A novel murine model mimicking male genital Neisseria species infection using Neisseria musculi†.

With ~78 million cases yearly, the sexually transmitted bacterium Neisseria gonorrhoeae is an urgent threat to global public health due to continued emergence of antimicrobial resistance. In the male reproductive tract, untreated infections may cause permanent damage, poor sperm quality, and subsequently subfertility. Currently, few animal models exist for N. gonorrhoeae infection, which has strict human tropism, and available models have limited translatability to human disease. The absence of appropriate models inhibits the development of vital new diagnostics and treatments. However, the discovery of Neisseria musculi, a mouse oral cavity bacterium, offers much promise. This bacterium has already been used to develop an oral Neisseria infection model, but the feasibility of establishing urogenital gonococcal models is unexplored. We inoculated mice via the intrapenile route with N. musculi. We assessed bacterial burden throughout the male reproductive tract, the systemic and tissue-specific immune response 2-weeks postinfection, and the effect of infection on sperm health. Neisseria musculi was found in penis (2/5) and vas deferens (3/5) tissues. Infection altered immune cell counts: CD19+ (spleen, lymph node, penis), F4/80+ (spleen, lymph node, epididymus), and Gr1+ (penis) compared with noninfected mice. This culminated in sperm from infected mice having poor viability, motility, and morphology. We hypothesize that in the absence of testis infection, infection and inflammation in other reproductive is sufficient to damage sperm quality. Many results herein are consistent with outcomes of gonorrhoea infection, indicating the potential of this model as a tool for enhancing the understanding of Neisseria infections of the human male reproductive tract.

Haematopoietic innate interleukin 17A production drives immunopathology in female mouse genital Chlamydia muridarum infection.

Chlamydia trachomatis infection is the leading cause of bacterial urogenital infection and has been demonstrated to drive inflammation and scarring of the reproductive tract. Recent studies have identified key triggers of proinflammatory adaptive immune responses driven by innate leukocytes and epithelia driving immunopathology. Utilizing chimeric mouse models, we investigated the definitive source and role of IL17 and IL17 signalling receptors during early Chlamydia muridarum infection of the female urogenital tract. Bone marrow transplants from wild-type (WT) and IL17A-/- mice to recipients demonstrated equivocal infection kinetics in the reproductive tract, but interestingly, adoptive transfer of IL17A-/- immune cells to WT recipients resulted in no infertility, suggesting a haematopoietic (as opposed to tissue) source of IL17 driving immunopathology. To further delineate the role of IL17 in immunopathology, we infected WT and IL17 receptor A (IL17RA)-/- female mice and observed a significant reduction in immunopathology in IL17RA-/- mice. WT bone marrow transplants to IL17RA-/- recipient mice prevented hydrosalpinx, suggesting signalling through IL17RA drives immunopathology. Furthermore, early chemical inhibition of IL17 signalling significantly reduced hydrosalpinx, suggesting IL17 acts as an innate driver of disease. Early during the infection, IL17 was produced by γδ T cells in the cervico-vagina, but more importantly, by neutrophils at the site of infertility in the oviducts. Taken together, these data suggest innate production of IL17 by haematopoietic leukocytes drives immunopathology in the epithelia during early C. muridarum infection of the female reproductive tract.

IgG exacerbates genital chlamydial pathology in females by enhancing pathogenic CD8+ T cell responses.

Chlamydia trachomatis infections are an important sexually transmitted infection that can lead to inflammation, scarring and hydrosalpinx/infertility. However, infections are commonly clinically asymptomatic and do not receive treatment. The underlying cause of asymptomatic immunopathology remains unknown. Here, we demonstrate that IgG produced during male infection enhanced the incidence of immunopathology and infertility in females. Human endocervical cells expressing the neonatal Fc Receptor (FcRn) increased translocation of human IgG-opsonized C. trachomatis. Using total IgG purified from infected male mice, we opsonized C. muridarum and then infected female mice, mimicking sexual transmission. Following infection, IgG-opsonized Chlamydia was found to transcytose the epithelial barrier in the uterus, where it was phagocytosed by antigen-presenting cells (APCs) and trafficked to the draining lymph nodes. APCs then expanded both CD4+ and CD8+ T cell populations and caused significantly more infertility in female mice infected with non-opsonized Chlamydia. Enhanced phagocytosis of IgG-opsonized Chlamydia significantly increased pro-inflammatory signalling and T cell proliferation. As IgG is transcytosed by FcRn, we utilized FcRn-/- mice and observed that shedding kinetics of Chlamydia were only affected in FcRn-/- mice infected with IgG-opsonized Chlamydia. Depletion of CD8+ T cells in FcRn-/- mice lead to a significant reduction in the incidence of infertility. Taken together, these data demonstrate that IgG seroconversion during male infection can amplify female immunopathology, dependent on FcRn transcytosis, APC differentiation and enhanced CD8 T cell responses.

