ARCAPA in Pulmonary Atresia, Ventricular Septal Defect, and Major Aortopulmonary Collateral Arteries.

We report a case of anomalous origin of the right coronary artery from the pulmonary artery in a patient with pulmonary atresia, ventricular septal defect, and major aortopulmonary collateral arteries. The diagnosis was made during a proposed hybrid approach to stent the native right ventricular outflow tract, and an alternative surgical strategy was created.

α-Synuclein expression in response to bacterial ligands and metabolites in gut enteroendocrine cells: an in vitro proof of concept study.

Caudo-rostral migration of pathological forms of α-synuclein from the gut to the brain is proposed as an early feature in Parkinson's disease pathogenesis, but the underlying mechanisms remain unknown. Intestinal epithelial enteroendocrine cells sense and respond to numerous luminal signals, including bacterial factors, and transmit this information to the brain via the enteric nervous system and vagus nerve. There is evidence that gut bacteria composition and their metabolites change in Parkinson's disease patients, and these alterations can trigger α-synuclein pathology in animal models of the disorder. Here, we investigated the effect of toll-like receptor and free fatty acid receptor agonists on the intracellular level of α-synuclein and its release using mouse secretin tumour cell line 1 enteroendocrine cells. Secretin tumour cell line 1 enteroendocrine cells were treated for 24 or 48 h with toll-like receptor agonists (toll-like receptor 4 selective lipopolysaccharide; toll-like receptor 2 selective Pam3CysSerLys4) and the free fatty acid receptor 2/3 agonists butyrate, propionate and acetate. The effect of selective receptor antagonists on the agonists' effects after 24 hours was also investigated. The level of α-synuclein protein was measured in cell lysates and cell culture media by western blot and enzyme-linked immunosorbent assay. The level of α-synuclein and tumour necrosis factor messenger RNA was measured by quantitative reverse transcription real-time polymerase chain reaction. Stimulation of secretin tumour cell line 1 enteroendocrine cells for 24 and 48 hours with toll-like receptor and free fatty acid receptor agonists significantly increased the amount of intracellular α-synuclein and the release of α-synuclein from the cells into the culture medium. Both effects were significantly reduced by antagonists selective for each receptor. Toll-like receptor and free fatty acid receptor agonists also significantly increased tumour necrosis factor transcription, and this was effectively inhibited by corresponding antagonists. Elevated intracellular α-synuclein increases the likelihood of aggregation and conversion to toxic forms. Factors derived from bacteria induce α-synuclein accumulation in secretin tumour cell line 1 enteroendocrine cells. Here, we provide support for a mechanism by which exposure of enteroendocrine cells to specific bacterial factors found in Parkinson's disease gut dysbiosis might facilitate accumulation of α-synuclein pathology in the gut.

A brief transdiagnostic group (the take control course) compared to individual low-intensity CBT for depression and anxiety: a randomized non-inferiority trial.

Few studies have examined brief transdiagnostic groups. The Take Control Course (TCC) was developed for patients with mild to moderate common mental health problems. We examined whether TCC is non-inferior to individual low-intensity cognitive behaviour therapy (CBT) in a single-blind individually randomised parallel non-inferiority trial. The primary outcomes were depression (PHQ9) and anxiety (GAD7) at 6-month follow-up (primary outcome point) and 12-month follow-up. The non-inferiority margin that we set, based on previous trials, corresponds to approximately 3 points on the PHQ9 and approximately 2.5 points on the GAD7. Intention-to-treat (ITT) and per-protocol (PP) analyses of 6-month data of 156 randomised patients indicated that TCC was non-inferior to individual low-intensity CBT on anxiety (ITT Coefficient = 0.24; 95% CI: -1.45 to 1.92; d = 0.04; p = .79), and depression (ITT Coefficient = 0.82; 95% CI: -1.06 to 2.69; d = 0.14; p = .39) outcomes, and functioning (ITT Coefficient = 0.69; 95% CI: -2.56 to 3.94; d = 0.08; p = .68). The findings at 12 months were inconclusive and require further testing. This randomised trial provides preliminary support that TCC is not less effective than short-term individual CBT within Improving Access to Psychological Therapies (IAPT) services.

