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Background: This follow-up study compared patients with schizophrenia with co-occurring substance use disorder to patients with schizophrenia and no substance use disorder.Aims: To investigate the prognostic significance of the effects of substance use disorders on the course of schizophrenia.Method: Patients with schizophrenia and co-occurring substance use disorder (n=107), and patients with schizophrenia only (n=119) were followed over a 15-year period through the use of national hospitalization registers, data for time and cause of death, and data for homelessness or institutionalization.Results: The median length of psychiatric hospitalization was 12 days for a patient with dual diagnosis, and 21 days for patients with schizophrenia only. Patients with dual diagnosis displayed a significantly elevated usage of all types of hospital contacts except inpatient treatment for non-psychiatric disorders. In three types of hospital contacts, patients with co-occurring substance use had approximately two to three times as many hospitalizations as did patients with schizophrenia only. Rates for homelessness and institutionalization were similar in both groups. Patients with a dual diagnosis were also significantly more at risk of dying during follow-up than were patients with schizophrenia only.Conclusions: The findings suggest that the long-term course of schizophrenia is considerably more severe in patients who have a dual diagnosis compared to patients with schizophrenia only Substance use disorders have a substantial impact on the hospitalization rates of patients with schizophrenia, as well as on life expectancy. Patients with co-morbid substance abuse are more likely to be admitted for treatment during a given year although they have briefer contact with treatment. [ABSTRACT FROM AUTHOR]

Background: Psoriatic arthritis can involve several domains. Due to its multifaceted nature and its frequent comorbidities such as depression, obesity, osteoarthritis and fibromyalgia, it is difficult to monitor these patients because the clinical scores involve subjective data. High-resolution ultrasound probes allowed the evaluation of more superficial structures, such as the nails and their synovio-entheseal framework, in close relationship with the enthesis of the distal extensor digitorum tendon. Nail ultrasound studies vary in terms of the parameters and fingers studied and in their findings., Objectives: To describe the most significant sonographic nail changes and the most affected fingers in psoriatic arthritis and to verify the association of nail ultrasound findings with clinical scores (nail psoriasis severity index (NAPSI), ankylosing spondylitis disease activity score with C-reactive protein (ASDAS-CRP), minimal disease activity (MDA), disease activity index for psoriatic arthritis (DAPSA))., Methods: This was a cross-sectional study with 52 patients with psoriatic arthritis at the Hospital de Clínicas do Paraná and 50 controls. A total of 1016 nails were analyzed (517 from patients with psoriatic arthritis and 499 from controls). Ultrasonography of the nails of the 10 fingers was performed to assess the trilaminar appearance, measure the distance from the nail bed, identify synovitis of the distal interphalangeal joints and the presence of a power Doppler signal from the nail matrix/nail bed. The captured images were independently evaluated by a rheumatologist with expertise in musculoskeletal ultrasound. Data analysis was performed using IBM SPSS Statistics v.28.0.0 software, and the association of nail plate changes, nail bed distance and power Doppler signal with the NAPSI, DAPSA, MDA and ASDAS-PCR were calculated. Spearman correlation coefficients were estimated to analyze the correlations between pairs of quantitative variables. Student's t test and the Mann‒Whitney U test were used to compare quantitative variables, and Fisher's exact test was used to compare categorical variables between patients and controls. The nonparametric Mann‒Whitney U and Kruskal‒Wallis tests were used to compare groups according to the MDA or DAPSA classification., Results: The Doppler signal of the nail matrix and nail bed was more frequently identified in patients (44.2%) than in controls (6%), and the difference in the mean power Doppler signal between the two groups was significant (p < 0.001). Changes in the nail plate were more common in the right thumb (44.2%), left thumb (36.5%) and second finger on the right hand (32.7%). The number of fingers with nail plate changes, enthesitis, paratendinitis, grayscale synovitis and DIP involvement in the distal interphalangeal joints was higher among patients with psoriatic arthritis (p < 0.001). There were found some correlations between US findings and clinical scores: ultrasound nail involvement and the NAPSI score (p = 0.034), the number of fingers and mean change in the nail plate and the ASDAS-CRP (p = 0.030). DAPSA (remission/low activity versus moderate/high activity) was associated to the mean change in the nail plate (p < 0.013)., Conclusions: Nail ultrasound has the potential to assist in the capturing of the actual disease activity status in patients with psoriatic arthritis., (© 2024. The Author(s).)