Lactobacillus rhamnosus dampens cytokine and chemokine secretion from primary human nasal epithelial cells infected with rhinovirus.

AIMS: To investigate the effect of Lactobacillus rhamnosus on viral replication and cellular response to human rhinovirus (HRV) infection, including the secretion of antiviral and inflammatory mediators from well-differentiated nasal epithelial cells (WD-NECs). METHODS AND RESULTS: The WD-NECs from healthy adult donors (N = 6) were cultured in vitro, exposed to different strains of L. rhamnosus (D3189, D3160, or LB21), and infected with HRV (RV-A16) after 24 h. Survival and adherence capacity of L. rhamnosus in a NEC environment were confirmed using CFSE-labelled isolates, immunofluorescent staining, and confocal microscopy. Shed virus and viral replication were quantified using TCID50 assays and RT-qPCR, respectively. Cytotoxicity was measured by lactate dehydrogenase (LDH) activity. Pro-inflammatory mediators were measured by multiplex immunoassay, and interferon (IFN)-λ1/3 was measured using a standard ELISA kit. Lactobacillus rhamnosus was able to adhere to and colonize WD-NECs prior to the RV-A16 infection. Lactobacillus rhamnosus did not affect shed RV-A16, viral replication, RV-A16-induced IFN-λ1/3 production, or LDH release. Pre-exposure to L. rhamnosus, particularly D3189, reduced the secretion of RV-A16-induced pro-inflammatory mediators by WD-NECs. CONCLUSIONS: These findings demonstrate that L. rhamnosus differentially modulates RV-A16-induced innate inflammatory immune responses in primary NECs from healthy adults.

Prophylactic and therapeutic vaccination protects sperm health from Chlamydia muridarum-induced abnormalities.

Chlamydia is the most common bacterial sexually transmitted infection worldwide and it is widely acknowledged that controlling the rampant community transmission of this infection requires vaccine development. In this study, for the first time, we elucidate the long-term response to male mouse chlamydial vaccination with chlamydial major outer membrane protein (MOMP) and ISCOMATRIX (IMX) both prophylactically and in a novel therapeutic setting. Vaccination significantly reduced and, in some cases, cleared chlamydial burden from the prostates, epididymides, and testes, which correlates with high IgG and IgA tires in tissues and serum. Important markers of sperm health and fertility were protected including sperm motility and proteins associated with fertility in men. Within splenocytes, expression of IFNγ, TNFα, IL17, IL13, IL10, and TGFß were changed by both infection and vaccination within CD4 and CD8 T cells and regulatory T cells. Within the testicular tissue, phenotypic and concentration changes were observed in macrophages and T cells (resident and transitory). This revealed some pathogenic phenotypes associated with infection and critically that vaccination allows maintenance of testicular homeostasis, likely by preventing significant influx of CD4 T cells and promoting IL10 production. Finally, we demonstrated the testes contained immature (B220+) B cells and mature (CD138+) Chlamydia-specific plasma cells. Thus, through vaccination, we can maintain the healthy function of the testes, which is vital to protection of male fertility.

Pathogenic NKT cells attenuate urogenital chlamydial clearance and enhance infertility.

