Stress Inoculation Training (SIT-NORCAL), Part 1: The Development and Preliminary Evaluation of a Psychological Performance Training Protocol.

BACKGROUND: Stress inoculation training (SIT) interventions have demonstrated promise within military contexts for human performance enhancement and psychological health applications. However, lack of manualized guidance on core content selection, delivery, and measurement processes has limited their use. PURPOSE: The purpose of this study was to develop and evaluate a comprehensive SIT intervention protocol to enhance the performance and health of military personnel engaged in special warfare and first-response activities. METHODS: Multidisciplinary teams of subject matter experts (n = 19) were consulted in protocol generation. The performance improvement/human performance technology (HPT) model was used in the selection, refinement, and measurement of core skills. The protocol was trialed and refined (44 cohorts, n = =300; 2013-2020) to generate the results. RESULTS: Four primary aims were achieved: (1) The generation of a flexible, evidence-based/evidence-driven psychological performance and health sustainment hybrid, SIT-NORCAL. (2) Manualized content and process guidance. (3) The creation of multimedia materials using evidence-based methodologies. (4) The design of initial measurement systems. Preliminary quality improvement analysis demonstrated positive results using standard-of-care and performance enhancement assessments. CONCLUSION: Hybridized human performance and psychological health sustainment protocols represent a paradigm shift in the delivery of psychological performance training with the potential to overcome barriers to success in traditional care. Further study is needed to determine the effectiveness and reach of SIT-NORCAL.

Stress Inoculation Training (SIT-NORCAL), Part 2: A Pilot Study Among Explosive Ordnance Disposal Special Warfare Enablers.

BACKGROUND: Despite being a well-supported strategy, Stress Inoculation Training (SIT) has not been fully incorporated in the advancement of human performance among most military personnel. The RAND Study recommendations for maximizing SIT's potential within high-risk/ high-intensity occupational groups were used in designing the Core Training protocol targeting psychological performance, SIT-NORCAL (Part 1). PURPOSE: The current project (Part 2) sought to further develop the protocol as a health and human performance hybrid through quality improvement analysis of the content, process, and measurement elements for use in the human performance context. METHODS: Evidence-based/evidence-driven methodologies were used in collaborative design tailored to the unique needs of special warfare enablers specializing in Explosive Ordnance Disposal (n = 17). The resultant three-phase training was conducted with a novice group (n = 10) using standardized measurements of collaboration, human performance, and adaptive capabilities on identified training targets. RESULTS: Process elements demonstrated high feasibility, resulting in high collaboration and trainee satisfaction. Significant improvements in psychological performance targets were observed pre- to post-training, and during an Adaptive Environmental Simulation designed by unit members. Two weeks post-training, unit members (n = 5) responded to an actual crash of an F-16 aircraft; measurements indicated maintenance of skill set from training to real-world events. CONCLUSION: Deployment of the elements in the SIT-NORCAL protocol demonstrated early feasibility and positive training impact on occupationally relevant skills that carried over into real-world events.

The Preservative Sorbic Acid Targets Respiration, Explaining the Resistance of Fermentative Spoilage Yeast Species.

A small number (10 to 20) of yeast species cause major spoilage in foods. Spoilage yeasts of soft drinks are resistant to preservatives like sorbic acid, and they are highly fermentative, generating large amounts of carbon dioxide gas. Conversely, many yeast species derive energy from respiration only, and most of these are sorbic acid sensitive and so prevented from causing spoilage. This led us to hypothesize that sorbic acid may specifically inhibit respiration. Tests with respirofermentative yeasts showed that sorbic acid was more inhibitory to both Saccharomyces cerevisiae and Zygosaccharomyces bailii during respiration (of glycerol) than during fermentation (of glucose). The respiration-only species Rhodotorula glutinis was equally sensitive when growing on either carbon source, suggesting that ability to ferment glucose specifically enables sorbic acid-resistant growth. Sorbic acid inhibited the respiration process more strongly than fermentation. We present a data set supporting a correlation between the level of fermentation and sorbic acid resistance across 191 yeast species. Other weak acids, C2 to C8, inhibited respiration in accordance with their partition coefficients, suggesting that effects on mitochondrial respiration were related to membrane localization rather than cytosolic acidification. Supporting this, we present evidence that sorbic acid causes production of reactive oxygen species, the formation of petite (mitochondrion-defective) cells, and Fe-S cluster defects. This work rationalizes why yeasts that can grow in sorbic acid-preserved foods tend to be fermentative in nature. This may inform more-targeted approaches for tackling these spoilage organisms, particularly as the industry migrates to lower-sugar drinks, which could favor respiration over fermentation in many spoilage yeasts.IMPORTANCE Spoilage by yeasts and molds is a major contributor to food and drink waste, which undermines food security. Weak acid preservatives like sorbic acid help to stop spoilage, but some yeasts, commonly associated with spoilage, are resistant to sorbic acid. Different yeasts generate energy for growth by the processes of respiration and/or fermentation. Here, we show that sorbic acid targets the process of respiration, so fermenting yeasts are more resistant. Fermentative yeasts are also those usually found in spoilage incidents. This insight helps to explain the spoilage of sorbic acid-preserved foods by yeasts and can inform new strategies for effective control. This is timely as the sugar content of products like soft drinks is being lowered, which may favor respiration over fermentation in key spoilage yeasts.