'Out of sync': a qualitative investigation of patients' experiences of atrial fibrillation and perceptions of weight management.

OBJECTIVES: To explore ways to enhance the design of risk factor management and weight-loss services for people with overweight/obesity and atrial fibrillation (AF). BACKGROUND: AF is the most common cardiac arrhythmia, with serious consequences for health and quality of life. Some evidence indicates weight reduction in people with AF and overweight/obesity may improve symptoms. This population may require additional support with weight management due to factors associated with ageing and health. DESIGN: Qualitative investigation based on semi-structured interviews. METHODS: 12 adult participants (4 female, 8 male) with diagnosed AF and a current or previous body mass index >27 kg/m2 were recruited at a large tertiary cardiac referral centre in southern England between September 2020 and January 2021. Participants completed quality of life and AF symptom questionnaires using Think-Aloud technique and semi-structured interviews relating to their weight management experiences, needs and preferences. Interviews were audio recorded and analysed thematically using the Capability, Opportunity and Motivation-Behaviour model as a theoretical framework. RESULTS: Three main themes were identified. Being out of rhythm explores the psychological and physical impact of AF on weight management; doing the right thing discusses participants' weight management experiences and broaching the subject explores participants' perspectives on weight management conversations with clinicians. CONCLUSIONS: There was dissatisfaction with the weight management advice received from healthcare professionals including cardiologists. Participants wanted open, non-judgemental discussion of cardiac health implications of overweight/obesity supported by referral to weight management services. Improved communication including research findings regarding the benefits of weight loss as a factor in AF management might increase motivation to adhere to weight-loss advice in this population.

Bile acids and neurological disease.

This review will focus on how bile acids are being used in clinical trials to treat neurological diseases due to their central involvement with the gut-liver-brain axis and their physiological and pathophysiological roles in both normal brain function and multiple neurological diseases. The synthesis of primary and secondary bile acids species and how the regulation of the bile acid pool may differ between the gut and brain is discussed. The expression of several bile acid receptors in brain and their currently known functions along with the tools available to manipulate them pharmacologically are examined, together with discussion of the interaction of bile acids with the gut microbiome and their lesser-known effects upon brain glucose and lipid metabolism. How dysregulation of the gut microbiome, aging and sex differences may lead to disruption of bile acid signalling and possible causal roles in a number of neurological disorders are also considered. Finally, we discuss how pharmacological treatments targeting bile acid receptors are currently being tested in an array of clinical trials for several different neurodegenerative diseases.

Lymphotoxin-alpha expression in the meninges causes lymphoid tissue formation and neurodegeneration.

Organized meningeal immune cell infiltrates are suggested to play an important role in cortical grey matter pathology in the multiple sclerosis brain, but the mechanisms involved are as yet unresolved. Lymphotoxin-alpha plays a key role in lymphoid organ development and cellular cytotoxicity in the immune system and its expression is increased in the CSF of naïve and progressive multiple sclerosis patients and post-mortem meningeal tissue. Here we show that persistently increased levels of lymphotoxin-alpha in the cerebral meninges can give rise to lymphoid-like structures and underlying multiple sclerosis-like cortical pathology. Stereotaxic injections of recombinant lymphotoxin-alpha into the rat meninges led to acute meningeal inflammation and subpial demyelination that resolved after 28 days, with demyelination being dependent on prior subclinical immunization with myelin oligodendrocyte glycoprotein. Injection of a lymphotoxin-alpha lentiviral vector into the cortical meningeal space, to produce chronic localized overexpression of the cytokine, induced extensive lymphoid-like immune cell aggregates, maintained over 3 months, including T-cell rich zones containing podoplanin + fibroblastic reticular stromal cells and B-cell rich zones with a network of follicular dendritic cells, together with expression of lymphoid chemokines and their receptors. Extensive microglial and astroglial activation, subpial demyelination and marked neuronal loss occurred in the underlying cortical parenchyma. Whereas subpial demyelination was partially dependent on previous myelin oligodendrocyte glycoprotein immunization, the neuronal loss was present irrespective of immunization. Conditioned medium from LTα treated microglia was able to induce a reactive phenotype in astrocytes. Our results show that chronic lymphotoxin-alpha overexpression alone is sufficient to induce formation of meningeal lymphoid-like structures and subsequent neurodegeneration, similar to that seen in the progressive multiple sclerosis brain.