Urogenital chlamydial infections continue to increase with over 127 million people affected annually, causing significant economic and public health pressures. While the role of traditional MHCI and II peptide presentation is well defined in chlamydial infections, the role of lipid antigens in immunity remains unclear. Natural killer (NK) T cells are important effector cells that recognize and respond to lipid antigens during infections. Chlamydial infection of antigen-presenting cells facilitates presentation of lipid on the MHCI-like protein, CD1d, which stimulates NKT cells to respond. During urogenital chlamydial infection, wild-type (WT) female mice had significantly greater chlamydial burden than CD1d-/- (NKT-deficient) mice, and had significantly greater incidence and severity of immunopathology in both primary and secondary infections. WT mice had similar vaginal lymphocytic infiltrate, but 59% more oviduct occlusion compared to CD1d-/- mice. Transcriptional array analysis of oviducts day 6 post-infection revealed WT mice had elevated levels of Ifnγ (6-fold), Tnfα (38-fold), Il6 (2.5-fold), Il1ß (3-fold) and Il17a (6-fold) mRNA compared to CD1d-/- mice. In infected females, oviduct tissues had an elevated infiltration of CD4+ -invariant NKT (iNKT) cells, however, iNKT-deficient Jα18-/- mice had no significant differences in hydrosalpinx severity or incidence compared to WT controls. Lipid mass spectrometry of surface-cleaved CD1d in infected macrophages revealed an enhancement of presented lipids and cellular sequestration of sphingomyelin. Taken together, these data suggest an immunopathogenic role for non-invariant NKT cells in urogenital chlamydial infections, facilitated by lipid presentation via CD1d via infected antigen-presenting cells.

Intolerance of Uncertainty, Negative Reinforcement Alcohol Use Motives, and Hazardous Drinking in College Students with Clinically Elevated Worry.

Background: Hazardous drinking has remained high for college students in recent years, and individuals who endorse drinking to cope with emotional distress or to conform socially report higher levels of alcohol use. Intolerance of uncertainty, a core process underlying generalized anxiety disorder, has been linked to negative reinforcement drinking motives; however, no research, to date, has examined the role of intolerance of uncertainty in terms of alcohol use motives and hazardous drinking among individuals with generalized anxiety disorder. Objective: The current study was designed to examine the relationships between intolerance of uncertainty, coping and conformity alcohol use motives, and hazardous drinking in an analogue generalized anxiety disorder sample. Methods: Participants were 323 college students (Mage = 19.25, SD = 2.23, Range = 18-40) who endorsed past-year alcohol use and clinically elevated levels of worry. Self-report measures were completed online for course credit. Results: Partially consistent with our hypotheses, uncertainty paralysis predicted greater levels of coping motives, but not conformity motives. Desire for predictability did not predict either drinking motive. Mediation analyses revealed that there was a significant indirect effect of uncertainty paralysis on more hazardous drinking through greater coping motives. Conclusion: Overall, these findings highlight the potential utility of targeting behavioral inhibition due to uncertainty to reduce unhealthy coping via alcohol use and subsequent hazardous alcohol use.

Implementation of a structured practical activity to analyse student healthcare worker perceptions and compliance with prescribed infection control procedures.

BACKGROUND: Non-compliance with infection control guidelines has been reported within healthcare settings. Infection control education in undergraduate healthcare education programs forms a critical component in preparing student healthcare workers for vocational roles. METHODS: Clinical sciences students (nutrition science, paramedicine, pharmacy, podiatry, optometry studying for qualifications recognised by the Australian Health Practitioner Regulation Agency) self-reported hygiene perceptions and practices and collected microbiological swabs from personal or medical equipment items before and after recommended disinfection procedures. RESULTS: Cultivable microorganisms were isolated from 95% of student medical equipment items. Disinfection significantly reduced microbial growth on student medical equipment items (P < 0.05). CONCLUSIONS: Student perceptions of infection control procedures do not always correlate with infection control practice. Infection control education of undergraduate healthcare students requires ongoing assessment to ensure successful translation into clinical practice.

Chronic testicular Chlamydia muridarum infection impairs mouse fertility and offspring development†.