Method for RNA extraction and transcriptomic analysis of single fungal spores.

Transcriptomic analysis of single cells has been increasingly in demand in recent years, thanks to technological and methodological advances as well as growing recognition of the importance of individuals in biological systems. However, the majority of these studies have been performed in mammalian cells, due to their ease of lysis and high RNA content. No single cell transcriptomic analysis has yet been described in microbial spores, even though it is known that heterogeneity at the phenotype level exists among individual spores. Transcriptomic analysis of single spores is challenging, in part due to the physically robust nature of the spore wall. This precludes the use of methods commonly used for mammalian cells. Here, we describe a simple method for extraction and amplification of transcripts from single fungal conidia (asexual spores), and its application in single-cell transcriptomics studies. The method can also be used for studies of small numbers of fungal conidia, which may be necessary in the case of limited sample availability, low-abundance transcripts or interest in small subpopulations of conidia. •The method allows detection of transcripts from single conidia of Aspergillus niger•The method allows detection of genomic DNA from single conidia of Aspergillus niger.

Weak Acid Resistance A (WarA), a Novel Transcription Factor Required for Regulation of Weak-Acid Resistance and Spore-Spore Heterogeneity in Aspergillus niger.

Propionic, sorbic, and benzoic acids are organic weak acids that are widely used as food preservatives, where they play a critical role in preventing microbial growth. In this study, we uncovered new mechanisms of weak-acid resistance in molds. By screening a library of 401 transcription factor deletion strains in Aspergillus fumigatus for sorbic acid hypersensitivity, a previously uncharacterized transcription factor was identified and named weak acid resistance A (WarA). The orthologous gene in the spoilage mold Aspergillus niger was identified and deleted. WarA was required for resistance to a range of weak acids, including sorbic, propionic, and benzoic acids. A transcriptomic analysis was performed to characterize genes regulated by WarA during sorbic acid treatment in A. niger Several genes were significantly upregulated in the wild type compared with a ΔwarA mutant, including genes encoding putative weak-acid detoxification enzymes and transporter proteins. Among these was An14g03570, a putative ABC-type transporter which we found to be required for weak-acid resistance in A. niger We also show that An14g03570 is a functional homologue of the Saccharomyces cerevisiae protein Pdr12p and we therefore name it PdrA. Last, resistance to sorbic acid was found to be highly heterogeneous within genetically uniform populations of ungerminated A. niger conidia, and we demonstrate that pdrA is a determinant of this heteroresistance. This study has identified novel mechanisms of weak-acid resistance in A. niger which could help inform and improve future food spoilage prevention strategies.IMPORTANCE Weak acids are widely used as food preservatives, as they are very effective at preventing the growth of most species of bacteria and fungi. However, some species of molds can survive and grow in the concentrations of weak acid employed in food and drink products, thereby causing spoilage with resultant risks for food security and health. Current knowledge of weak-acid resistance mechanisms in these fungi is limited, especially in comparison to that in yeasts. We characterized gene functions in the spoilage mold species Aspergillus niger which are important for survival and growth in the presence of weak-acid preservatives. Such identification of weak-acid resistance mechanisms in spoilage molds will help in the design of new strategies to reduce food spoilage in the future.