The development, validation and normative data study of the English in Ireland adaption of the Philadelphia repeatable Verbal Learning Test (EirPrVLT-12) for use in an older adult population.

OBJECTIVE: Cultural adaptations of verbal serial list-learning tests such as the California Verbal Learning Test (CVLT) and the Philadelphia (repeatable) Verbal Learning Test (P(r)VLT) have been shown to be clinically necessary. This paper aimed to culturally adapt, validate and provide normative data for an English in Ireland adaptation of the P(r)VLT, i.e. the EirPrVLT-12, in order to improve episodic memory assessments for Irish adults. Method: EirPrVLT-12 word lists were constructed using a word frequency study of Irish adults (n = 58). Two twelve-word, four-trial forms were constructed (standard and alternate form). A normative study included 145 participants who met strict inclusion criteria. Results: EirPrVLT-12 performance varied depending on age, gender, education, estimated IQ and socioeconomic status. Construct validity was established by correlations with other cognitive tests. Principal component analysis yielded a three-factor solution relating to general verbal learning, intrusions and interference. Normed EirPrVLT-12 scaled scores and percentiles stratified by age are available on the Open Science Framework at https://osf.io/vjzsp/, as are regression equations to predict individual scores based on age, gender and education. Conclusions: The data obtained underscores the clinical ultility of the EirPrVLT-12 to assess episodic memory in Irish older adults. Future research was recommended to validate the EirPrVLT-12 in a clinical population, extend normative data to younger populations and develop norms for the alternate form.

Heterogeneity in human hippocampal CaMKII transcripts reveals allosteric hub-dependent regulation.

Calcium/calmodulin-dependent protein kinase II (CaMKII) plays a central role in Ca2+ signaling throughout the body. In the hippocampus, CaMKII is required for learning and memory. Vertebrate genomes encode four CaMKII homologs: CaMKIIα, CaMKIIß, CaMKIIγ, and CaMKIIδ. All CaMKIIs consist of a kinase domain, a regulatory segment, a variable linker region, and a hub domain, which is responsible for oligomerization. The four proteins differ primarily in linker length and composition because of extensive alternative splicing. Here, we report the heterogeneity of CaMKII transcripts in three complex samples of human hippocampus using deep sequencing. We showed that hippocampal cells contain a diverse collection of over 70 CaMKII transcripts from all four CaMKII-encoding genes. We characterized the Ca2+/CaM sensitivity of hippocampal CaMKII variants spanning a broad range of linker lengths and compositions. The effect of the variable linker on Ca2+/CaM sensitivity depended on the kinase and hub domains. Moreover, we revealed a previously uncharacterized role for the hub domain as an allosteric regulator of kinase activity, which may provide a pharmacological target for modulating CaMKII activity. Using small-angle x-ray scattering and single-particle cryo-electron microscopy (cryo-EM), we present evidence for extensive interactions between the kinase and the hub domains, even in the presence of a 30-residue linker. Together, these data suggest that Ca2+/CaM sensitivity in CaMKII is homolog dependent and includes substantial contributions from the hub domain. Our sequencing approach, combined with biochemistry, provides insights into understanding the complex pool of endogenous CaMKII splice variants.

Characterization of the Streptomyces coelicolor Glycoproteome Reveals Glycoproteins Important for Cell Wall Biogenesis.

