Introducing carbon assimilation in yeasts using photosynthetic directed endosymbiosis.

Conversion of heterotrophic organisms into partially or completely autotrophic organisms is primarily accomplished by extensive metabolic engineering and laboratory evolution efforts that channel CO2 into central carbon metabolism. Here, we develop a directed endosymbiosis approach to introduce carbon assimilation in budding yeasts. Particularly, we engineer carbon assimilating and sugar-secreting photosynthetic cyanobacterial endosymbionts within the yeast cells, which results in the generation of yeast/cyanobacteria chimeras that propagate under photosynthetic conditions in the presence of CO2 and in the absence of feedstock carbon sources like glucose or glycerol. We demonstrate that the yeast/cyanobacteria chimera can be engineered to biosynthesize natural products under the photosynthetic conditions. Additionally, we expand our directed endosymbiosis approach to standard laboratory strains of yeasts, which transforms them into photosynthetic yeast/cyanobacteria chimeras. We anticipate that our studies will have significant implications for sustainable biotechnology, synthetic biology, and experimentally studying the evolutionary adaptation of an additional organelle in yeast.

Low-Frequency Raman Spectroscopy of Pure and Cocrystallized Mycophenolic Acid.

The aqueous solubility of solid-state pharmaceuticals can often be enhanced by cocrystallization with a coformer to create a binary cocrystal with preferred physical properties. Greater understanding of the internal and external forces that dictate molecular structure and intermolecular packing arrangements enables more efficient design of new cocrystals. Low-frequency (sub-200 cm-1) Raman spectroscopy experiments and solid-state density functional theory simulations have been utilized together to investigate the crystal lattice vibrations of mycophenolic acid, an immunosuppressive drug, in its pure form and as a cocrystal with 2,2'-dipyridylamine. The lattice vibrations primarily consist of large-amplitude translations and rotations of the crystal components, thereby providing insights into the critical intermolecular forces governing cohesion of the molecular solids. The simulations reveal that despite mycophenolic acid having a significantly unfavorable conformation in the cocrystal as compared to the pure solid, the cocrystal exhibits greater thermodynamic stability over a wide temperature range. The energetic penalty due to the conformational strain is more than compensated for by the strong intermolecular forces between the drug and 2,2'-dipyridylamine. Quantifying the balance of internal and external energy factors in cocrystal formation indicates a path forward in the development of future mycophenolic acid cocrystals.

Development of a brief support and education intervention for loved ones of veterans with posttraumatic stress disorder.

Clinical practice guidelines for managing posttraumatic stress disorder (PTSD) encourage incorporating loved ones into treatment, and supportive relationships can increase engagement in mental health care for veterans with PTSD. This study describes the iterative refinement, feasibility/acceptability testing, and national dissemination of a brief support and psychoeducation intervention for loved ones of veterans with PTSD. Loved ones (n = 181; range:1-11 per group) attended and qualitatively rated "PTSD 101 for Family and Friends: A Support and Education Workshop." Open-ended questions were used to gather data on suggestions for improvement and descriptions of helpful content, and the workshop was refined following participant and operational partner feedback using a quality improvement framework. Rating quantitative items on a 1-5 scale, participants found the overall quality (M = 4.76) and relevance (M = 4.82) to be excellent, noting they learned substantial new information (M = 4.45). Sense of support (M = 4.95), intentions to use the material (M = 4.87), PTSD self-efficacy (M = 4.41), and understanding of PTSD (M = 4.76) were rated favorably. Common themes among helpful elements were a sense of shared experience and optimism and increased knowledge of treatments/resources. Suggestions for improvement referenced logistics. Following continual stakeholder feedback and refinement, the workshop represents a novel method for providing loved ones with empirically supported psychoeducation, coping skills, and community. It is being disseminated by the Family Services Section of the Veterans Health Administration Office of Mental Health and Suicide Prevention.

Engineering artificial photosynthetic life-forms through endosymbiosis.