Molecular patterns of response and treatment failure after frontline venetoclax combinations in older patients with AML.

The BCL-2 inhibitor venetoclax combined with hypomethylating agents or low-dose cytarabine represents an important new therapy for older or unfit patients with acute myeloid leukemia (AML). We analyzed 81 patients receiving these venetoclax-based combinations to identify molecular correlates of durable remission, response followed by relapse (adaptive resistance), or refractory disease (primary resistance). High response rates and durable remissions were typically associated with NPM1 or IDH2 mutations, with prolonged molecular remissions prevalent for NPM1 mutations. Primary and adaptive resistance to venetoclax-based combinations was most commonly characterized by acquisition or enrichment of clones activating signaling pathways such as FLT3 or RAS or biallelically perturbing TP53. Single-cell studies highlighted the polyclonal nature of intratumoral resistance mechanisms in some cases. Among cases that were primary refractory, we identified heterogeneous and sometimes divergent interval changes in leukemic clones within a single cycle of therapy, highlighting the dynamic and rapid occurrence of therapeutic selection in AML. In functional studies, FLT3 internal tandem duplication gain or TP53 loss conferred cross-resistance to both venetoclax and cytotoxic-based therapies. Collectively, we highlight molecular determinants of outcome with clinical relevance to patients with AML receiving venetoclax-based combination therapies.

Investigating chitin deacetylation and chitosan hydrolysis during vegetative growth in Magnaporthe oryzae.

Chitin deacetylation results in the formation of chitosan, a polymer of ß1,4-linked glucosamine. Chitosan is known to have important functions in the cell walls of a number of fungal species, but its role during hyphal growth has not yet been investigated. In this study, we have characterized the role of chitin deacetylation during vegetative hyphal growth in the filamentous phytopathogen Magnaporthe oryzae. We found that chitosan localizes to the septa and lateral cell walls of vegetative hyphae and identified 2 chitin deacetylases expressed during vegetative growth-CDA1 and CDA4. Deletion strains and fluorescent protein fusions demonstrated that CDA1 is necessary for chitin deacetylation in the septa and lateral cell walls of mature hyphae in colony interiors, whereas CDA4 deacetylates chitin in the hyphae at colony margins. However, although the Δcda1 strain was more resistant to cell wall hydrolysis, growth and pathogenic development were otherwise unaffected in the deletion strains. The role of chitosan hydrolysis was also investigated. A single gene encoding a putative chitosanase (CSN) was discovered in M. oryzae and found to be expressed during vegetative growth. However, chitosan localization, vegetative growth, and pathogenic development were unaffected in a CSN deletion strain, rendering the role of this enzyme unclear.

Chitosan Mediates Germling Adhesion in Magnaporthe oryzae and Is Required for Surface Sensing and Germling Morphogenesis.

The fungal cell wall not only plays a critical role in maintaining cellular integrity, but also forms the interface between fungi and their environment. The composition of the cell wall can therefore influence the interactions of fungi with their physical and biological environments. Chitin, one of the main polysaccharide components of the wall, can be chemically modified by deacetylation. This reaction is catalyzed by a family of enzymes known as chitin deacetylases (CDAs), and results in the formation of chitosan, a polymer of ß1,4-glucosamine. Chitosan has previously been shown to accumulate in the cell wall of infection structures in phytopathogenic fungi. Here, it has long been hypothesized to act as a 'stealth' molecule, necessary for full pathogenesis. In this study, we used the crop pathogen and model organism Magnaporthe oryzae to test this hypothesis. We first confirmed that chitosan localizes to the germ tube and appressorium, then deleted CDA genes on the basis of their elevated transcript levels during appressorium differentiation. Germlings of the deletion strains showed loss of chitin deacetylation, and were compromised in their ability to adhere and form appressoria on artificial hydrophobic surfaces. Surprisingly, the addition of exogenous chitosan fully restored germling adhesion and appressorium development. Despite the lack of appressorium development on artificial surfaces, pathogenicity was unaffected in the mutant strains. Further analyses demonstrated that cuticular waxes are sufficient to over-ride the requirement for chitosan during appressorium development on the plant surface. Thus, chitosan does not have a role as a 'stealth' molecule, but instead mediates the adhesion of germlings to surfaces, thereby allowing the perception of the physical stimuli necessary to promote appressorium development. This study thus reveals a novel role for chitosan in phytopathogenic fungi, and gives further insight into the mechanisms governing appressorium development in M.oryzae.