With approximately 131 million new genital tract infections occurring each year, Chlamydia is the most common sexually transmitted bacterial pathogen worldwide. Male and female infections occur at similar rates and both cause serious pathological sequelae. Despite this, the impact of chlamydial infection on male fertility has long been debated, and the effects of paternal chlamydial infection on offspring development are unknown. Using a male mouse chronic infection model, we show that chlamydial infection persists in the testes, adversely affecting the testicular environment. Infection increased leukocyte infiltration, disrupted the blood:testis barrier and reduced spermiogenic cell numbers and seminiferous tubule volume. Sperm from infected mice had decreased motility, increased abnormal morphology, decreased zona-binding capacity, and increased DNA damage. Serum anti-sperm antibodies were also increased. When both acutely and chronically infected male mice were bred with healthy female mice, 16.7% of pups displayed developmental abnormalities. Female offspring of chronically infected sires had smaller reproductive tracts than offspring of noninfected sires. The male pups of infected sires displayed delayed testicular development, with abnormalities in sperm vitality, motility, and sperm-oocyte binding evident at sexual maturity. These data suggest that chronic testicular Chlamydia infection can contribute to male infertility, which may have an intergenerational impact on sperm quality.

Hematogenous dissemination of Chlamydia muridarum from the urethra in macrophages causes testicular infection and sperm DNA damage†.

The incidence of Chlamydia infection, in both females and males, is increasing worldwide. Male infections have been associated clinically with urethritis, epididymitis, and orchitis, believed to be caused by ascending infection, although the impact of infection on male fertility remains controversial. Using a mouse model of male chlamydial infection, we show that all the major testicular cell populations, germ cells, Sertoli cells, Leydig cells, and testicular macrophages can be productively infected. Furthermore, sperm isolated from vas deferens of infected mice also had increased levels of DNA damage as early as 4 weeks post-infection. Bilateral vasectomy, prior to infection, did not affect the chlamydial load recovered from testes at 2, 4, and 8 weeks post-infection, and Chlamydia-infected macrophages were detectable in blood and the testes as soon as 3 days post-infection. Partial depletion of macrophages with clodronate liposomes significantly reduced the testicular chlamydial burden, consistent with a hematogenous route of infection, with Chlamydia transported to the testes in infected macrophages. These data suggest that macrophages serve as Trojan horses, transporting Chlamydia from the penile urethra to the testes within 3 days of infection, bypassing the entire male reproductive tract. In the testes, infected macrophages likely transfer infection to Leydig, Sertoli, and germ cells, causing sperm DNA damage and impaired spermatogenesis.

Detection of chlamydia infection within human testicular biopsies.

STUDY QUESTION: Can Chlamydia be found in the testes of infertile men? SUMMARY ANSWER: Chlamydia can be found in 16.7% of fresh testicular biopsies and 45.3% of fixed testicular biopsies taken from a selection of infertile men. WHAT IS KNOWN ALREADY: Male chlamydial infection has been understudied despite male and female infections occurring at similar rates. This is particularly true of asymptomatic infections, which occur in 50% of cases. Chlamydial infection has also been associated with increased sperm DNA damage and reduced male fertility. STUDY DESIGN, SIZE, DURATION: We collected diagnostic (fixed, n = 100) and therapeutic (fresh, n = 18) human testicular biopsies during sperm recovery procedures from moderately to severely infertile men in a cross-sectional approach to sampling. PARTICIPANTS/MATERIALS, SETTING, METHODS: The diagnostic and therapeutic biopsies were tested for Chlamydia-specific DNA and protein, using real-time PCR and immunohistochemical approaches, respectively. Serum samples matched to the fresh biopsies were also assayed for the presence of Chlamydia-specific antibodies using immunoblotting techniques. MAIN RESULTS AND THE ROLE OF CHANCE: Chlamydial major outer membrane protein was detected in fixed biopsies at a rate of 45.3%. This was confirmed by detection of chlamydial DNA and TC0500 protein (replication marker). C. trachomatis DNA was detected in fresh biopsies at a rate of 16.7%, and the sera from each of these three positive patients contained C. trachomatis-specific antibodies. Overall, C. trachomatis-specific antibodies were detected in 72.2% of the serum samples from the patients providing fresh biopsies, although none of the patients were symptomatic nor had they reported a previous sexually transmitted infection diagnosis including Chlamydia. LIMITATIONS, REASONS FOR CAUTION: No reproductively healthy male testicular biopsies were tested for the presence of Chlamydia DNA or proteins or Chlamydia-specific antibodies due to the unavailability of these samples. WIDER IMPLICATIONS FOR THE FINDINGS: Application of Chlamydia-specific PCR and immunohistochemistry in this human male infertility context of testicular biopsies reveals evidence of a high prevalence of previously unrecognised infection, which may potentially have a pathogenic role in spermatogenic failure. STUDY FUNDING/COMPETING INTEREST(S): Funding for this project was provided by the Australian NHMRC under project grant number APP1062198. We also acknowledge assistance from the Monash IVF Group and Queensland Fertility Group in the collection of fresh biopsies, and the Monash Health and co-author McLachlan (declared equity interest) in retrieval and sectioning of fixed biopsies. E.M. declares an equity interest in the study due to financing of fixed biopsy sectioning. All other authors declare no conflicts of interest. TRIAL REGISTRATION NUMBER: N/A.