The physiological role of protein O-glycosylation in prokaryotes is poorly understood due to our limited knowledge of the extent of their glycoproteomes. In Actinobacteria, defects in protein O-mannosyl transferase (Pmt)-mediated protein O-glycosylation have been shown to significantly retard growth (Mycobacterium tuberculosis and Corynebacterium glutamicum) or result in increased sensitivities to cell wall-targeting antibiotics (Streptomyces coelicolor), suggesting that protein O-glycosylation has an important role in cell physiology. Only a single glycoprotein (SCO4142, or PstS) has been identified to date in S. coelicolor Combining biochemical and mass spectrometry-based approaches, we have isolated and characterized the membrane glycoproteome in S. coelicolor A total of ninety-five high-confidence glycopeptides were identified which mapped to thirty-seven new S. coelicolor glycoproteins and a deeper understanding of glycosylation sites in PstS. Glycosylation sites were found to be modified with up to three hexose residues, consistent with what has been observed previously in other ActinobacteriaS. coelicolor glycoproteins have diverse roles and functions, including solute binding, polysaccharide hydrolases, ABC transporters, and cell wall biosynthesis, the latter being of potential relevance to the antibiotic-sensitive phenotype of pmt mutants. Null mutants in genes encoding a putative d-Ala-d-Ala carboxypeptidase (SCO4847) and an l,d-transpeptidase (SCO4934) were hypersensitive to cell wall-targeting antibiotics. Additionally, the sco4847 mutants displayed an increased susceptibility to lysozyme treatment. These findings strongly suggest that both glycoproteins are required for maintaining cell wall integrity and that glycosylation could be affecting enzyme function.IMPORTANCE In prokaryotes, the role of protein glycosylation is poorly understood due to our limited understanding of their glycoproteomes. In some Actinobacteria, defects in protein O-glycosylation have been shown to retard growth and result in hypersensitivity to cell wall-targeting antibiotics, suggesting that this modification is important for maintaining cell wall structure. Here, we have characterized the glycoproteome in Streptomyces coelicolor and shown that glycoproteins have diverse roles, including those related to solute binding, ABC transporters, and cell wall biosynthesis. We have generated mutants encoding two putative cell wall-active glycoproteins and shown them to be hypersensitive to cell wall-targeting antibiotics. These findings strongly suggest that both glycoproteins are required for maintaining cell wall integrity and that glycosylation affects enzyme function.

Uncovering the molecular mechanisms of lignocellulose digestion in shipworms.

Lignocellulose forms the structural framework of woody plant biomass and represents the most abundant carbon source in the biosphere. Turnover of woody biomass is a critical component of the global carbon cycle, and the enzymes involved are of increasing industrial importance as industry moves away from fossil fuels to renewable carbon resources. Shipworms are marine bivalve molluscs that digest wood and play a key role in global carbon cycling by processing plant biomass in the oceans. Previous studies suggest that wood digestion in shipworms is dominated by enzymes produced by endosymbiotic bacteria found in the animal's gills, while little is known about the identity and function of endogenous enzymes produced by shipworms. Using a combination of meta-transcriptomic, proteomic, imaging and biochemical analyses, we reveal a complex digestive system dominated by uncharacterized enzymes that are secreted by a specialized digestive gland and that accumulate in the cecum, where wood digestion occurs. Using a combination of transcriptomics, proteomics, and microscopy, we show that the digestive proteome of the shipworm Lyrodus pedicellatus is mostly composed of enzymes produced by the animal itself, with a small but significant contribution from symbiotic bacteria. The digestive proteome is dominated by a novel 300 kDa multi-domain glycoside hydrolase that functions in the hydrolysis of ß-1,4-glucans, the most abundant polymers in wood. These studies allow an unprecedented level of insight into an unusual and ecologically important process for wood recycling in the marine environment, and open up new biotechnological opportunities in the mobilization of sugars from lignocellulosic biomass.

An ancient family of lytic polysaccharide monooxygenases with roles in arthropod development and biomass digestion.

Thermobia domestica belongs to an ancient group of insects and has a remarkable ability to digest crystalline cellulose without microbial assistance. By investigating the digestive proteome of Thermobia, we have identified over 20 members of an uncharacterized family of lytic polysaccharide monooxygenases (LPMOs). We show that this LPMO family spans across several clades of the Tree of Life, is of ancient origin, and was recruited by early arthropods with possible roles in remodeling endogenous chitin scaffolds during development and metamorphosis. Based on our in-depth characterization of Thermobia's LPMOs, we propose that diversification of these enzymes toward cellulose digestion might have endowed ancestral insects with an effective biochemical apparatus for biomass degradation, allowing the early colonization of land during the Paleozoic Era. The vital role of LPMOs in modern agricultural pests and disease vectors offers new opportunities to help tackle global challenges in food security and the control of infectious diseases.

