The First Transcriptomic Atlas of the Adult Lacrimal Gland Reveals Epithelial Complexity and Identifies Novel Progenitor Cells in Mice.

The lacrimal gland (LG) secretes aqueous tears. Previous studies have provided insights into the cell lineage relationships during tissue morphogenesis. However, little is known about the cell types composing the adult LG and their progenitors. Using scRNAseq, we established the first comprehensive cell atlas of the adult mouse LG to investigate the cell hierarchy, its secretory repertoire, and the sex differences. Our analysis uncovered the complexity of the stromal landscape. Epithelium subclustering revealed myoepithelial cells, acinar subsets, and two novel acinar subpopulations: Tfrchi and Car6hi cells. The ductal compartment contained Wfdc2+ multilayered ducts and an Ltf+ cluster formed by luminal and intercalated duct cells. Kit+ progenitors were identified as: Krt14+ basal ductal cells, Aldh1a1+ cells of Ltf+ ducts, and Sox10+ cells of the Car6hi acinar and Ltf+ epithelial clusters. Lineage tracing experiments revealed that the Sox10+ adult populations contribute to the myoepithelial, acinar, and ductal lineages. Using scRNAseq data, we found that the postnatally developing LG epithelium harbored key features of putative adult progenitors. Finally, we showed that acinar cells produce most of the sex-biased lipocalins and secretoglobins detected in mouse tears. Our study provides a wealth of new data on LG maintenance and identifies the cellular origin of sex-biased tear components.

Psychotherapist advice, suggestions, recommendations: A research review.

Psychotherapists provide at least some advice, suggestions, and recommendations (ASR) in most treatment approaches. We define ASR, offer clinical examples, and review the research evidence for the immediate in-session, immediate delayed, and intermediate effects of ASR, as well as for the moderators of these effects in individual psychotherapy. In seven studies with 327 clients and 131 therapists, we found evidence of neutral immediate in-session outcomes (e.g., client experiencing levels), neutral immediate delayed outcomes (e.g., client-rated helpfulness during postsession videotape reviews), and positive intermediate outcomes (e.g., implementation of recommendations as rated in subsequent sessions) for ASR. These differences may be related to methodological variations in studies in addition to the different timing of the outcome measurement. In terms of moderators, there is some evidence that the working alliance, client collaboration prior to the ASR, content of the ASR, and therapist and client attachment styles moderate the effects of immediate in-session outcomes, and that type, difficulty, and therapist influence moderate the effects of intermediate outcomes of ASR. We conclude with research limitations, training implications, and therapeutic practices related to ASR. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Patient perspectives on working with preferences in psychotherapy: A consensual qualitative research study.

OBJECTIVE: Assessing and accommodating patient preferences is integral to evidence-based practice. This qualitative study sought to explore patient perspectives and experiences of preference work in psychotherapy. METHODS: Participants were 13 UK-based patients who had completed up to 24 sessions of a collaborative-integrative psychotherapy. Ten participants identified as female and three as male. Interviews were conducted at endpoint and analyzed using a team-based, consensual qualitative research approach. RESULTS: Three superordinate domains were developed: Preferences Themselves, Process of Working with Preferences in Psychotherapy, and Effect of Preference Work (or its Absence). Patients typically wanted leadership, challenge, and input from their psychotherapist, and an affirming style. Patients attributed the origin of their preferences to personal history, characteristics, or circumstances; the present psychotherapy; or past episodes of psychotherapy. Some preferences changed over time. Preference work was described as having positive effects on the therapeutic relationship and patients' intrapersonal worlds; however, variantly, non-accommodation of preferences was also experienced as beneficial. CONCLUSION: Our findings provide in-depth answers to a range of novel questions on preference work-potential mechanisms by which preference work impacts outcomes, factors that may facilitate preference work, and origins of preferences-as well as nuancing previously-established quantitative findings. Implications for clinical training and practice are discussed.

Client perspectives on psychotherapy failure.

