Effect of testicular degeneration on expression of sperm protein at 22 kDa in stallions.

BACKGROUND: Sperm protein at 22 kDa has been associated with fertility. OBJECTIVES: The objectives of this study were to determine (1) the localization pattern of SP22 on ejaculated and caudal epididymal equine spermatozoa and in epididymal fluid, and to (2) characterize SP22 protein and mRNA expression in testicular and epididymal tissues in response to heat-induced testicular degeneration. MATERIALS AND METHODS: Semen was collected before and after hemi-castration, as well as prior to and following insulation of the remaining testes, and tissue specimens were collected for analysis. RESULTS: Histopathology confirmed degeneration in insulated testes. Ejaculated and epididymal spermatozoa from samples collected prior to insulation of the testicles had a predominant staining pattern of SP22 over the equatorial region. However, the equatorial pattern in the pre-insulation epididymal semen samples was significantly lower than in the pre-insulation ejaculated semen samples (68 ± 3, 81 ± 2.6, respectively). Ejaculated and epididymal samples collected after insulation of the testicles showed a complete loss of staining as the predominant pattern. Western blot analysis verified the presence of SP22 on fresh ejaculated spermatozoa prior to and following heat-induced degeneration, on epididymal spermatozoa after testicular insulation, and in testicular and epididymal tissues. Heat insulation significantly reduced messenger RNA expression in the head of the epididymis and testicular tissues. Immunohistochemistry of the testicular and epididymal tissues pre-heating showed considerably weaker staining than the same tissues post-heating. DISCUSSION AND CONCLUSION: It was concluded that heat-induced testicular damage causes both loss and relocation of SP22 on the sperm membrane. Future studies are warranted to determine the diagnostic value of these findings.

Characterisation of the testicular transcriptome in stallions with age-related testicular degeneration.

BACKGROUND: Age-related testicular degeneration can be defined as the progressive deterioration of the testis that typically occurs in middle-aged or older males and that leads to diminished testicular function and subfertility. In the equine breeding industry, genetically valuable males maintain their value as breeding animals well into old age. Because testicular degeneration is common in middle-aged and older stallions, the disease often has a significant negative impact on a stallion's breeding career and leads to economic losses in the horse breeding industry. OBJECTIVE: Because testicular degeneration is a tissue autologous disease in the horse, the objective of this study was to use whole-transcriptome sequencing to compare the testicular transcriptomes of normal, fertile stallions to those of stallions affected by age-related testicular degeneration in order to better understand the pathophysiology of the disease. STUDY DESIGN: Cross sectional. METHODS: Testicular tissue samples from clinical castrations or euthanasia were collected from normal healthy (n = 3) or older subfertile (n = 4) stallions. Samples were processed and sequenced on an Illumina HiSeq™ 2000 Sequencing System. Bioinformatic analysis of the data was performed in R/RStudio, and the transcriptomes were compared between the two groups. Genes were considered to be differentially expressed between healthy and diseased tissue if they demonstrated at least a ±1.5× fold change difference and had a false discovery rate-adjusted P value <0.05. Gene ontology analysis was performed using Ingenuity® IPA. RESULTS: Analyses of differential expression of individual genes, as well as computer-based gene ontology analysis, identified upregulation of cytokine-mediated inflammatory pathways in testes from stallions affected with testicular degeneration. This upregulation of inflammation was associated with upregulation of cell survival pathways, inhibition of apoptotic pathways and increases in collagen formation. MAIN LIMITATIONS: There are unavoidable confounding factors (e.g. differences in breed, management, environment, age) that could create non disease-related genetic variation between our normal and affected samples. In addition, there are practical limitations to applying computer-based gene ontology analysis to equine samples. Gene ontology software relies on published information (mostly non-equine), and some biological processes (e.g. apoptosis and inflammation) are more commonly studied than others and so are over-represented in the literature and therefore more likely to be identified by computer algorithms. Caution should be taken when interpreting the data, as alterations in gene expression can be the cause of disease processes or can be the result of disease processes. CONCLUSIONS: These results suggest that chronic, low-grade inflammation may be involved in the pathophysiology of age-related testicular degeneration in stallions.

Objective Characterization of Activity, Sleep, and Circadian Rhythm Patterns Using a Wrist-Worn Actigraphy Sensor: Insights Into Posttraumatic Stress Disorder.