Prospective cohort feasibility study of a transdiagnostic group intervention for common mental health problems: The Take Control Course.

OBJECTIVES: While transdiagnostic psychological treatments appear to be promising, they require greater empirical support. Further, a number of available transdiagnostic treatments are targeted at clients with a specific category of disorder, such as clients with anxiety disorders. This study is a preliminary examination of the effectiveness, feasibility, and acceptability of a new transdiagnostic six-session group-based intervention (Take Control Course; TCC) predominantly aimed at clients within primary care. The group is aimed at a broad range of clients; it is derived from an integrative transdiagnostic theory, which specifies mechanisms of psychopathology across disorders. Briefer interventions are gaining an increasing evidence base, and this study seeks to compare the TCC to an established brief intervention within primary care. DESIGN: Prospective cohort study comparing two active psychological treatments. METHODS: Take Control Course group (n = 66) was compared to a non-randomized control group of clients accessing individual low-intensity interventions (n = 43) using random-effect regression models. Primary outcomes were depression and anxiety scores; additional outcomes included social and other functioning. RESULTS: For the TCC group, changes on all pre-post outcomes were significant with moderate effect sizes. The between-group differences were not significant. CONCLUSIONS: Results show potential for TCC to be an effective intervention, but further work is required to validate these findings in a more rigorous, randomized study. PRACTITIONER POINTS: Transdiagnostic understandings of psychological distress may inform pragmatic and effective treatments that can be offered to a broad range of clients. This study describes a transdiagnostic intervention (TCC) that targets maintenance processes common across disorders, and presents initial outcome data. The TCC was found to reduce pre-post scores on measures of anxiety and depression.

Plant science. Morphinan biosynthesis in opium poppy requires a P450-oxidoreductase fusion protein.

Morphinan alkaloids from the opium poppy are used for pain relief. The direction of metabolites to morphinan biosynthesis requires isomerization of (S)- to (R)-reticuline. Characterization of high-reticuline poppy mutants revealed a genetic locus, designated STORR [(S)- to (R)-reticuline] that encodes both cytochrome P450 and oxidoreductase modules, the latter belonging to the aldo-keto reductase family. Metabolite analysis of mutant alleles and heterologous expression demonstrate that the P450 module is responsible for the conversion of (S)-reticuline to 1,2-dehydroreticuline, whereas the oxidoreductase module converts 1,2-dehydroreticuline to (R)-reticuline rather than functioning as a P450 redox partner. Proteomic analysis confirmed that these two modules are contained on a single polypeptide in vivo. This modular assembly implies a selection pressure favoring substrate channeling. The fusion protein STORR may enable microbial-based morphinan production.

Metastatic prostate cancer mimicking chronic subdural hematoma: a case report and review of the literature.

Cancer of the prostate is extremely common and is well known to metastasize to the pelvic lymph nodes and axial skeleton (vertebral column, pelvis, cranium, and proximal femur). However, reports of intracranial metastasis are rare and commonly discovered postmortem. Moreover, metastatic lesions mimicking subdural hematoma are extremely rare and are uncommonly reported in the literature. We found only three such cases in the literature. We present a unique case of metastatic prostate cancer presenting with headaches after head trauma with classic radiologic findings of subdural hematoma. The diagnosis may have been made sooner with preoperative magnetic resonance imaging.

Validation of multiplex microbead immunoassay for simultaneous serodetection of multiple infectious agents in laboratory mouse.