This study qualitatively examined client's definition and experiences of failed psychotherapy.Thirteen clients were interviewed by phone regarding their experience of failed psychotherapy. Data were analyzed using consensual qualitative research (CQR).Participants defined failed psychotherapy as negatively affecting clients, involving problems in the psychotherapy relationship, and not meeting clients' goals. When describing specific experiences of failed psychotherapy, participants gradually recognized the failure themselves, but the recognition was sometimes facilitated by others. Pre-termination, the failed psychotherapy yielded negative effects (worsened symptoms/functioning, deteriorating relationship, not addressing clients' concerns). Participants perceived therapists' contributions as involving action (insensitive/inappropriate responses to participants' concerns about psychotherapy) and inaction (not managing psychotherapy effectively). They perceived their own contributions as their difficulty voicing their concerns or asserting themselves. Post-termination effects were negative cognitively/affectively (heightened distress), behaviorally (disinterest in seeking mental health services), and interpersonally (relationship difficulties in later psychotherapy); the failed psychotherapy also helped participants pursue their needs in psychotherapy.Failed psychotherapy consisted of problems in the relationship and the treatment not meeting client's goals. Such psychotherapy worsened clients' functioning, further damaged an already tenuous psychotherapy relationship, and both therapists and clients contributed to the failure. After termination, failed psychotherapy yielded cognitive/affective, behavioral, and interpersonal effects.

Long-term functional regeneration of radiation-damaged salivary glands through delivery of a neurogenic hydrogel.

Salivary gland acinar cells are severely depleted after radiotherapy for head and neck cancer, leading to loss of saliva and extensive oro-digestive complications. With no regenerative therapies available, organ dysfunction is irreversible. Here, using the adult murine system, we demonstrate that radiation-damaged salivary glands can be functionally regenerated via sustained delivery of the neurogenic muscarinic receptor agonist cevimeline. We show that endogenous gland repair coincides with increased nerve activity and acinar cell division that is limited to the first week after radiation, with extensive acinar cell degeneration, dysfunction, and cholinergic denervation occurring thereafter. However, we found that mimicking cholinergic muscarinic input via sustained local delivery of a cevimeline-alginate hydrogel was sufficient to regenerate innervated acini and retain physiological saliva secretion at nonirradiated levels over the long term (>3 months). Thus, we reveal a previously unknown regenerative approach for restoring epithelial organ structure and function that has extensive implications for human patients.

Neuronal-epithelial cross-talk drives acinar specification via NRG1-ERBB3-mTORC2 signaling.

Acinar cells are the principal secretory units of multiple exocrine organs. A single-cell, layered, lumenized acinus forms from a large cohort of epithelial progenitors that must initiate and coordinate three cellular programs of acinar specification, namely, lineage progression, secretion, and polarization. Despite this well-known outcome, the mechanism(s) that regulate these complex programs are unknown. Here, we demonstrate that neuronal-epithelial cross-talk drives acinar specification through neuregulin (NRG1)-ERBB3-mTORC2 signaling. Using single-cell and global RNA sequencing of developing murine salivary glands, we identified NRG1-ERBB3 to precisely overlap with acinar specification during gland development. Genetic deletion of Erbb3 prevented cell lineage progression and the establishment of lumenized, secretory acini. Conversely, NRG1 treatment of isolated epithelia was sufficient to recapitulate the development of secretory acini. Mechanistically, we found that NRG1-ERBB3 regulates each developmental program through an mTORC2 signaling pathway. Thus, we reveal that a neuronal-epithelial (NRG1/ERBB3/mTORC2) mechanism orchestrates the creation of functional acini.

A mechanism of gene evolution generating mucin function.

How novel gene functions evolve is a fundamental question in biology. Mucin proteins, a functionally but not evolutionarily defined group of proteins, allow the study of convergent evolution of gene function. By analyzing the genomic variation of mucins across a wide range of mammalian genomes, we propose that exonic repeats and their copy number variation contribute substantially to the de novo evolution of new gene functions. By integrating bioinformatic, phylogenetic, proteomic, and immunohistochemical approaches, we identified 15 undescribed instances of evolutionary convergence, where novel mucins originated by gaining densely O-glycosylated exonic repeat domains. Our results suggest that secreted proteins rich in proline are natural precursors for acquiring mucin function. Our findings have broad implications for understanding the role of exonic repeats in the parallel evolution of new gene functions, especially those involving protein glycosylation.