The evolutionary origin of the photosynthetic eukaryotes drastically altered the evolution of complex lifeforms and impacted global ecology. The endosymbiotic theory suggests that photosynthetic eukaryotes evolved due to endosymbiosis between non-photosynthetic eukaryotic host cells and photosynthetic cyanobacterial or algal endosymbionts. The photosynthetic endosymbionts, propagating within the cytoplasm of the host cells, evolved, and eventually transformed into chloroplasts. Despite the fundamental importance of this evolutionary event, we have minimal understanding of this remarkable evolutionary transformation. Here, we design and engineer artificial, genetically tractable, photosynthetic endosymbiosis between photosynthetic cyanobacteria and budding yeasts. We engineer various mutants of model photosynthetic cyanobacteria as endosymbionts within yeast cells where, the engineered cyanobacteria perform bioenergetic functions to support the growth of yeast cells under defined photosynthetic conditions. We anticipate that these genetically tractable endosymbiotic platforms can be used for evolutionary studies, particularly related to organelle evolution, and also for synthetic biology applications.

Dissection and enhancement of prebiotic properties of yeast cell wall oligosaccharides through metabolic engineering.

Saccharomyces boulardii is a yeast clinically used for treating various symptoms of gastrointestinal dysbiosis. Despite their genomic relatedness, S. boulardii has a distinctive cell wall oligosaccharide composition compared to baker's yeast S. cerevisiae, such as higher mannan content. Here we explore the beneficial effects of S. boulardii cell wall oligosaccharides through metabolic engineering. We increased the production of guanosine diphosphate (GDP)-mannose, the substrate for cell wall mannan biosynthesis, by perturbing glycolysis flux and overexpressing the enzymes in the GDP-mannose biosynthesis pathway. Combined with overexpression of a cell wall mannoprotein and dolichol phosphate mannose synthase, the cell wall mannan content of S. boulardii increased up to 52%. The identical engineering resulted in marginal changes in the S. cerevisiae cell wall. S. boulardii showed a higher adhesive capacity against Salmonella enterica Typhimurium than S. cerevisiae, and yeast-bacteria sedimentation rates were positively correlated with cell wall mannan contents. Besides, S. boulardii biomass selectively proliferated Bacteroides thetaiotaomicron over Clostridioides difficile more efficiently than S. cerevisiae, and the selectivity was further enhanced by amplifying the cell wall mannan. Collectively, we report the important prebiotic roles of cell wall oligosaccharides in the protective functions of S. boulardii and present a unique metabolic engineering approach to modulate the functions.

PTSD in veterans, couple behavior, and cardiovascular response during marital conflict.

Posttraumatic stress disorder (PTSD) is associated with strain in marriage and similar intimate relationships, and such difficulties could contribute to associations of PTSD with risk of cardiovascular disease (CVD). Heightened cardiovascular reactivity (CVR) during stressful marital interactions may be an important mechanism in this regard. This study examined dysfunctional behavior during marital conflict as a mediator of the association of PTSD with heightened CVR during these interactions. In 64 couples comprising male military veterans and female partners, participants underwent a 17-min video-recorded conflict discussion, with assessment of blood pressure and cardiac sympathetic activation (i.e. preejection period). In half of the couples, veterans met interview and questionnaire criteria for PTSD. Behavior was coded for aspects of affiliation (e.g., warmth vs. hostility) and control (e.g., dominance vs. deference), as well as blends of these broader dimensions. Extending previous reports from this study, actor-partner mediational analyses indicated that PTSD contributed to larger increases in veterans' and spouses' systolic blood pressure during the interaction through effects of the individual's own expressions of low warmth. PTSD contributed to veterans' and spouses' greater cardiac sympathetic activation through effects of the individual's own expressions of hostile control (e.g., blame, criticism). Hence, expressions of low warmth and high hostile control contribute to effects of PTSD on veterans' and spouses' heightened CVR during marital conflict discussions, suggesting a mechanism linking PTSD with CVD risk and potential targets for risk-reducing behavioral interventions. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Resting high frequency heart rate variability and PTSD symptomatology in Veterans: Effects of respiration, role in elevated heart rate, and extension to spouses.