BACKGROUND: Wearables have been gaining increasing momentum and have enormous potential to provide insights into daily life behaviors and longitudinal health monitoring. However, to date, there is still a lack of principled algorithmic framework to facilitate the analysis of actigraphy and objectively characterize day-by-day data patterns, particularly in cohorts with sleep problems. OBJECTIVE: This study aimed to propose a principled algorithmic framework for the assessment of activity, sleep, and circadian rhythm patterns in people with posttraumatic stress disorder (PTSD), a mental disorder with long-lasting distressing symptoms such as intrusive memories, avoidance behaviors, and sleep disturbance. In clinical practice, these symptoms are typically assessed using retrospective self-reports that are prone to recall bias. The aim of this study was to develop objective measures from patients' everyday lives, which could potentially considerably enhance the understanding of symptoms, behaviors, and treatment effects. METHODS: Using a wrist-worn sensor, we recorded actigraphy, light, and temperature data over 7 consecutive days from three groups: 42 people diagnosed with PTSD, 43 traumatized controls, and 30 nontraumatized controls. The participants also completed a daily sleep diary over 7 days and the standardized Pittsburgh Sleep Quality Index questionnaire. We developed a novel approach to automatically determine sleep onset and offset, which can also capture awakenings that are crucial for assessing sleep quality. Moreover, we introduced a new intuitive methodology facilitating actigraphy exploration and characterize day-by-day data across 49 activity, sleep, and circadian rhythm patterns. RESULTS: We demonstrate that the new sleep detection algorithm closely matches the sleep onset and offset against the participants' sleep diaries consistently outperforming an existing open-access widely used approach. Participants with PTSD exhibited considerably more fragmented sleep patterns (as indicated by greater nocturnal activity, including awakenings) and greater intraday variability compared with traumatized and nontraumatized control groups, showing statistically significant (P<.05) and strong associations (|R|>0.3). CONCLUSIONS: This study lays the foundation for objective assessment of activity, sleep, and circadian rhythm patterns using passively collected data from a wrist-worn sensor, facilitating large community studies to monitor longitudinally healthy and pathological cohorts under free-living conditions. These findings may be useful in clinical PTSD assessment and could inform therapy and monitoring of treatment effects.

Night-time rumination in PTSD: development and validation of a brief measure.

Background: Pre-sleep cognitive activity and arousal have long been implicated in the maintenance of insomnia. However, despite high comorbidity between insomnia and posttraumatic stress disorder (PTSD), pre-sleep thoughts in PTSD and their associations with disturbed sleep, have not yet been investigated. Objective: This study presents the development and preliminary validation of a brief self-report measure of the content of trauma-related pre-sleep thoughts: the Trauma Thoughts before Sleep Inventory (TTSI). Methods: Participants (N = 285) were recruited online into five groups: three groups with clinical symptoms, 1) PTSD; 2) depression without PTSD; 3) insomnia without depression or PTSD; and two healthy control groups 4) nontrauma-exposed controls; 5) trauma-exposed controls. The questionnaire was administered at baseline, and for a subsample (n = 157) again one week later to assess test-retest reliability. At baseline, participants also completed questionnaires of sleep quality, PTSD and depression symptoms, and insomnia-related thoughts. Results: The TTSI had good reliability and validity; it discriminated participants with PTSD from those with depression and insomnia, those with depression from insomnia, and correlated with existing measures of pre-sleep thoughts, self-reported pre-sleep arousal and poor sleep. Conclusions: The results support the utility of the TTSI for measuring thoughts that keep people with PTSD awake, although replication in an independent clinical sample is required.