A synthetic tear protein resolves dry eye through promoting corneal nerve regeneration.

Corneal architecture is essential for vision and is greatly perturbed by the absence of tears due to the highly prevalent disorder dry eye. With no regenerative therapies available, pathological alterations of the ocular surface in response to dryness, including persistent epithelial defects and poor wound healing, result in lifelong morbidity. Here, using a mouse model of aqueous-deficient dry eye, we reveal that topical application of the synthetic tear protein Lacripep reverses the pathological outcomes of dry eye through restoring the extensive network of corneal nerves that are essential for tear secretion, barrier function, epithelial homeostasis, and wound healing. Intriguingly, the restorative effects of Lacripep occur despite extensive immune cell infiltration, suggesting tissue reinnervation and regeneration can be achieved under chronic inflammatory conditions. In summary, our data highlight Lacripep as a first-in-class regenerative therapy for returning the cornea to a near homeostatic state in individuals who suffer from dry eye.

Aldehyde dehydrogenase 3A1 deficiency leads to mitochondrial dysfunction and impacts salivary gland stem cell phenotype.

Adult salivary stem/progenitor cells (SSPC) have an intrinsic property to self-renew in order to maintain tissue architecture and homeostasis. Adult salivary glands have been documented to harbor SSPC, which have been shown to play a vital role in the regeneration of the glandular structures postradiation damage. We have previously demonstrated that activation of aldehyde dehydrogenase 3A1 (ALDH3A1) after radiation reduced aldehyde accumulation in SSPC, leading to less apoptosis and improved salivary function. We subsequently found that sustained pharmacological ALDH3A1 activation is critical to enhance regeneration of murine submandibular gland after radiation damage. Further investigation shows that ALDH3A1 function is crucial for SSPC self-renewal and survival even in the absence of radiation stress. Salivary glands from Aldh3a1 -/- mice have fewer acinar structures than wildtype mice. ALDH3A1 deletion or pharmacological inhibition in SSPC leads to a decrease in mitochondrial DNA copy number, lower expression of mitochondrial specific genes and proteins, structural abnormalities, lower membrane potential, and reduced cellular respiration. Loss or inhibition of ALDH3A1 also elevates ROS levels, depletes glutathione pool, and accumulates ALDH3A1 substrate 4-hydroxynonenal (4-HNE, a lipid peroxidation product), leading to decreased survival of murine SSPC that can be rescued by treatment with 4-HNE specific carbonyl scavengers. Our data indicate that ALDH3A1 activity protects mitochondrial function and is important for the regeneration activity of SSPC. This knowledge will help to guide our translational strategy of applying ALDH3A1 activators in the clinic to prevent radiation-related hyposalivation in head and neck cancer patients.

Exocrine gland structure-function relationships.

Fluid secretion by exocrine glandular organs is essential to the survival of mammals. Each glandular unit within the body is uniquely organized to carry out its own specific functions, with failure to establish these specialized structures resulting in impaired organ function. Here, we review glandular organs in terms of shared and divergent architecture. We first describe the structural organization of the diverse glandular secretory units (the end-pieces) and their fluid transporting systems (the ducts) within the mammalian system, focusing on how tissue architecture corresponds to functional output. We then highlight how defects in development of end-piece and ductal architecture impacts secretory function. Finally, we discuss how knowledge of exocrine gland structure-function relationships can be applied to the development of new diagnostics, regenerative approaches and tissue regeneration.

Septum submucosal glands exhibit aberrant morphology and reduced mucin production in chronic rhinosinusitis.