Heart rate variability (HRV) associated with parasympathetic activity (i.e., cardiac vagal tone) is reduced in posttraumatic stress disorder (PTSD), but possible confounding effects of respiration have not been studied sufficiently. Further, reduced parasympathetic inhibition might contribute to elevated heart rate (HR) in PTSD. Finally, reduced HRV in PTSD might extend to intimate partners, given their chronic stress exposure. In 65 couples (male Veterans, female partners), elevated PTSD symptomatology (n = 32; 28 met full DSM IV criteria, 4 fell slightly short) was documented by structured interview and self-reports. Baseline HR, high-frequency HRV (HF-HRV), cardiac pre-ejection period (PEP), and respiration rate and depth were measured via impedance cardiography. Veterans with PTSD symptoms displayed reduced lnHF-HRV, even when adjusting for respiration, but their partners did not. In mediational analyses, elevated resting HR in PTSD was accounted for by lnHF-HRV but not PEP. Results strengthen evidence regarding HF-HRV and elevated HR in PTSD.

Standardized Assessment of Relationship Functioning in Iraq and Afghanistan Veterans with PTSD.

Posttraumatic Stress Disorder (PTSD) is associated with difficulties in intimate relationships, with most prior research examining associations with continuous, single-dimension, and often-unstandardized measures of general relationship quality or aggression. Standardized, well-normed assessments that include multiple couple problem areas could provide more precise information about the presence and specific nature of clinically significant concerns in patient care settings. This investigation aimed to replicate findings regarding increased difficulties in relationship functioning among Operations Enduring and Iraqi Freedom Veterans with PTSD and their romantic partners, specifically using a standardized assessment that permits identification of cases of clinically significant general couple distress and difficulties across multiple problem areas. We compared 32 male Veterans with PTSD and 33 without PTSD, and their romantic partners on reports of several problem areas using the revised Marital Satisfaction Inventory (MSI-R). All participants underwent structured diagnostic interviewing. PTSD couples reported clinically significant levels of relationship distress several times more frequently than comparison couples, both for general distress and across all specific problem areas (e.g., aggressive behavior, quality of leisure time together, sexual functioning, conflicts about finances and child rearing). The most notable problem areas for PTSD couples were affective and problem-solving communication. These results replicate associations of PTSD with general couple discord and multiple specific areas of couple difficulties and extend them by documenting the clinical severity of these problems. Mental health providers may consider incorporating standardized couple assessments into their evaluations of Veterans' functioning. Couples therapies may consider using such measures to prioritize targets for treatment.

Automated Text Message Reminders Improve Radiation Therapy Compliance.

PURPOSE: Protraction of radiation therapy courses can lead to lower cancer control and cancer-specific survival rates. The requirement for daily, consecutive radiation treatments coupled with the complexities of multimodality cancer care and quality assurance can occasionally lead to missed patient appointments or clinical inefficiency. To determine whether an automated text messaging (short message service [SMs]) platform could improve patient compliance with scheduled radiation therapy delivery, we created an automated SMS platform to send daily reminders of radiation therapy appointments. METHODS AND MATERIALS: An automated SMS text messaging program was used from July 2016 to January 2017 to deliver daily appointment time reminders to patients on an elective basis. Automated text messages were sent 2 hours before treatment appointments with appointment-specific information. We analyzed for compliance with radiation therapy appointments for patients who elected to receive SMS reminders versus those who did not. RESULTS: Multivariate analysis of >37,000 encounters involving ∼3400 patients demonstrated that of the factors considered, nonreceipt of SMS appointment reminders had a strong association with 15- to 60-minute tardiness (odds ratio [OR], 1.25; 95% confidence interval [CI], 1.13-1.38; P < .0001), >60-minute tardiness (OR, 1.56; 95% CI, 1.34-1.82; P < .0001) and no-shows (OR, 6.77; 95% CI, 5.45-8.41; P < .0001). Other demographic factors associated with decreased compliance included being early in a radiation therapy course, having an appointment earlier in the day, younger age, and male sex. Receipt of an SMS message did not correlate with overall treatment package time. CONCLUSIONS: Receipt of text messages correlates with compliance for radiation therapy appointments. Prospective randomized trials would be required to determine conclusively whether SMS is an effective intervention for improving compliance in populations at risk for being late to or missing radiation therapy appointments.

Remodeling of the abdominal epithelial monolayer during the larva-pupa-adult transformation of Manduca.