Transcutaneous auricular vagus nerve stimulation (taVNS) is a non-invasive neuromodulation technique that modulates the noradrenergic activity of the locus coeruleus (LC). Yet, there is still uncertainty about the most effective stimulation and reliable outcome parameters. In a double blind, sham-controlled study including a sample of healthy young individuals ( N = 29), we compared a shorter (3.4 s) and a longer (30 s) stimulation duration and investigated the effects of taVNS (real vs. sham) on saliva samples (alpha amylase and cortisol concentration), pupil (pupillary light reflex and pupil size at rest) and EEG data (alpha and theta activity at rest, ERPs for No-Go signals), and cognitive tasks (Go/No-Go and Stop Signal Tasks). Salivary alpha amylase concentration was significantly increased in the real as compared to sham stimulation for the 30 s stimulation condition. In the 3.4 s stimulation condition, we found prolonged reaction times and increased error rates in the Go/No-Go task and increased maximum acceleration in the pupillary light reflex. For the other outcomes, no significant differences were found. Our results show that prolonged stimulation increases salivary alpha-amylase, which was expected from the functional properties of the LC. The finding of longer response times to short taVNS stimulation was not expected and cannot be explained by an increase in LC activity. We also discuss the difficulties in assessing pupil size as an expression of taVNS-mediated LC functional changes.

The basal structure of the bacterial flagellum includes a membrane embedded MS-ring (formed by multiple copies of FliF) and a cytoplasmic C-ring (composed of proteins FliG, FliM and FliN). The SRP-type GTPase FlhF is required for directing the initial flagellar protein FliF to the cell pole, but the mechanisms are unclear. Here, we show that FlhF anchors developing flagellar structures to the polar landmark protein HubP/FimV, thereby restricting their formation to the cell pole. Specifically, the GTPase domain of FlhF interacts with HubP, while a structured domain at the N-terminus of FlhF binds to FliG. FlhF-bound FliG subsequently engages with the MS-ring protein FliF. Thus, the interaction of FlhF with HubP and FliG recruits a FliF-FliG complex to the cell pole. In addition, the modulation of FlhF activity by the MinD-type ATPase FlhG controls the interaction of FliG with FliM-FliN, thereby regulating the progression of flagellar assembly at the pole., (© 2024. The Author(s).)