BACKGROUND: Chronic rhinosinusitis (CRS) is characterized by significant accumulation and thickening of mucus in the sinonasal cavities. One contributor of aberrant mucus production and impaired mucociliary clearance (MCC) is altered function of the sinonasal submucosal glands (SMGs), yet contributions of SMGs to upper airway disease initiation and progression remain unknown. The objective of this study was to characterize the morphology and secretory cell identities of the nasal septum SMGs in both healthy and CRS adults. METHODS: Biopsies from adult participants with CRS without nasal polyps (CRSsNP, n = 4), CRS with nasal polyps (CRSwNP, n = 8), and non-CRS controls (n = 14) were collected from the posterior septum. Glandular morphology and mucus markers were investigated using histological techniques and high-resolution confocal microscopy. RESULTS: Analysis revealed a significant decrease in gland density in the posterior septum of CRSsNP (28% ± 6.15%) and CRSwNP (23% ± 3.09%) compared to control participants (53% ± 1.59%, p < 0.0001). Further analysis of the CRS SMG secretory function revealed an overall decrease in Mucin 5B+ gland mucus being produced. Dilated and cystic ductal structures filled with inspissated mucus were also common to CRS glands. CONCLUSION: Here, we describe a significant alteration in SMG structure and function in the adult CRS posterior septum suggesting reduced gland contribution to MCC. The SMGs of both the nose and sinuses may represent targets for future therapeutic approaches.

Twenty-Five-Year Changes in Office and Ambulatory Blood Pressure: Results From the Coronary Artery Risk Development in Young Adults (CARDIA) Study.

BACKGROUND: Blood pressure (BP) measured in the office setting increases from early through later adulthood. However, it is unknown to what extent out-of-office BP derived via ambulatory BP monitoring (ABPM) increases over time, and which participant characteristics and risk factors might contribute to these increases. METHODS: We assessed 25-year change in office- and ABPM-derived BP across sex, race, diabetes mellitus (DM), and body mass index (BMI) subgroups in the Coronary Artery Risk Development in Young Adults study using multivariable-adjusted linear mixed effects models. RESULTS: We included 288 participants who underwent ABPM at the Year 5 Exam (mean [SD] age, 25.1 [3.7]; 45.8% men) and 455 participants who underwent ABPM at the Year 30 Exam (mean [SD] age, 49.5 [3.7]; 42.0% men). Office, daytime, and nighttime systolic BP (SBP) increased 12.8 (95% confidence interval [CI], 7.6-17.9), 14.7 (95% CI, 9.7-19.8), and 16.6 (95% CI, 11.4-21.8) mm Hg, respectively, over 25 years. Office SBP increased 6.5 (95% CI, 2.3-10.6) mm Hg more among black compared with white participants. Daytime SBP increased 6.3 (95% CI, 0.2-12.4) mm Hg more among participants with a BMI ≥25 vs. <25 kg/m2. Nighttime SBP increased 4.7 (95% CI, 0.5-8.9) mm Hg more among black compared with white participants, and 17.3 (95% CI, 7.2-27.4) mm Hg more among participants with vs. without DM. CONCLUSIONS: Office- and ABPM-derived BP increased more from early through middle adulthood among black adults and participants with DM and BMI ≥25 kg/m2.

Functional Specialization of Human Salivary Glands and Origins of Proteins Intrinsic to Human Saliva.

Salivary proteins are essential for maintaining health in the oral cavity and proximal digestive tract, and they serve as potential diagnostic markers for monitoring human health and disease. However, their precise organ origins remain unclear. Through transcriptomic analysis of major adult and fetal salivary glands and integration with the saliva proteome, the blood plasma proteome, and transcriptomes of 28+ organs, we link human saliva proteins to their source, identify salivary-gland-specific genes, and uncover fetal- and adult-specific gene repertoires. Our results also provide insights into the degree of gene retention during gland maturation and suggest that functional diversity among adult gland types is driven by specific dosage combinations of hundreds of transcriptional regulators rather than by a few gland-specific factors. Finally, we demonstrate the heterogeneity of the human acinar cell lineage. Our results pave the way for future investigations into glandular biology and pathology, as well as saliva's use as a diagnostic fluid.

Alterations in corneal biomechanics underlie early stages of autoimmune-mediated dry eye disease.