During metamorphosis of insect epithelial monolayers, cells die, divide, and rearrange. In Drosophila undifferentiated diploid cells destined to form the adult cuticle of each abdominal segment segregate early in development from the surrounding polyploid larval epithelial cells of that segment as eight groups of diploid histoblast cells. The larval polyploid cells are programmed to die and be replaced by divisions and rearrangements of histoblast cells. By contrast, abdominal epithelial cells of Manduca larvae form a monolayer of cells representing different ploidy levels with no definitive segregation of diploid cells destined to form adult structures. These epithelial cells of mixed ploidy levels produce a thick smooth larval cuticle with sparsely distributed sensory bristles. Adult descendants of this larval monolayer produce a thinner cuticle with densely packed scale cells. The transition between these differentiated states of Manduca involves divisions of cells, changes in ploidy levels, and sorting of certain polyploid cells into circular rosette patches to minimize contacts of these polyploid cells with surrounding cells of equal or smaller size. Cells within the rosettes and some surrounding cells are destined to die and be replaced by remaining epithelial cells of uniform size and ploidy at pupa-adult apolysis.

Battling on the Home Front: Posttraumatic Stress Disorder and Conflict Behavior Among Military Couples.

This study evaluated interpersonal behavior differences among male military service members with and without PTSD and their female partners. Couples (N = 64) completed a 17-minute videotaped conflict discussion, and their interaction behavior was coded using the circumplex-based Structural Analysis of Social Behavior model (SASB; Benjamin, 1979, 1987, 2000). Within couples, the behavior of partners was very similar. Compared to military couples without PTSD, couples with PTSD displayed more interpersonal hostility and control. Couples with PTSD also exhibited more sulking, blaming, and controlling behavior, and less affirming and connecting behavior, than couples without PTSD. Results advance our understanding of the relational impacts of PTSD on military service members and their partners, and underscore the value of couple-based interventions for PTSD in the context of relationship distress.

Posttraumatic Stress Disorder Symptom Severity and Relationship Functioning Among Partnered Heterosexual and Lesbian Women Veterans.

PURPOSE: Few studies have examined associations of posttraumatic stress disorder (PTSD) and relationship satisfaction among women Veterans, and no research has explored these factors in lesbian women Veterans, a large subgroup that may have unique concerns. This study examined the link between PTSD and relationship satisfaction in partnered heterosexual and lesbian women Veterans and evaluated potential moderation by sexual orientation, partner support, and conflict. METHODS: Women Veterans (heterosexual n = 260; lesbian n = 128) were recruited nationally to complete a cross-sectional online survey. Multiple linear regression models were used to evaluate moderation, using two- and three-way interactions. RESULTS: Partner support moderated the association between PTSD symptoms and relationship satisfaction to a different degree for heterosexual and lesbian women Veterans, playing a more prominent role in this association for lesbian women. Specifically, for lesbians with low partner support, as PTSD symptoms worsened, relationship satisfaction decreased at a steeper rate than for heterosexual women with low partner support. On the other hand, for lesbians with high partner support, as PTSD symptoms worsened, relationship satisfaction decreased less sharply than for heterosexual women with high partner support. Degree of conflict was highly correlated with relationship satisfaction and also appeared to moderate these relations differently by sexual orientation. CONCLUSION: These findings suggest that women Veterans with PTSD experience impairments in their romantic relationships, which vary by sexual orientation, partner support, and conflict. Partner support and conflict may be important targets in assessment and therapy for women Veterans with PTSD, and especially so for sexual minorities.

Evaluation of a Telehealth Service for COPD and HF patients: Clinical outcome and patients' perceptions.