Antecedentes: La actividad cognitiva y la excitación previa al sueño han estado implicadas durante mucho tiempo en el mantenimiento del insomnio. Sin embargo, a pesar de la alta comorbilidad entre el insomnio y el trastorno de estrés postraumático (TEPT), los pensamientos previos al sueño en el trastorno de estrés postraumático y sus asociaciones con trastornos del sueño, aún no se han investigado.Objetivo: Este estudio presenta el desarrollo y la validación preliminar de una escala breve de autoreporte del contenido de los pensamientos previos al sueño relacionados con el trauma: El Inventario de Pensamientos Traumáticos Previos al Sueño (IPTPS).Métodos: Los participantes (N = 285) fueron reclutados en línea y se dividieron en cinco grupos: tres grupos con síntomas clínicos, 1) TEPT; 2) depresión sin TEPT; 3) insomnio sin depresión o TEPT; y dos grupos de control sanos 4) controles no expuestos a traumas; 5) Controles expuestos al trauma. El cuestionario se administró al inicio del estudio y para una submuestra (n = 157) nuevamente una semana después, para evaluar la confiabilidad test-retest. Al inicio, los participantes también completaron cuestionarios de calidad del sueño, trastorno de estrés postraumático y depresión, y pensamientos relacionados con el insomnio.Resultados: El IPTPS tuvo buena confiabilidad y validez; discriminó a los participantes con TEPT de aquellos con depresión e insomnio, aquellos con depresión de los con insomnio y se correlacionó con las medidas existentes de los pensamientos previos al sueño y auto-reporte de activación previa al sueño y mala calidad de sueño.Conclusiones: Los resultados apoyan la utilidad del IPTPS para medir pensamientos que mantienen despiertas a las personas con TEPT, aunque se requiere la replicación en una muestra clínica independiente.

Differential Effects of Poor Recall and Memory Disjointedness on Trauma Symptoms.

Clinical theories of posttraumatic stress disorder (PTSD) suggest that trauma memories are disorganized. In the present study, we examined how trauma-film exposure affects two aspects of memory disorganization, poor memory recall and memory disjointedness, and their relationship to PTSD-like symptoms. In Session 1, 90 healthy participants were exposed to a trauma (n = 60) or a neutral film (n = 30). Cognitive processing styles, memory characteristics, and intrusive memories of the film were assessed. The trauma-film group reported greater memory disjointedness of the worst moments of the film but better memory recall of the film than the neutral-film group. In the trauma-film group, cognitive processing and memory disjointedness were related to intrusive memories and PTSD-like symptoms in the week after film exposure. Memory disjointedness but not poor memory recall mediated the relationship between cognitive processing and intrusions. The findings suggest that different aspects of memory disorganization need to be distinguished to explain PTSD symptoms.

Appraisal Biases About Strangers in Posttraumatic Stress Disorder.

Cognitive theories of posttraumatic stress disorder (PTSD) suggest that negative appraisals play an important role in the development and maintenance of PTSD. It has not yet been examined experimentally how people with PTSD appraise strangers. Twenty-two trauma survivors with PTSD and 26 non-traumatised controls completed a person impression updating task. There was no group difference in initial kindness ratings of people shown in the photos. The PTSD group changed their ratings more when receiving negative compared to positive information about the person, relative to controls. When incongruent information was subsequently presented, the PTSD group tended to be more influenced by negative information than positive information, relative to congruent information, than controls. The findings suggest selective attention to negative information about others in PTSD.

Effects of psychotherapies for posttraumatic stress disorder on sleep disturbances: Results from a randomized clinical trial.

The effectiveness and mechanisms of psychotherapies for posttraumatic stress disorder (PTSD) in treating sleep problems is of interest. This study compared the effects of a trauma-focused and a non-trauma-focused psychotherapy on sleep, to investigate whether 1) sleep improves with psychotherapy for PTSD; 2) the degree of sleep improvement depends on whether the intervention is trauma or nontrauma-focused; 3) the memory-updating procedure in cognitive therapy for PTSD (CT-PTSD) is associated with sleep improvements; 4) initial sleep duration affects PTSD treatment outcome; and 5) which symptom changes are associated with sleep duration improvements. Self-reported sleep was assessed during a randomized controlled trial (Ehlers et al., 2014) comparing CT-PTSD (delivered weekly or intensively over 7-days) with emotion-focused supportive therapy, and a waitlist. Sleep duration was reported daily in sleep diaries during intensive CT-PTSD. CT-PTSD led to greater increases in sleep duration (55.2 min) and reductions in insomnia symptoms and nightmares than supportive therapy and the waitlist. In intensive CT-PTSD, sleep duration improved within 7 days, and sleep diaries indicated a 40-min sleep duration increase after updating trauma memories. Initial sleep duration was not related to CT-PTSD treatment outcome when initial PTSD symptom severity was controlled. The results suggest that trauma-focused psychotherapy for PTSD is more effective than nontrauma-focused therapy in improving self-reported sleep, and that CT-PTSD can still be effective in the presence of reduced sleep duration.