Autoimmune-mediated dry eye disease is a pathological feature of multiple disorders including Sjögren's syndrome, lupus and rheumatoid arthritis that has a life-long, detrimental impact on vision and overall quality of life. Although late stage disease outcomes such as epithelial barrier dysfunction, reduced corneal innervation and chronic inflammation have been well characterized in both human patients and mouse models, there is little to no understanding of early pathological processes. Moreover, the mechanisms underlying the loss of cornea homeostasis and disease progression are unknown. Here, we utilize the autoimmune regulatory (Aire)-deficient mouse model of autoimmune-mediated dry eye disease in combination with genome wide transcriptomics, high-resolution imaging and atomic force microscopy to reveal a potential extracellular matrix (ECM)-biomechanical-based mechanism driving cellular and morphological changes at early disease onset. Early disease in the Aire-deficient mouse model is associated with a mild reduction in tear production and moderate immune cell infiltration, allowing for interrogation of cellular, molecular and biomechanical changes largely independent of chronic inflammation. Using these tools, we demonstrate for the first time that the emergence of autoimmune-mediated dry eye disease is associated with an alteration in the biomechanical properties of the cornea. We reveal a dramatic disruption of the synthesis and organization of the extracellular matrix as well as degradation of the epithelial basement membrane during early disease. Notably, we provide evidence that the nerve supply to the cornea is severely reduced at early disease stages and that this is independent of basement membrane destruction or significant immune cell infiltration. Furthermore, diseased corneas display spatial heterogeneity in mechanical, structural and compositional changes, with the limbal compartment often exhibiting the opposite response compared to the central cornea. Despite these differences, however, epithelial hyperplasia is apparent in both compartments, possibly driven by increased activation of IL-1R1 and YAP signaling pathways. Thus, we reveal novel perturbations in corneal biomechanics, matrix organization and cell behavior during the early phase of dry eye that may underlie disease development and progression, presenting new potential targets for therapeutic intervention.

Barriers to Mental Health Treatment in the Saudi National Mental Health Survey.

Objectives: To examine barriers to initiation and continuation of treatment among individuals with common mental disorders in the Saudi National Mental Health Survey (SNMHS). Methods: The SNMHS is a community-based epidemiological survey in a nationally representative household sample of respondents aged 15-65 in the Kingdom of Saudi Arabia. The World Health Organization Composite International Diagnostic Interview (CIDI) 3.0 was used. Predictors of barriers to treatment were analyzed with multivariable logistic regression. Results: Among participants with a 12-month DSM-IV/CIDI disorder (n = 711), 86.1% reported no service use. Of those (n = 597), 50.7% did not think they needed any help (categorized as "low perceived need") and 49.3% did perceive need. Of those who perceived need (n = 309), the majority (98.9%) reported attitudinal barriers to initiation. In contrast, 10.3% of those with a perceived need reported structural barriers. Respondents who were previously married or indicated below-average income were more likely to believe they needed help. Conclusions: Among people with a diagnosed mental disorder, low perceived need and attitudinal barriers are the primary barriers to mental health treatment in the KSA. The results suggest that addressing poor mental health literacy may be essential factor in reducing the unmet need for mental health treatment in the KSA.

Roadmap for the Emerging Field of Cancer Neuroscience.

Mounting evidence indicates that the nervous system plays a central role in cancer pathogenesis. In turn, cancers and cancer therapies can alter nervous system form and function. This Commentary seeks to describe the burgeoning field of "cancer neuroscience" and encourage multidisciplinary collaboration for the study of cancer-nervous system interactions.

Reservation-Urban Comparison of Suicidal Ideation/Planning and Attempts in American Indian Youth.

BACKGROUND: Our aim was to identify sex- and location-specific risk factors for suicide ideation/planning and attempts among American Indian youth. METHODS: Biennial data for 6417 American Indian high school students attending reservation and urban schools were extracted from the Montana volunteer sample Youth Risk Behavior Survey data for pooled years 2003 to 2011. Logistic regression was used to identify sex- and school location-specific risk behaviors and psychosocial factors for past 12-month ideation/planning and past 12-month attempts. RESULTS: Contrary to our hypothesis, the prevalence of ideation/planning and attempts did not significantly differ between reservation/urban location; however, risk factors associated with suicidality did. Sadness/hopelessness was associated with both outcomes for all groups. However, violent victimization was associated with both outcomes only among girls. Lack of school safety was associated with attempts but not ideation/planning among all students. There were distinct differences in risk factors associated with both outcomes among boys. CONCLUSIONS: The results indicate differences and similarities in risk behaviors and psychosocial factors associated with suicidality by sex and reservation/urban setting. Implications include screening potentially at-risk students for depression, violent victimization, substance use, and school safety and use of the findings by tribal and school programs in designing prevention and intervention programs.

