEQ-R18V2 emerged as a reliable tool to assess disordered eating patterns among Tamil-speaking PwS.

Single-cell multiomic dissection of response and resistance to chimeric antigen receptor T cells against BCMA in relapsed multiple myeloma.

Markers that predict response and resistance to chimeric antigen receptor (CAR) T cells in relapsed/refractory multiple myeloma are currently missing. We subjected mononuclear cells isolated from peripheral blood and bone marrow before and after the application of approved B cell maturation antigen-directed CAR T cells to single-cell multiomic analyses to identify markers associated with resistance and early relapse. Differences between responders and nonresponders were identified at the time of leukapheresis. Nonresponders showed an immunosuppressive microenvironment characterized by increased numbers of monocytes expressing the immune checkpoint molecule CD39 and suppressed CD8+ T cell and natural killer cell function. Analysis of CAR T cells showed cytotoxic and exhausted phenotypes in hyperexpanded clones compared to low/intermediate expanded clones. We identified potential immunotherapy targets on CAR T cells, like PD1, to improve their functionality and durability. Our work provides evidence that an immunosuppressive microenvironment causes resistance to CAR T cell therapies in multiple myeloma.

The CD47-binding peptide of thrombospondin-1 induces defenestration of liver sinusoidal endothelial cells.

BACKGROUND & AIMS: A fenestrated phenotype is characteristic of liver sinusoidal endothelial cells (LSECs), but liver sinusoids become defenestrated during fibrosis and other liver diseases. Thrombospondin-1 (TSP1) is a matrix glycoprotein with pro-fibrotic effects, and the CD47-binding fragment of TSP1 also has anti-angiogenic effects in endothelial cells. We hypothesized that the CD47-binding fragment of TSP1 could induce defenestration in LSECs through the Rho-Rho kinase (ROCK)-myosin pathway. METHODS: Freshly isolated rat LSECs were treated with TSP1 or CD47-binding peptides of TSP1. LSEC fenestration was assessed with scanning electron microscopy, and myosin phosphorylation was assessed with immuno-fluorescence. RESULTS: Treating LSECs with TSP1 caused a dose-dependent loss of fenestrae, and this effect could not be blocked by SB-431542, the TGF-ß1 receptor inhibitor. A CD47-binding fragment of TSP1, p4N1, was able to induce defenestration, and a CD47-blocking antibody, B6H12, was able to suppress p4N1-induced defenestration. The p4N1 fragment also caused contraction of fenestra size, correlated with an increase in myosin activation. Pretreatment with Y-237642 (a ROCK inhibitor) prevented p4N1-induced myosin activation and fenestrae decrease. Simvastatin has also been shown to antagonize Rho-ROCK signalling, and we found that simvastatin pretreatment protected LSECs from p4N1-induced myosin activation and defenestration. CONCLUSIONS: We conclude that CD47 signals through the Rho-ROCK-myosin pathway to induce defenestration in LSECs. In addition, our results show that simvastatin and Y-237642 have a beneficial impact on fenestration in vitro, providing an additional explanation for the efficacy of these compounds for regression of liver fibrosis.

The Tamil version of Eating Attitudes Test-26: Reliability and factor structure among persons with schizophrenia.

Background: Disordered eating behaviors (DEBs) among persons with schizophrenia (PwS) have been reported widely in the literature, with very few studies in India. Robust tools to assess DEB are needed in the vernacular language to capture symptoms of disordered eating accurately. No such tools are available in the Tamil language. Globally, the Eating Attitudes Test (EAT-26) is widely used to assess DEB among PwS. Aim: This study aimed to translate and study the factor structure and reliability of EAT-26 among Tamil-speaking PwS. Materials and Methods: EAT-26 was translated into Tamil following the Oxford linguistic validation process. Experts evaluated its face validity and content validity. One hundred and fifty PwS, aged between 18 and 65 years, who attended the outpatient department of a psychiatric facility, and consented to participate, completed the Tamil version of EAT-26. Test-retest reliability of EAT-26 was assessed by readministering the tool to 30 PwS after two weeks. Data were analyzed using Stata 16.1. Internal consistency and test-retest reliability were computed using Cronbach's alpha and intraclass coefficients, respectively. The factor structure of EAT-26 was explored using principal component analysis (PCA). Spearman's rho was calculated to understand the correlation between the factors. Results: EAT-26 had an internal consistency of 0.71 and test-retest reliability of 0.896. Factor analysis revealed nine latent factors consisting of 21 of the original 26 items on EAT-26. These 21 items could explain a variance of 63.63%. Conclusions: The Tamil version of the EAT-26 is a reliable tool to assess DEB among Tamil-speaking PwS. It can be used to screen PwS for eating disorder risk.