Phenylethanoid glycosides in tepals of Magnolia salicifolia and their occurrence in flowers of Magnoliaceae.

Phenylethanoid glycosides were among the major UV-absorbing components in 80% aq. CH3OH extracts of the tepals of Magnolia salicifolia (Siebold & Zucc.) Maxim. (Magnoliaceae; Magnolia subgenus Yulania). Structural characterisation of isolated compounds by spectroscopic and chemical methods revealed three previously unrecorded examples, yulanoside A, yulanoside B and 2'-rhamnoechinacoside, and the known compounds echinacoside and crassifolioside; chromatographic methods also identified verbascoside in the tepal extract. Yulanoside A is the first reported example of a phenylethanoid pentaglycoside, namely hydroxytyrosol 1-O-{ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→6)-[3,4-dihydroxycinnamoyl-(→4)][α-L-rhamnopyranosyl-(1→3)][α-L-rhamnopyranosyl-(1→2)]-ß-D-glucopyranoside}. A survey of Magnolia sensu lato and Liriodendron (the two genera of Magnoliaceae) suggested that yulanoside A and its deglucosyl derivative (yulanoside B) were a feature of the tepal chemistry of Magnolia subgenus Yulania (except Magnolia acuminata, the sole member of section Tulipastrum, which did not accumulate phenylethanoid glycosides). The two species of Liriodendron and examined examples of Magnolia subgenus Magnolia sections Magnolia and Rytidospermum (subsection Oyama) also accumulated phenylethanoid glycosides in their tepals and in these species, and in subgenus Yulania, the major compounds were one or more of echinacoside, 2'-rhamnoechinacoside, crassifolioside and verbascoside. Levels of phenylethanoid glycosides were found to be much lower in species studied from Magnolia sections Gwillimia, Macrophylla and Rytidospermum (subsection Rytidospermum), although yulanoside A was detectable in M. macrophylla and this may have some bearing on the placement of section Macrophylla, which is currently uncertain. In the isolates of yulanoside B and echinacoside, minor phenylethanoid glycosides were determined to be analogues of these compounds with ß-D-xylose at C-3' of the primary glucose rather than α-L-rhamnose.

Temporal response of arterial stiffness to ultra-marathon.

The purpose of this investigation was to characterize the arterial stiffness of male ultra-marathon runners (n = 9) using pulse wave velocity (cfPWV) and radial tonometry over the course of an ultra-marathon and during recovery. Measures were collected at rest, immediately following 45 km/75 km of running, then following 60 and 90 min of recovery. No statistical difference was found between baseline cfPWV and normative values. The cfPWV of ultra-endurance runners decreased at 45 km (3.4 ± 1.6 m/s, p=0.006), followed by an increase (1.6 ± 1.8 m/s, p = 0.04) toward baseline levels at the 75 km mark. Radial tonometry measures also indicated small artery stiffness was transiently increased after 75 km. The amount of training time (r = 0.82, p = 0.007) and the duration of a typical training session (r = 0.73, p = 0.03) were correlated strongly with persisting decrements in large artery compliance at 60 min of recovery. The finding that arterial stiffness decreased at the 45 km distance and then reverted back toward baseline levels with prolonged running, may indicate a role of exercise duration or accumulated stress for affecting vascular compliance. At present, it is premature to suggest that athletes should alter training or racing practices to protect vascular health.

Establishing optimal quantitative-polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F-associated myeloproliferative neoplasms: a joint European LeukemiaNet/MPN&MPNr-EuroNet (COST action BM0902) study.