The role of anxiety sensitivity-physical concerns in terms of quit day withdrawal symptoms and cravings: A pilot test among smokers with asthma.

OBJECTIVE: The aim of the current study was to conduct secondary analyses of data collected from a study examining a self-guided quit attempt of smokers with and without asthma in order to examine the unique predictive ability of precessation (i.e., baseline) anxiety sensitivity physical concerns in terms of quit day withdrawal symptoms and cravings among abstinent smokers with asthma. METHODS: Participants were 24 regular cigarette smokers with asthma (50% female, Mage = 37.63 years, SD = 12.20) who participated in a self-guided-quit attempt and were able to maintain abstinence on their quit day. RESULTS: After controlling for the effects of the cognitive and social concerns domains of anxiety sensitivity, anxiety sensitivity-physical concerns significantly predicted greater quit day withdrawal symptoms (20.8% unique variance) and urges to smoke (38.0% unique variance). CONCLUSIONS: These findings suggest that smokers with asthma who fear anxiety-related sensations due to their feared physical consequences are more likely to experience intense withdrawal symptoms and desire to smoke at the beginning of a quit attempt. Clinically, smokers with higher levels of anxiety sensitivity physical concerns may benefit from smoking cessation interventions that specifically target anxiety sensitivity as well as prolonged use of nicotine replacement therapies to target withdrawal symptoms and cravings.

An Examination of the Association Between Emotion Reactivity and Distress Tolerance Among College Students.

The goal of the present investigation was to examine associations between emotion reactivity in global and specific distress tolerance domains. Participants were 113 undergraduates (65.5% female, 82.3% Caucasian; Mage = 20.0, SD = 4.02, range = 18-45 years) who completed a battery of self-report measures. As hypothesized, the results indicate that, after controlling for sex and negative affectivity, greater levels of emotion reactivity were significantly associated with lower levels of global distress tolerance (12.5% unique variance) and the four specific distress tolerance domains: tolerance (8.6% unique variance), absorption (11.4% unique variance), appraisal (8% unique variance), and regulation (8.1% unique variance). These findings suggest that individuals who tend to react more strongly to their emotions have greater difficulty tolerating emotional distress, and interventions that target emotion reactivity may be an effective way to improve distress tolerance and reduce the risk of developing psychopathology among undergraduates.

The role of pain catastrophizing in cyberchondria among emerging adults.

Chronic pain is a prevalent and debilitating health problem that often persists in the absence of an identifiable biomedical cause. Uncertainty regarding the origins of a pain problem may lead to pain catastrophizing and unfruitful healthcare seeking behaviors. Individuals with chronic pain often turn to the internet to identify possible causes or sources of their pain symptoms, thus affirming their pain experience. Despite evidence that pain catastrophizing amplifies distress about pain complaints, no studies have investigated whether it is associated with escalations in health anxiety that result from searching for online health information (i.e., cyberchondria). The aim of the present study was to investigate whether pain catastrophizing predicted variance in cyberchondria above and beyond health anxiety. Undergraduate students (N = 221, 70.6% female) completed questionnaires assessing pain catastrophizing, health anxiety, and cyberchondria. Results from hierarchical regression models indicated that pain catastrophizing predicted unique variance in four dimensions of cyberchondria: compulsion, distress, excessiveness, and reassurance. Findings are consistent with research highlighting the consequences of pain catastrophizing on myriad psychological outcomes. Individuals who catastrophize about pain experiences may be vulnerable to developing severe health anxiety from searching the internet for health information. Implications for research among individuals with chronic pain are discussed.