Actigraphy-Based Assessment of Sleep Parameters.

Actigraphy, a method for inferring sleep/wake patterns based on movement data gathered using actigraphs, is increasingly used in population-based epidemiologic studies because of its ability to monitor activity in natural settings. Using special software, actigraphic data are analyzed to estimate a range of sleep parameters. To date, despite extensive application of actigraphs in sleep research, published literature specifically detailing the methodology for derivation of sleep parameters is lacking; such information is critical for the appropriate analysis and interpretation of actigraphy data. Reporting of sleep parameters has also been inconsistent across studies, likely reflecting the lack of consensus regarding the definition of sleep onset and offset. In addition, actigraphy data are generally underutilized, with only a fraction of the sleep parameters generated through actigraphy routinely used in current sleep research. The objectives of this paper are to review existing algorithms used to estimate sleep/wake cycles from movement data, demonstrate the rules/methods used for estimating sleep parameters, provide clear technical definitions of the parameters, and suggest potential new measures that reflect intraindividual variability. Utilizing original data collected using Motionlogger Sleep Watch (Ambulatory Monitoring Inc., Ardsley, NY), we detail the methodology and derivation of 29 nocturnal sleep parameters, including those both widely and rarely utilized in research. By improving understanding of the actigraphy process, the information provided in this paper may help: ensure appropriate use and interpretation of sleep parameters in future studies; enable the recalibration of sleep parameters to address specific goals; inform the development of new measures; and increase the breadth of sleep parameters used.

The emerging role of cranial nerves in shaping craniofacial development.

Organs and structures of the vertebrate head perform a plethora of tasks including visualization, digestion, vocalization/communication, auditory functions, and respiration in response to neuronal input. This input is primarily derived from afferent and efferent fibers of the cranial nerves (sensory and motor respectively) and efferent fibers of the cervical sympathetic trunk. Despite their essential contribution to the function and integration of processes necessary for survival, how organ innervation is established remains poorly understood. Furthermore, while it has been appreciated for some time that innervation of organs by cranial nerves is regulated in part by secreted factors and cell surface ligands expressed by those organs, whether nerves also regulate the development of facial organs is only beginning to be elucidated. This review will provide an overview of cranial nerve development in relation to the organs they innervate, and outline their known contributions to craniofacial development, thereby providing insight into how nerves may shape the organs they innervate during development. Throughout, the interaction between different cell and tissue types will be highlighted.

Deciphering Molecular and Phenotypic Changes Associated with Early Autoimmune Disease in the Aire-Deficient Mouse Model of Sjögren's Syndrome.

Sjögren's syndrome (SS) is characterized by extensive lymphocytic infiltration of the salivary and lacrimal gland (LG), resulting in acinar cell destruction and organ dysfunction. The underlying pathogenesis of SS remains largely unknown, and studies historically focus on defining late-stage disease. Here, we identify tissue programs associated with disease onset using transcriptomic and immunohistological analysis of LGs from 5- and 7-week-old mice deficient in autoimmune response element (Aire). At 5 weeks of age (wk), Aire-/- mice show minimal tissue dysfunction and destruction compared to 7 wk Aire-/-, which exhibit severe dry eye, poor tear secretion, extensive lymphocytic infiltration, reduced functional innervation, and increased vascularization. Despite this mild phenotype, 5 wk Aire-/- LGs were highly enriched for signaling pathways previously associated with SS, including interferon gamma (IFNγ), interleukin 1 beta (IL1ß), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), toll-like receptor (TLR) signaling, and interleukin-6/signal transducer and activator of transcription 3 (IL6/STAT3) signaling. Novel signaling pathways such as the semaphorin⁻plexin pathway were also noted. Intriguingly, we found an expansion of the ductal network with increasing disease. Activated STAT3, a blocker of apoptosis, was restricted to the ductal system and also increased with damage, highlighting its potential as a promoter of ductal cell survival. These data demonstrate the early activation of signaling pathways regulating inflammation, innervation, and cell survival before the onset of clinical disease indicators, suggesting their potential value as diagnostic biomarkers.