Reliable detection of JAK2-V617F is critical for accurate diagnosis of myeloproliferative neoplasms (MPNs); in addition, sensitive mutation-specific assays can be applied to monitor disease response. However, there has been no consistent approach to JAK2-V617F detection, with assays varying markedly in performance, affecting clinical utility. Therefore, we established a network of 12 laboratories from seven countries to systematically evaluate nine different DNA-based quantitative PCR (qPCR) assays, including those in widespread clinical use. Seven quality control rounds involving over 21,500 qPCR reactions were undertaken using centrally distributed cell line dilutions and plasmid controls. The two best-performing assays were tested on normal blood samples (n=100) to evaluate assay specificity, followed by analysis of serial samples from 28 patients transplanted for JAK2-V617F-positive disease. The most sensitive assay, which performed consistently across a range of qPCR platforms, predicted outcome following transplant, with the mutant allele detected a median of 22 weeks (range 6-85 weeks) before relapse. Four of seven patients achieved molecular remission following donor lymphocyte infusion, indicative of a graft vs MPN effect. This study has established a robust, reliable assay for sensitive JAK2-V617F detection, suitable for assessing response in clinical trials, predicting outcome and guiding management of patients undergoing allogeneic transplant.

Azacitidine fails to eradicate leukemic stem/progenitor cell populations in patients with acute myeloid leukemia and myelodysplasia.

Epigenetic therapies demonstrate significant clinical activity in acute myeloid leukemia (AML) and myelodysplasia (MDS) and constitute an important new class of therapeutic agents. However hematological responses are not durable and disease relapse appears inevitable. Experimentally, leukemic stem/progenitor cells (LSC) propagate disease in animal models of AML and it has been postulated that their relative chemo-resistance contributes to disease relapse. We serially measured LSC numbers in patients with high-risk AML and MDS treated with 5'-azacitidine and sodium valproate (VAL-AZA). Fifteen out of seventy-nine patients achieved a complete remission (CR) or complete remission with incomplete blood count recovery (CRi) with VAL-AZA therapy. There was no significant reduction in the size of the LSC-containing population in non-responders. While the LSC-containing population was substantially reduced in all patients achieving a CR/CRi it was never eradicated and expansion of this population antedated morphological relapse. Similar studies were performed in seven patients with newly diagnosed AML treated with induction chemotherapy. Eradication of the LSC-containing population was observed in three patients all of whom achieved a durable CR in contrast to patients with resistant disease where LSC persistence was observed. LSC quantitation provides a novel biomarker of disease response and relapse in patients with AML treated with epigenetic therapies. New drugs that target this cellular population in vivo are required.

Systemic arterial compliance following ultra-marathon.

There is a growing interest in training for and competing in race distances that exceed the marathon; however, little is known regarding the vascular effects of participation in such prolonged events, which last multiple consecutive hours. There exists some evidence that cardiovascular function may be impaired following extreme prolonged exercise, but at present, only cardiac function has been specifically examined following exposure to this nature of exercise. The primary purpose of this study was to characterize the acute effects of participation in an ultra-marathon on resting systemic arterial compliance. Arterial compliance and various resting cardiovascular indices were collected at rest from 26 healthy ultra-marathon competitors using applanation tonometry (HDI CR-2000) before and after participation in a mountain trail running foot race ranging from 120-195 km which required between 20-40 continuous hours (31.2±6.8 h) to complete. There was no significant change in small artery compliance from baseline to post race follow-up (8.5±3.4-7.7±8.2 mL/mmHgx100, p=0.65), but large artery compliance decreased from 16.1±4.4 to 13.5±3.8 mL/mmHgx10 (p=0.003). Participation in extreme endurance exercise of prolonged duration was associated with acute reductions in large artery compliance, but the time course of this effect remains to be elucidated.

The detection of post-traumatic angle recession by gonioscopy in a population-based glaucoma survey.

BACKGROUND: Blunt trauma is responsible for most eye injuries in urban populations. Anterior chamber angle recession has been reported to be the most common sign of previous blunt trauma to the eye. The cumulative lifetime prevalence of post-traumatic angle recession has not been reported previously, and the relation between angle recession and glaucoma in a population-based setting is unknown. METHODS: As part of a population-based glaucoma survey, gonioscopy was performed on 987 (82.7%) of 1194 inhabitants of the village of Mamre, near Cape Town, South Africa, who were 40 years of age or older. RESULTS: Some degree of angle recession was identified in one eye of 60 people and in both eyes of 86 people. Men were affected more than three times as often as women in the fifth, sixth, and seventh decades. The cumulative lifetime prevalence of angle recession in this community was 14.6%. The prevalence of glaucoma in people with angle recession was 5.5% (8/146). Of 87 eyes with 360 degrees of angle recession, only 7 (8.0%) had glaucoma. Excessive alcohol consumption was significantly related to the presence of angle recession in women (P < 0.001). The prevalence of monocular blindness due to trauma was 2.5% (25/987). CONCLUSION: Although the importance of the study may be limited to this community, the findings suggest that future population-based studies of ocular trauma should include gonioscopy on all individuals examined. Secondary glaucomas, especially those related to trauma, should be screened for in developing countries when trying to establish the prevalence of potential visual loss from glaucoma.