The influence of age, antral follicle count and diestrous ovulations on estrous cycle characteristics of mares.

Reproductive aging must be well understood to optimize the reproductive management of older mares and to predict their reproductive life-span. The objectives of this study were to: 1) examine age-related differences in follicular dynamics, endocrine profiles, and primordial follicle counts, 2) evaluate the influence of antral follicle count (AFC) on age-related changes in follicular parameters, and 3) determine the influence of diestrous ovulations on the estrous cycle. Young (3-8yr; n = 10), middle-aged (9-18 yr; n = 16), and old (>18 yr; n = 19) light horse mares were examined with transrectal ultrasonography to monitor follicular growth over two consecutive estrous cycles. Jugular blood samples were taken and plasma progesterone and FSH concentrations were determined by an enzyme immunoassay and radioimmunoassay, respectively. Both interovulatory intervals and follicular phases were longer and the day of follicle deviation occurred later in aged mares. Furthermore, older mares had a tendency to ovulate smaller follicles. Neither follicular growth rate, the number of ovulations nor the length of luteal phase was influenced by mare age. Interestingly, as mare age increased, mares with low AFC had longer interovulatory intervals and follicular phases than mares with medium or high AFC. In addition, the number of primordial follicles declined with an increase in mare age but varied considerably between mares of the same age. Progesterone concentrations were positively influenced by age, whereas FSH concentrations were not, despite that FSH concentrations appeared higher in aged mares during the follicular phase. Estrous cycles with a diestrous ovulation had a longer interovulatory interval as well as a longer follicular and luteal phase while day of deviation occurred later. Progesterone concentrations were significantly higher on day 14 and 16 in estrous cycles with a diestrous ovulation than without a diestrous ovulation. In conclusion, aging in mares is associated with changes in follicular parameters which in turn are closely linked to differences in antral follicle count suggesting a relationship with ovarian reserve. Therefore, determination of antral follicle counts in aged mares can provide valuable information about the reproductive aging process. Finally, diestrous ovulations have a significant influence on different estrous cycle parameters.

Our current understanding of the pathophysiology of equine endometritis with an emphasis on breeding-induced endometritis.

Equine endometritis is characterized by the expression of pro- and anti- inflammatory cytokines and an influx of polymorphonuclear neutrophils (PMNs) into the uterus. Mares resistant to persistent endometritis clear the inflammation within 24-36 h after exposure to microorganisms or semen. These mares have a rapid increase of pro-inflammatory cytokines and an upregulation of inflammatory modulating cytokines within 6h after exposure to inflammatory challenge. In conjunction with effective uterine contractions, these events are believed to be responsible for the transient nature of the inflammation. In contrast, mares that are susceptible to persistent endometritis fail to clear the inflammation in a timely fashion. They have an imbalanced endometrial mRNA expression of pro- and anti-inflammatory cytokines, and have also been shown to suffer from an accumulation of intraluminal nitric oxide, which may be related to impaired myoelectrical activity and delayed uterine clearance. As a consequence, these mares establish a chronic inflammation, which interferes with the establishment of pregnancy. Recent studies on endometrial cytokine expression in resistant and susceptible mares have revealed information that suggest an underlying immunologic basis for susceptibility to persistent endometritis. However, the inflammatory pathways have yet not been fully studied, and a relationship between cytokine expression, nitric oxide, and myometrial contractions has not been established. In addition, seminal plasma has been shown to modulate breeding induced inflammation, but the molecular basis of the modulation is not understood. A holistic approach appears to be needed to better understand the characteristics of inflammatory pathways and ultimately the pathophysiology of the disease.

Imagery Rescripting for Obsessive Compulsive Disorder: A single case experimental design in 12 cases.

BACKGROUND AND OBJECTIVES: Some individuals with Obsessive Compulsive Disorder (OCD) may experience recurrent intrusive distressing images, which may be emotionally linked to past aversive memories. Our aim was to investigate whether Imagery Rescripting was an effective intervention for such individuals with OCD. METHOD: Twelve cases who experienced intrusive distressing images are presented in a A1BA2CA3 single case experimental design. After a baseline of symptom monitoring (A1), participants had a control intervention of talking about the memory related image (B), followed by symptom monitoring (A2), a single session of Imagery Rescripting (C) and further monitoring for up to 3 months (A3). RESULTS: Minimal change was seen following the control intervention. However, at 3 months following ImRs, there was a drop in the Yale-Brown Obsessive Compulsive Scale, with a decrease from a mean of 24.1 to 10.7. Reliable improvement was achieved in 9 out of the 12 cases and clinically significant change in 7 out of 12 at 3-month follow up. The limitations are that all cases were selected on the basis that they had an aversive memory linked to their imagery. CONCLUSIONS: Imagery Rescripting is a promising therapeutic technique for OCD as an adjunct to CBT where intrusive images are linked to aversive memories.