Assessing the effectiveness and cost-effectiveness of a solution-focused resource-orientated approach (DIALOG+) to improving the quality of life for people with psychosis in India and Pakistan-a cluster RCT.

BACKGROUND: Severe mental illness (SMI) presents a major challenge worldwide, affecting approximately 5-8% of the world's population. It causes significant distress to affected people, families and wider communities, generating high costs through loss of productivity and ongoing healthcare use. Over 75% of patients with psychosis receive inadequate care and experience a negative financial impact and reduced quality of life (QoL). It is therefore a priority to reduce the treatment gap by providing low-cost, effective interventions for people with psychosis. Our research project, PIECEs, is designed to explore, adapt and test a low-cost, approach (DIALOG+) that makes use of existing resources to improve community-based care for patients with psychosis. The research will be conducted in two urban sites: Karachi, Pakistan and Chennai, India. DIALOG+ is a novel, technology-assisted and resource-oriented intervention, based on QoL research, concepts of patient-centred communication, IT developments and solution-focused therapy. However, the approach has not been rigorously tested within India and Pakistan. Our randomised controlled trial (RCT) aims to test the effectiveness and cost-effectiveness of DIALOG+ in improving the QoL and clinical outcomes for individuals with long-term psychosis being treated in the community in India and Pakistan. METHODS: To assess the acceptability, feasibility, and cost effectiveness of DIALOG+, we will conduct a cluster RCT with 210 patients and 14 clinicians in each country. The intervention will be used during a routine interaction between a clinician and a patient. It consists of a patient-centred assessment (the DIALOG scale) whereby the clinician invites the patient to rate their satisfaction with different life domains and treatment aspects, which forms the active control group. The intervention group will follow this up with a four-step solution-focused approach to identify the patient's resources and develop solutions to deal with the patient's concerns (DIALOG+). DISCUSSION: If shown to be effective DIALOG+ has the potential to improve community-based care and the QoL for millions of people within India and Pakistan who experience psychosis. TRIAL REGISTRATION: The trial was registered prospectively on the ISRCTN Registry: ISRCTN13022816 on 9 February 2022.

Plasmin triggers a switch-like decrease in thrombospondin-dependent activation of TGF-ß1.

Transforming growth factor-ß1 (TGF-ß1) is a potent regulator of extracellular matrix production, wound healing, differentiation, and immune response, and is implicated in the progression of fibrotic diseases and cancer. Extracellular activation of TGF-ß1 from its latent form provides spatiotemporal control over TGF-ß1 signaling, but the current understanding of TGF-ß1 activation does not emphasize cross talk between activators. Plasmin (PLS) and thrombospondin-1 (TSP1) have been studied individually as activators of TGF-ß1, and in this work we used a systems-level approach with mathematical modeling and in vitro experiments to study the interplay between PLS and TSP1 in TGF-ß1 activation. Simulations and steady-state analysis predicted a switch-like bistable transition between two levels of active TGF-ß1, with an inverse correlation between PLS and TSP1. In particular, the model predicted that increasing PLS breaks a TSP1-TGF-ß1 positive feedback loop and causes an unexpected net decrease in TGF-ß1 activation. To test these predictions in vitro, we treated rat hepatocytes and hepatic stellate cells with PLS, which caused proteolytic cleavage of TSP1 and decreased activation of TGF-ß1. The TGF-ß1 activation levels showed a cooperative dose response, and a test of hysteresis in the cocultured cells validated that TGF-ß1 activation is bistable. We conclude that switch-like behavior arises from natural competition between two distinct modes of TGF-ß1 activation: a TSP1-mediated mode of high activation and a PLS-mediated mode of low activation. This switch suggests an explanation for the unexpected effects of the plasminogen activation system on TGF-ß1 in fibrotic diseases in vivo, as well as novel prognostic and therapeutic approaches for diseases with TGF-ß dysregulation.

Service providers' perceptions of the strengths and prospective improvements in UK eating disorder services: findings from a Royal College survey.

AIMS AND METHODS: This study aims to explore perceptions of eating disorder service strengths and to develop a clearer picture of improvements clinicians would like to see occur in the services they lead. A survey designed by the Royal College of Psychiatrists' Section of Eating Disorders was completed by 83 lead clinicians in both public and private sector services in the UK and Eire. Content analysis was performed, and common themes were identified. RESULTS: Five main strengths of a service were identified as follows: quality of treatment (n = 36), staff skills (n = 21), continuity of care (n = 15), family involvement (n = 12) and accessibility and availability (n = 11). These themes also arose when clinicians evaluated areas they wished to develop and improve. CONCLUSIONS: Service providers' views were congruent with each other, NICE guidelines and quality standards as proposed by the Royal College. Although clinicians feel that their service fulfils many practice guidelines, there remains areas in which adherence is felt to be lacking.