Corticosteroids in corneal graft rejection: double versus single pulse therapy.

In a previous study, we reported that a single intravenous pulse of 500 mg of methylprednisolone is more effective than oral prednisone in reversing rejection in patients who present early with severe endothelial rejection. In addition, the grafts that survived were less likely to undergo a further rejection episode. This study investigates whether a second pulse given at 24 h (n = 31) or at 48 h (n = 29) has any advantage over a single pulse (n = 30). All three treatment regimens were equally effective in reversing rejection (74.2%, 79.3%, and 83.3%, respectively), and there was no statistical difference in graft survival. An additional pulse therefore appears to offer no advantage over single pulse therapy. In high-risk grafts, current therapy has been reported to be less effective in reversing graft rejection. This study showed no significant difference between high- and low-risk grafts, indicating that pulse therapy may have a particular role in these difficult cases.

The prevalence of primary angle closure glaucoma and open angle glaucoma in Mamre, western Cape, South Africa.

OBJECTIVE: To determine the prevalence of primary angle closure glaucoma in the so-called Cape people of mixed ethnic background. DESIGN: A population-based prevalence study. SETTING: Mamre, a village near Cape Town, South Africa. PARTICIPANTS: Individuals aged 40 years or older. Historically, their ancestors were Southeast Asians and indigenous Africans and, to a lesser extent, Europeans. Of a total of 1194 people, 987 (82.7%) were examined. MAIN OUTCOME MEASURE: Primary angle closure glaucoma was diagnosed in individuals with previous acute or intermittent symptoms of angle closure and in individuals with an "occludable" angle and an intraocular pressure of greater than 21 mm Hg or a glaucomatous visual field. MAIN RESULTS: An age-related trend toward hypermetropia was found, which was greatest in women older than age 50 years. Gonioscopy identified Shaffer grade 1 angles in 89 (9%) of 987 subjects. The prevalence of primary angle closure glaucoma was 2.3% (23 subjects) and increased with age in both sexes. Women were affected more than four times as often as men and the sex difference persisted across all age groups. In comparison, the prevalence of primary open angle glaucoma was 1.5% (15 subjects). Primary glaucoma (angle closure plus open angle) was the leading cause of bilateral blindness in the community, with a prevalence rate of 0.5% (five subjects). CONCLUSIONS: This study identified primary angle closure glaucoma as a significant public health problem in the Western Cape Province. Because of the ethnic back-ground of the people studied, these findings may also apply to the populations of Southeast Asia.

The effects of unconscious presentation of information on therapist conceptualizations, intentions, and responses.

This article presents a study of the effects of tachistoscopic presentation of affective words on subjects' conceptualizations, intentions, and responses to a simulated client. The participants, 36 counseling students, were assigned randomly to one of the following treatments: subliminal presentation of negative emotional concepts; subliminal presentation of positive emotional concepts; supraliminal presentation of positive emotional concepts. After the tachistoscopic presentations, all subjects were exposed to a simulated client, whom they were asked to evaluate, respond to, and report the cognitive intentions that guided their responses. Significant effects were found in the subliminal presentation of positive emotional concepts on subjects' conceptualizations, intentions, and responses. Some significant effects also were found for the supraliminal presentation, but only for the client evaluation measure.

Differentiation of alcoholic from brain-damaged patients by the Psychiatric-Organic (P-O) special scale of the MMPI.

The value of the Psychiatric-Organic (P-O) special scale of the MMPI for the separation of the alcoholic from brain-damaged patients was investigated. A sample of 55 males containing 30 brain-damaged patients and 25 alcoholic patients, was administered the MMPI. Statistical analysis yielded significant difference. Clinical implications of the results are discussed.