The indirect effect of positive affect in the relationship between trait mindfulness and emotion dysregulation.

Mindfulness is correlated with better emotion regulation and mindfulness training improves emotion regulation; however, mechanisms of these effects have not been widely studied. Therefore, the aim of the current study was to examine the indirect effect of greater mindfulness on fewer emotion regulation difficulties through positive affect. Participants were 219 undergraduate students (M age =19.20; 70.5% female; 91.5% Caucasian) who completed self-report measures for course credit. Results indicated a significant direct effect of mindfulness on emotion dysregulation (b = -1.71, t = -6.94, p < .001) and significant indirect effect of greater mindfulness on fewer emotion regulation problems through higher levels of positive affect (indirect effect =-0.50; SE = 0.13; 95% CI = -0.80 to -0.28). These results help to generate testable hypotheses for future mechanistic research in this area by suggesting that one way trait mindfulness may protect against emotion dysregulation could be by cultivating positive affect. Future research using longitudinal designs is needed to explore positive affect as a causal mechanism.

The role of mindfulness skills in terms of anxiety-related cognitive risk factors among college students with problematic alcohol use.

BACKGROUND: The aim of the current study was to examine the associations between the specific mindfulness skills of observing, describing, awareness, nonjudgment, and nonreactivity in terms of anxiety sensitivity (AS), distress tolerance (DT), and intolerance of uncertainty (IU) among college students with problematic alcohol use. METHODS: Participants were 202 (69.3% male; Mage = 18.96, SD = 2.24, range = 18-45 years) undergraduate college students with problematic alcohol use who completed self-report measures for course credit. RESULTS: Results indicated that after controlling for the effects of gender, smoking status, marijuana use status, and negative affectivity, greater use of the mindfulness skill of observing was associated with higher AS, greater describing was associated with lower AS and higher DT, greater nonjudgment was associated with lower AS and IU and higher DT, and greater nonreactivity was associated with increased DT. Awareness did not significantly predict any of the examined risk factors. CONCLUSIONS: These results suggest that specific mindfulness skills are associated with a greater tolerance of physiological, emotional, and uncertain states. An important next step will be to examine whether mindfulness skills are associated with decreased problematic alcohol use due to improvements in these anxiety-related risk factors.

Mind-Body Skills Training to Improve Distress Tolerance in Medical Students: A Pilot Study.

PROBLEM: Medical students face rigorous and stressful work environments, resulting in high rates of psychological distress. However, there has been a dearth of empirical work aimed at modifying risk factors for psychopathology among this at-risk group. Distress tolerance, defined as the ability to withstand emotional distress, is one factor that may be important in promoting psychological well-being in medical students. Thus, the aim of the current mixed-methods study was (a) to describe changes in facets of distress tolerance (i.e., emotional tolerance, absorption, appraisal, regulation) for medical students who completed a mind-body skills training group, and a no-intervention control group of students; (b) to examine the relationship between changes in psychological variables and changes in distress tolerance; and (c) to report students' perceptions of the mind-body group, with an emphasis on how the group may have affected personal and professional functioning due to improvements in distress tolerance. INTERVENTION: The mind-body program was an 11-week, 2-hour skills training group that focused on introducing, practicing, and processing mind-body skills such as biofeedback, guided imagery, relaxation, several forms of meditation (e.g., mindfulness), breathing exercises, and autogenic training. CONTEXT: Participants were 52 first- and second-year medical students (62.7% female, Mage = 23.45, SD = 1.51) who participated in a mind-body group or a no-intervention control group and completed self-report measures before and after the 11-week period. OUTCOME: Students in the mind-body group showed a modest improvement in all distress tolerance subscales over time (ΔM = .42-.53, p = .01-.03, d = .44-.53), whereas the control group showed less consistent changes across most subscales (ΔM = .11-.42, p = .10-.65, d = .01-.42). Students in the mind-body group qualitatively reported an improved ability to tolerate affective distress. Overall, improvements in psychological symptoms were associated with improvements in distress tolerance in the mind-body group but not in the control group. LESSONS LEARNED: These preliminary findings provide support for the notion that improving distress tolerance through mind-body skills training might serve to protect medical students from becoming functionally impaired by psychological distress. Thus, implementing mind-body skills training into medical school education may help to improve the psychological well-being of medical students. Future studies utilizing more methodologically rigorous designs are warranted.