Internet-delivered cognitive therapy for social anxiety disorder: a development pilot series.

BACKGROUND: Randomized controlled trials have established that individual cognitive therapy based on the Clark and Wells (1995) model is an effective treatment for social anxiety disorder that is superior to a range of alternative psychological and pharmacological interventions. Normally the treatment involves up to 14 weekly face-to-face therapy sessions. AIM: To develop an internet based version of the treatment that requires less therapist time. METHOD: An internet-delivered version of cognitive therapy (iCT) for social anxiety disorder is described. The internet-version implements all key features of the face-to-face treatment; including video feedback, attention training, behavioural experiments, and memory focused techniques. Therapist support is via a built-in secure messaging system and by brief telephone calls. A cohort of 11 patients meeting DSM-IV criteria for social anxiety disorder worked through the programme and were assessed at pretreatment and posttreatment. RESULTS: No patients dropped out. Improvements in social anxiety and related process variables were within the range of those observed in randomized controlled trials of face-to-face CT. Nine patients (82%) were classified as treatment responders and seven (64%) achieved remission status. Therapist time per patient was only 20% of that in face-to-face CT. CONCLUSIONS: iCT shows promise as a way of reducing therapist time without compromising efficacy. Further evaluation of iCT is ongoing.

Inflammatory responses to induced infectious endometritis in mares resistant or susceptible to persistent endometritis.

BACKGROUND: The objective of the study was to evaluate the gene expression of inflammatory cytokines (interleukin [IL]-1ß, IL-6, IL-8, IL-10, tumor necrosis factor [TNF]-α, IL-1 receptor antagonist [ra] and serum amyloid A (SAA) in endometrial tissue and circulating leukocytes in response to uterine inoculation of 105 colony forming units (CFU) Escherichia coli in mares. Before inoculation, mares were classified as resistant or susceptible to persistent endometritis based on their uterine inflammatory response to infusion of 109 killed spermatozoa and histological assessment of the endometrial quality. Endometrial biopsies were obtained 3, 12, 24 and 72 hours (h) after bacterial inoculation and blood samples were obtained during the 7 day period post bacterial inoculation. Expression levels of cytokines and SAA were determined by quantitative real-time reverse transcriptase PCR (qRT-PCR). RESULTS: Compared to levels in a control biopsy (obtained in the subsequent estrous), resistant mares showed an up-regulation of IL-1ß, IL-6, IL-8 and TNF-α at 3 h after E. coli inoculation, while susceptible mares showed increased gene expression of IL-6 and IL-1ra. Susceptible mares had a significant lower gene expression of TNF-α,IL-6 and increased expression of IL-1ra 3 h after E. coli inoculation compared to resistant mares. Susceptible mares showed a sustained and prolonged inflammatory response with increased gene expression levels of IL-1ß, IL-8, IL-1ra and IL-1ß:IL-1ra ratio throughout the entire study period (72 h), whereas levels in resistant mares returned to estrous control levels by 12 hours. Endometrial mRNA transcripts of IL-1ß and IL-1ra were significantly higher in mares with heavy uterine bacterial growth compared to mares with no/mild growth.All blood parameters were unaffected by intrauterine E. coli infusion, except for a lower gene expression of IL-10 at 168 h and an increased expression of IL-1ra at 48 h observed in susceptible mares compared to resistant mares. CONCLUSIONS: The current investigation suggests that endometrial mRNA transcripts of pro-inflammatory cytokines in response to endometritis are finely regulated in resistant mares, with initial high expression levels followed by normalization within a short period of time. Susceptible mares had a prolonged expression of pro-inflammatory cytokines, supporting the hypothesis that an unbalanced endometrial gene expression of inflammatory cytokines might play an important role in the pathogenesis of persistent endometritis.