Multiplex methodologies enable simultaneous detection of antibodies against several infectious agents allowing sample conservation, cost effectiveness, and amenability to high-throughput/automation. We have previously described a multiplex microbead immunoassay for serodetection of ten, high-priority mouse infectious pathogens. Here, we present a validation of this multiplex diagnostic system using approximately four hundred serum samples from different groups of mice. Computer assisted multivariate analysis of the resulting high volume data (8000 data points) was performed. This computational approach enabled presentation of data in a variety of easily interpretable formats (e.g., correlation tables and heat maps). Importantly, this computer aided approach was instrumental for the evaluation of assay accuracy, sensitivity, specificity, and robustness during the study. Crucial pieces of information were obtained to make timely adjustments for assay refinement. This progressive approach to developing an implementation-ready clinical assay, facilitated by computational analysis, produced a highly efficient, accurate and dependable serodiagnostics system. This system has effectively replaced the current state-of-the-art methodology (ELISA) used in mouse colony health management at the University of California and the Jackson Laboratory. A pathway to develop multiplex serology tests for infectious disease diagnosis described here serves as a model for multiplex immunoassay design, clinical validation, refinement and implementation.

The relation of post-traumatic stress symptoms to depression and pain in patients with accident-related chronic pain.

UNLABELLED: Symptoms of post-traumatic stress disorder (PTSD) are a common comorbidity in patients with a history of accident-related chronic pain and depression. However, little is known regarding the influence of PTSD in contributing to the affective distress, pain experience, and disability associated with chronic pain in this population. This study used structural equation modeling to examine 3 models that assess these relations in a sample of chronic pain patients with accident-related pain. Subjects were assessed for pain experience, depressive symptoms, anxiety, PTSD symptoms, pain disability, and relevant demographic variables. Pearson correlations indicated that symptoms of depression were significantly related to more severe pain, disability, and PTSD symptoms. PTSD symptoms were significantly associated with higher disability. The model of best fit indicated that after controlling for the influence of anxiety on the dependent measures, PTSD symptoms have a direct influence on severity of depressive symptoms, whereas depressive symptoms have a direct influence on pain intensity and an indirect impact on pain intensity by way of their effect on disability. These data point to the importance of unresolved PTSD symptoms in contributing to the level of depression, pain, and disability exhibited by chronic pain patients and highlight the need to consider directed and primary treatment of PTSD in pain rehabilitation programs. PERSPECTIVE: This study highlights the impact of symptoms of PTSD on levels of depression, disability, and pain in patients with pain secondary to physical injury. Our results suggest that pain rehabilitation programs provide directed interventions for PTSD symptoms among this population to improve pain treatment outcomes.

Assessing the risk of transmission of three infectious agents among mice housed in a negatively pressurized caging system.

Previous studies from our institution have shown that ventilated caging run at negative pressure to a mouse room dramatically reduced exposure of personnel to the major mouse allergen, Mus m 1. The current study was designed to determine whether negative cage ventilation posed an inordinate risk for spread of infectious agents between cages and/or racks. B6;129S-Tnfsf5(tm1Imx)/J (TNF) mice, which were naturally and persistently infected with Pneumocystis carinii, Helicobacter bilis, and Pasteurella pneumotropica, were used as the source of infections. Uninfected C3Smn.CB17-Prkdc(scid)/J (SCID) mice with severe combined immunodeficiency were used to detect transmission. The following methods were used to detect transmission of infections: polymerase chain reaction (PCR) amplification and histological examination of lungs for P. carinii; PCR of fecal specimens or cecal contents for H. bilis; and culture of oropharyngeal, tracheal, or vaginal swabs for P. pneumotropica. We determined whether transmission of the three agents occurred via direct contact (cohabitation), exposure to soiled bedding, and/or by handling naive SCID mice after handling infected TNF mice. During a 12-week period, all three infectious agents were readily transmitted to uninfected mice by cohabitation. Transmission was much less efficient and occurred later among mice exposed to contaminated bedding. Transmission did not occur as a result of handling. We then studied transmission of the three infectious agents among mice housed in individually ventilated cages run at negative pressure in a small, crowded mouse room. Transmission of P. carinii was detected at the end of the 12-month study in the densely populated room, probably because the exhaust from the changing station passed over soiled cages and caused aerosolization of particulates. Caging systems run at negative pressure effectively reduce personnel exposure to allergens and may also inhibit the transmission of infectious diseases.