Acceptability of Peer Support for People With Schizophrenia in Chennai, India: A Cross Sectional Study Amongst People With Lived Experience, Caregivers, and Mental Health Professionals.

Introduction: Establishing structured peer support in mental health, particularly for people with schizophrenia, as a psychosocial intervention is early in low and middle-income countries like India. Before implementing and understanding the effectiveness of peer support service and which mode of peer support delivery will be suitable for our culture, our study aimed to understand if peer support would be accepted by the different participants like persons with schizophrenia, caregivers and mental health professionals in a tertiary care center in Chennai, India. Methods: The study was conducted at the outpatient department (OPD) of a tertiary psychiatric care facility in Chennai, India. A cross-sectional study method was used. Consecutive persons diagnosed with schizophrenia and caregivers of persons with schizophrenia, who attended the outpatient department, and mental health professionals within and outside the facility who met the inclusion and exclusion criteria participated in the study. A structured questionnaire purposefully developed for the study was administered to the different study participants. Descriptive statistics were used to analyze the data. Categorical variables were expressed as frequency and percentages, while the continuous variables were expressed as mean and standard deviation. Results: A total of 155 participants (52 persons with schizophrenia, 50 caregivers and 53 mental health professionals) completed the survey. The majority of the participants (90.4% of persons with schizophrenia, 86% caregivers and all mental health professionals) welcomed peer support interventions. The participants wanted peers to help persons with schizophrenia achieve personal goals to enhance their mental health and day to day living with an emphasis on independent living and interpersonal and social relationships and help them achieve medication and treatment-related goals toward recovery. Understanding the role of a peer support volunteer and transitioning from a "person with schizophrenia" to a "peer support volunteer" by persons with schizophrenia was thought most challenging. Conclusion: The results highlight the potential acceptability of peer support across several stakeholders in the care of schizophrenia in a low and middle-income country context. The results may guide the implementation of a peer support volunteer programme as an essential mechanism of delivering psychosocial interventions for persons with schizophrenia.

Implementing psychosocial interventions within low and middle-income countries to improve community-based care for people with psychosis-A situation analysis.

Background: Globally, a treatment gap exists for individuals with severe mental illness, with 75% of people with psychosis failing to receive appropriate care. This is most pronounced in low and middle-income countries, where there are neither the financial nor human resources to provide high-quality community-based care. Low-cost, evidence-based interventions are urgently needed to address this treatment gap. Aim: To conduct a situation analysis to (i) describe the provision of psychosocial interventions within the context of existing care in two LMICs-India and Pakistan, and (ii) understand the barriers and facilitators of delivering a new psychosocial intervention. Method: A situation analysis including a quantitative survey and individual interviews with clinicians, patients and caregivers was conducted. Quantitative survey data was collected from staff members at 11 sites (private and government run hospitals) to assess organizational readiness to implement a new psychosocial intervention. To obtain in-depth information, 24 stakeholders including clinicians and service managers were interviewed about the typical care they provide and/or receive, and their experience of either accessing or delivering psychosocial interventions. This was triangulated by six interviews with carer and patient representatives. Results and discussion: The results highlight the positive views toward psychosocial interventions within routine care and the enthusiasm for multidisciplinary working. However, barriers to implementation such as clinician time, individual attitudes toward psychosocial interventions and organizational concerns including the lack of space within the facility were highlighted. Such barriers need to be taken into consideration when designing how best to implement and sustain new psychosocial interventions for the community treatment of psychosis within LMICs.

Steady states and dynamics of urokinase-mediated plasmin activation in silico and in vitro.

Plasmin (PLS) and urokinase-type plasminogen activator (UPA) are ubiquitous proteases that regulate the extracellular environment. Although they are secreted in inactive forms, they can activate each other through proteolytic cleavage. This mutual interplay creates the potential for complex dynamics, which we investigated using mathematical modeling and in vitro experiments. We constructed ordinary differential equations to model the conversion of precursor plasminogen into active PLS, and precursor urokinase (scUPA) into active urokinase (tcUPA). Although neither PLS nor UPA exhibits allosteric cooperativity, modeling showed that cooperativity occurred at the system level because of substrate competition. Computational simulations and bifurcation analysis predicted that the system would be bistable over a range of parameters for cooperativity and positive feedback. Cell-free experiments with recombinant proteins tested key predictions of the model. PLS activation in response to scUPA stimulus was found to be cooperative in vitro. Finally, bistability was demonstrated in vitro by the presence of two significantly different steady-state levels of PLS activation for the same levels of stimulus. We conclude that ultrasensitive, bistable activation of UPA-PLS is possible in the presence of substrate competition. An ultrasensitive threshold for activation of PLS and UPA would have ramifications for normal and disease processes, including angiogenesis, metastasis, wound healing, and fibrosis.