Stability and antigenicity of Chlamydia muridarum major outer membrane protein antigen at body temperature.

Chlamydia is an obligate intracellular bacterial pathogen responsible for disease and infertility across multiple species. Currently vaccines are being studied to help reduce the prevalence of this disease. The main advantage of protein subunit vaccines is their high degree of safety although this is traded off with the requirement for multiple booster doses to achieve complete protection. Although in certain populations the booster dose can be difficult and costly to administer, development of delayed vaccine delivery techniques, such as a vaccine capsule, could be the solution to this problem. One of the main drawbacks in this technology is that the antigen must remain stable at body temperature (37 °C) until release is achieved. Here we elucidate the stability of a recombinant chlamydial major outer membrane protein (MOMP) antigen and assess its antigenic and immunogenic properties after subjecting the antigen to 37 °C for four to six weeks. Through in vitro and in vivo assessment we found that the aged chlamydial MOMP was able to produce equivalent humoral and cell-mediated immune responses when compared with the unaged vaccine. It was also found that vaccines formulated with the aged antigen conferred equivalent protection against a live infection challenge as the unaged antigen. Thus ageing chlamydial MOMP antigens at 37 °C for four to six weeks did not cause any significant structural or antigenic/immunogenic degradation and recombinant C. muridarum MOMP is suitable for use in a delayed vaccine delivery system.

Enhanced clearance of C. muridarum infection using azithromycin-loaded liposomes.

Chlamydia trachomatis is an intracellular bacterium which infects around 129 million people annually. Despite similar infection rates between sexes, most research investigating the effects of chlamydial infection on fertility has focused on females. There is now emerging evidence of a potential link between Chlamydia and impaired male fertility. The only treatments for chlamydial infection are antibiotics, with azithromycin (AZI) being one of the commonly used drugs. However, recent studies have suggested that optimizing the treatment regime is necessary, as higher concentrations of AZI may be required to effectively clear the infection in certain cell types, particularly testicular macrophages. To address this challenge, we have prepared liposomes consisting of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) loaded with AZI for clearing Chlamydia. These liposomes exhibited stability over time and were readily taken up by both macrophages and epithelial cells. Moreover, they demonstrated significant enhancement of chlamydial clearance in both cell types. In a mouse model, the drug-loaded liposomes cleared Chlamydia within the penile urethra more efficiently than the same dose of unencapsulated drug. Furthermore, the liposome-drug treatment showed significant protective effects on sperm motility and morphology, suggesting potential benefits in reducing sperm damage caused by the infection.

Brief adjunctive mindfulness-based cognitive therapy via Telehealth for anxiety during the COVID-19 pandemic.

BACKGROUND AND OBJECTIVES: While cognitive-behavioral therapy is a highly efficacious treatment for anxiety, additional research is needed to identify adjunctive interventions that may augment treatment outcome. DESIGN: In response to the COVID-19 pandemic, we conducted an open feasibility trial of brief (i.e., four 75- to 90-minute sessions) mindfulness-based cognitive therapy (MBCT) for anxiety via telehealth for patients (N = 23) receiving individual CBT at an outpatient specialty clinic. METHODS: Self-report measures of home practice compliance (weekly), intervention acceptability (post-intervention), mindfulness and self-compassion, anxiety and depressive symptoms, and transdiagnostic processes (pre- and post-intervention) were administered as part of routine clinical practice. RESULTS: Results indicated good retention and attendance rates, few technical difficulties, good home practice compliance, and high levels of perceived importance. Participants indicated that they would highly recommend the group to others and also recommended extending the group beyond four sessions. There were significant improvements in mindfulness and self-compassion and reductions in intolerance of uncertainty, anxiety sensitivity, distress intolerance, emotion dysregulation, and anxiety symptoms from pre- to post-intervention. CONCLUSIONS: Brief MBCT as an adjunctive treatment for anxiety via telehealth is feasible and acceptable, and shows promise in terms of engaging treatment targets and transdiagnostic processes and reducing anxiety symptoms.