NHS Croydon Health Services and NHS Croydon Clinical Commissioning Group have been providing telehealth services for the past 36 months. The aim of this study was to measure the impact of telehealth when implemented as a service within a standard patient care pathway. To measure the clinical outcomes, the number of A&E visits and hospital admissions, recorded on the NHS Secondary Uses Service database, were compared before and after the implementation of the telehealth service. The number of all events despite its cause and the number of events related to the patients' diagnosed condition were collected. To elicit patients' perceptions about the telehealth service, a cross sectional survey of patients registered on the triage manager database was used to explore their perceptions, concerns and general satisfaction with the telehealth service via a 4 point likert scale questionnaire. The data of 48 patients were collected and telehealth reduced the number of both A&E and hospital admission due to all causes by 13% (P = 0.42) and 22% (P = 0.048), respectively. When only the events directly related to the patient's diagnosed condition were considered, a reduction by 36% (P = 0.03) and 28% (P = 0.02) was recorded for A&E visits and hospital admission respectively. 27 patients consented to participate in the survey. Overall, patients were very satisfied with telehealth services. Patients agreed that telehealth had improved their health, it was a convenient form of health care delivery for them and they were more involved in the decisions about their care or treatment. In addition, since being on telehealth, patients' confidence in managing their health increased from somewhat confident to confident. Telehealth, when provided as a service within a standard care pathway, seems to decrease hospital admissions and A&E visits. Good patient satisfaction suggests that the current service is accepted and it could be further expanded to include a larger number of patients.

The 'Can't Intubate Can't Oxygenate' scenario in Pediatric Anesthesia: a comparison of different devices for needle cricothyroidotomy.

BACKGROUND: Little evidence exists to guide the management of the 'Can't Intubate, Can't Oxygenate' (CICO) scenario in pediatric anesthesia. OBJECTIVES: To compare two intravenous cannulae for ease of use, success rate and complication rate in needle tracheotomy in a postmortem animal model of the infant airway, and trial a commercially available device using the same model. METHODS: Two experienced proceduralists repeatedly attempted cannula tracheotomy in five postmortem rabbits, alternately using 18-gauge (18G) and 14-gauge (14G) BD Insyte(™) cannulae (BD, Franklin Lakes, NJ, USA). Attempts began at the first tracheal cartilage, with subsequent attempts progressively more caudad. Success was defined as intratracheal cannula placement. In each rabbit, an attempt was then made by each proceduralist to perform a cannula tracheotomy using the Quicktrach Child(™) device (VBM Medizintechnik GmbH, Sulz am Neckar, Germany). RESULTS: The rabbit tracheas were of similar dimensions to a human infant. 60 attempts were made at cannula tracheotomy, yielding a 60% success rate. There was no significant difference in success rate, ease of use, or complication rate between cannulae of different gauge. Successful aspiration was highly predictive (positive predictive value 97%) and both sensitive (89%) and specific (96%) for tracheal cannulation. The posterior tracheal wall was perforated in 42% of tracheal punctures. None of 13 attempts using the Quicktrach Child(™) were successful. CONCLUSION: Cannula tracheotomy in a model comparable to the infant airway is difficult and not without complication. Cannulae of 14- and 18-gauge appear to offer similar performance. Successful aspiration is the key predictor of appropriate cannula placement. The Quicktrach Child was not used successfully in this model. Further work is required to compare possible management strategies for the CICO scenario.

Presence of Plasmodium and Haemoproteus in breeding prothonotary warblers (Protonotaria citrea: Parulidae): temporal and spatial trends in infection prevalence.

Prothonotary warbler (Protonotaria citrea) has shown a long-term decline in abundance in the United States. As a long-range migrant, these warblers are exposed to parasites in both tropical and temperate regions. The focus of this study was to use molecular techniques to examine the temporal prevalence patterns of heamosopridian parasites Plasmodium and Haemoproteus in breeding prothonotary warblers. The prevalence (presence or absence) of Plasmodium and Haemoproteus species was assayed using primer sets for the cytochrome b gene of the mitochondrial DNA. Blood samples were obtained from 187 adult prothonotary warblers collected at 3 central Virginia, U.S.A., breeding sites. The relationship between haemosporidian parasite infections and reproductive success also was examined. We found that 71% of captured prothonotary warblers were infected with haemosporidian parasites, specifically, with 36% prevalence for Haemoproteus spp. and 44% prevalence for Plasmodium spp., during the 2008 breeding season; for both parasites, prevalence increased throughout the season. We found significant variation in haemosporidian parasite prevalence across the breeding season that was strongly site specific. Conversely, we found no significant effects of haemosporidian parasite infections on the reproductive success of prothonotary warblers. This is in sharp contrast to recent reports suggesting considerable effects of these parasites on the reproductive success of wild birds.