Intra-vessel heterogeneity establishes enhanced sites of macromolecular leakage downstream of laminin α5.

Endothelial cells display heterogeneous properties based on location and function. How this heterogeneity influences endothelial barrier stability both between and within vessel subtypes is unexplored. In this study, we find that endothelial cells exhibit heterogeneous barrier properties on inter-organ and intra-vessel levels. Using intravital microscopy and sequential stimulation of the ear dermis with vascular endothelial growth factor-A (VEGFA) and/or histamine, we observe distinct, reappearing sites, common for both agonists, where leakage preferentially takes place. Through repetitive stimulation of the diaphragm and trachea, we find inter-organ conservation of such predetermined leakage sites. Qualitatively, predetermined sites display distinct leakage properties and enhanced barrier breakdown compared to less susceptible regions. Mechanistically, laminin α5 is reduced at predetermined sites, which is linked to reduced junctional vascular endothelial (VE)-cadherin and enhanced VEGFA-induced VE-cadherin phosphorylation. These data highlight functional intra-vessel heterogeneity that defines predetermined sites with distinct leakage properties and that may disproportionately impact pathological vascular leakage.

Positive Feedback Defines the Timing, Magnitude, and Robustness of Angiogenesis.

Angiogenesis is driven by the coordinated collective branching of specialized leading "tip" and trailing "stalk" endothelial cells (ECs). While Notch-regulated negative feedback suppresses excessive tip selection, roles for positive feedback in EC identity decisions remain unexplored. Here, by integrating computational modeling with in vivo experimentation, we reveal that positive feedback critically modulates the magnitude, timing, and robustness of angiogenic responses. In silico modeling predicts that positive-feedback-mediated amplification of VEGF signaling generates an ultrasensitive bistable switch that underpins quick and robust tip-stalk decisions. In agreement, we define a positive-feedback loop exhibiting these properties in vivo, whereby Vegf-induced expression of the atypical tetraspanin, tm4sf18, amplifies Vegf signaling to dictate the speed and robustness of EC selection for angiogenesis. Consequently, tm4sf18 mutant zebrafish select fewer motile ECs and exhibit stunted hypocellular vessels with unstable tip identity that is severely perturbed by even subtle Vegfr attenuation. Hence, positive feedback spatiotemporally shapes the angiogenic switch to ultimately modulate vascular network topology.

Improved clonality assessment in germinal centre/post-germinal centre non-Hodgkin's lymphomas with high rates of somatic hypermutation.

BACKGROUND: PCR detects clonal rearrangements of the Ig gene in lymphoproliferative disorders. False negativity occurs in germinal centre/post-germinal centre lymphomas (GC/PGCLs) as they display a high rate of somatic hypermutation (SHM), which causes primer mismatching when detecting Ig rearrangements by PCR. AIMS: To investigate the degree of SHM in a group of GC/PGCLs and assess the rate of false negativity when using BIOMED-2 PCR when compared with previously published strategies. METHODS: DNA was isolated from snap-frozen tissue from 49 patients with GC/PGCL (23 diffuse large B cell lymphomas (DLBCLs), 26 follicular lymphomas (FLs)) and PCR-amplified for complete (VDJH), incomplete (DJH) and Ig kappa/lambda rearrangements using the BIOMED-2 protocols, and compared with previously published methods using consensus primers. Germinal centre phenotype was defined by immunohistochemistry based on CD10, Bcl-6 and MUM-1. RESULTS: Clonality detection by amplifying Ig rearrangements using BIOMED-2 family-specific primers was considerably higher than that found using consensus primers (74% DLBCL and 96% FL vs 69% DLBCL and 73% FL). Addition of BIOMED-2 DJH rearrangements increased detection of clonality by 22% in DLBCL. SHM was present in VDJH rearrangements from all patients with DLBCL (median (range) 5.7% (2.5-13.5)) and FL (median (range) 5.3% (2.3-11.9)) with a clonal rearrangement. CONCLUSIONS: Use of BIOMED-2 primers has significantly reduced the false negative rate associated with GC/PGCL when compared with consensus primers, and the inclusion of DJH rearrangements represents a potential complementary target for clonality assessment, as SHM is thought not to occur in these types of rearrangements.

Computational analysis reveals the coupling between bistability and the sign of a feedback loop in a TGF-ß1 activation model.

BACKGROUND: Bistable behaviors are prevalent in cell signaling and can be modeled by ordinary differential equations (ODEs) with kinetic parameters. A bistable switch has recently been found to regulate the activation of transforming growth factor-ß1 (TGF-ß1) in the context of liver fibrosis, and an ordinary differential equation (ODE) model was published showing that the net activation of TGF-ß1 depends on the balance between two antagonistic sub-pathways. RESULTS: Through modeling the effects of perturbations that affect both sub-pathways, we revealed that bistability is coupled with the signs of feedback loops in the model. We extended the model to include calcium and Krüppel-like factor 2 (KLF2), both regulators of Thrombospondin-1 (TSP1) and Plasmin (PLS). Increased levels of extracellular calcium, which alters the TSP1-PLS balance, would cause high levels of TGF-ß1, resembling a fibrotic state. KLF2, which suppresses production of TSP1 and plasminogen activator inhibitor-1 (PAI1), would eradicate bistability and preclude the fibrotic steady-state. Finally, the loop PLS - TGF-ß1 - PAI1 had previously been reported as negative feedback, but the model suggested a stronger indirect effect of PLS down-regulating PAI1 to produce positive (double-negative) feedback in a fibrotic state. Further simulations showed that activation of KLF2 was able to restore negative feedback in the PLS - TGF-ß1 - PAI1 loop. CONCLUSIONS: Using the TGF-ß1 activation model as a case study, we showed that external factors such as calcium or KLF2 can induce or eradicate bistability, accompanied by a switch in the sign of a feedback loop (PLS - TGF-ß1 - PAI1) in the model. The coupling between bistability and positive/negative feedback suggests an alternative way of characterizing a dynamical system and its biological implications.

Time to Decide? Dynamical Analysis Predicts Partial Tip/Stalk Patterning States Arise during Angiogenesis.

Angiogenesis is a highly dynamic morphogenesis process; however, surprisingly little is known about the timing of the different molecular processes involved. Although the role of the VEGF-notch-DLL4 signaling pathway has been established as essential for tip/stalk cell competition during sprouting, the speed and dynamic properties of the underlying process at the individual cell level has not been fully elucidated. In this study, using mathematical modeling we investigate how specific, biologically meaningful, local conditions around and within an individual cell can influence their unique tip/stalk phenotype switching kinetics. To this end we constructed an ordinary differential equation model of VEGF-notch-DLL4 signaling in a system of two, coupled endothelial cells (EC). Our studies reveal that at any given point in an angiogenic vessel the time it takes a cell to decide to take on a tip or stalk phenotype may be drastically different, and this asynchrony of tip/stalk cell decisions along vessels itself acts to speed up later competitions. We unexpectedly uncover intermediate "partial" yet stable states lying between the tip and stalk cell fates, and identify that internal cellular factors, such as NAD-dependent deacetylase sirtuin-1 (Sirt1) and Lunatic fringe 1 (Lfng1), can specifically determine the length of time a cell spends in these newly identified partial tip/stalk states. Importantly, the model predicts that these partial EC states can arise during normal angiogenesis, in particular during cell rearrangement in sprouts, providing a novel two-stage mechanism for rapid adaptive behavior to the cells highly dynamic environment. Overall, this study demonstrates that different factors (both internal and external to EC) can be used to modulate the speed of tip/stalk decisions, opening up new opportunities and challenges for future biological experiments and therapeutic targeting to manipulate vascular network topology, and our basic understanding of developmental/pathological angiogenesis.

Protrusive waves guide 3D cell migration along nanofibers.

In vivo, cells migrate on complex three-dimensional (3D) fibrous matrices, which has made investigation of the key molecular and physical mechanisms that drive cell migration difficult. Using reductionist approaches based on 3D electrospun fibers, we report for various cell types that single-cell migration along fibronectin-coated nanofibers is associated with lateral actin-based waves. These cyclical waves have a fin-like shape and propagate up to several hundred micrometers from the cell body, extending the leading edge and promoting highly persistent directional movement. Cells generate these waves through balanced activation of the Rac1/N-WASP/Arp2/3 and Rho/formins pathways. The waves originate from one major adhesion site at leading end of the cell body, which is linked through actomyosin contractility to another site at the back of the cell, allowing force generation, matrix deformation and cell translocation. By combining experimental and modeling data, we demonstrate that cell migration in a fibrous environment requires the formation and propagation of dynamic, actin based fin-like protrusions.