Sea urchins are primary herbivores on coral reefs, regulating algal biomass and facilitating coral settlement and growth. 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 Recurring mass mortality events (MMEs) of Diadema species Gray, 1825 have been recorded globally, 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 the most notorious and ecologically significant of which occurred in the Caribbean in 1983, 14 , 17 , 19 , 20 contributing to the shift from coral to algal-dominated ecosystems. 17 , 24 , 25 Recently, first evidence of Diadema setosum mass mortality was reported from the eastern Mediterranean Sea. 23 Here, we report extensive mass mortalities of several diadematoid species inhabiting the Red Sea and Western Indian Ocean (WIO) 26 , 27 , 28 including first evidence of mortalities in the genus Echinothrix Peters, 1853. Mortalities initiated in the Gulf of Aqaba on December 2022 and span the Red Sea, the Gulf of Oman, and the Western Indian Ocean (Réunion Island), with population declines reaching 100% at some sites. Infected individuals are characterized by spine loss and tissue necrosis, resulting in exposed skeletons (i.e., tests) and mortality. Molecular diagnostics of the 18S rRNA gene confirm the presence of a waterborne scuticociliate protozoan most closely related to Philaster apodigitiformis in infected specimens-identical to the pathogen found in the 2022 Caribbean mass mortality of Diadema antillarum. 13 , 15 , 18 Collapse of these key benthic grazers in the Red Sea and Western Indian Ocean may lead to algal dominance over corals, threatening the stability of coral reefs on a regional scale. 29 , 30 , 31 , 32 We issue a warning regarding the further expansion of mortalities and call for immediate monitoring and conservation efforts for these key ecological species., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Excessive mind wandering (MW) contributes to the development and maintenance of psychiatric disorders. Previous studies have suggested that auditory beat stimulation may represent a method enabling a reduction of MW. However, little is known about how different auditory stimulation conditions are subjectively perceived and whether this perception is in turn related to changes in subjective states, behavioral measures of attention and MW. In the present study, we therefore investigated MW under auditory beat stimulation and control conditions using experience sampling during a sustained attention to response task (SART). The subjective perception of the stimulation conditions, as well as changes in anxiety, stress and negative mood after versus before stimulation were assessed via visual-analog scales. Results showed that any auditory stimulation applied during the SART was perceived as more distracting, disturbing, uncomfortable and tiring than silence and was related to more pronounced increases of stress and negative mood. Importantly, the perception of the auditory conditions as disturbing was directly correlated with MW propensity. Additionally, distracting, disturbing and uncomfortable perceptions predicted negative mood. In turn, negative mood was inversely correlated with response accuracy for target stimuli, a behavioral indicator of MW. In summary, our data show that MW and attentional performance are affected by the adverse perception of auditory stimulation, and that this influence may be mediated by changes in mood., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Schmidt, Chaieb, Derner, Reber and Fell.)

Little is known about the effects of woody plant encroachment-a recent but pervasive phenomenon-on the hydraulic properties of bedrock substrates. Recent work using stream solute concentrations paired with weathering models suggests that woody plant encroachment accelerates limestone weathering. In this field study, we evaluate this hypothesis by examining bedrock in the Edwards Plateau, an extensive karst landscape in Central Texas. We compared a site that has been heavily encroached by woody plants (mainly Quercus fusiformis and Juniperus ashei), with an adjacent site that has been maintained free of encroachment for the past eight decades. Both sites share the same bedrock, as confirmed by trenching, and originally had very few trees, which enabled us to evaluate how encroachment impacted the evolution of hydraulic properties over a period of no more than 80 years. Using in situ permeability tests in boreholes drilled into the weathered bedrock, we found that the mean saturated hydraulic conductivity of the bedrock was higher-by an order of magnitude-beneath woody plants than in the areas where woody plants have been continuously suppressed. Additionally, woody plant encroachment was associated with greater regolith thickness, greater plant rooting depths, significantly lower rock hardness, and a 24-44% increase in limestone matrix porosity. These findings are strong indicators that woody plant encroachment enhances bedrock weathering, thereby amplifying its permeability-a cycle of mutual reinforcement with the potential for substantial changes within a few decades. Given the importance of shallow bedrock for ecohydrological and biogeochemical processes, the broader impacts of woody plant encroachment on weathering rates and permeability warrant further investigation., (© 2023. Springer Nature Limited.)

Background: Many patients with temporomandibular disorders (TMD) find it difficult to undergo dental care due to challenges caused by their condition, previous temporomandibular joint surgery or invasive dental procedures, and the impact of comorbid conditions. Managing routine dental care for some patients with TMD can be seen as challenging by some dental practitioners., Objective: The objective of this study was to work with patients experiencing TMD and clinicians to co-produce recommendations aimed at helping general dentists to provide routine dental care for patients with TMD., Methods: A modified Delphi process was used to co-produce recommendations. Six patients experiencing TMD, patient advocates and seven clinicians took part, including international TMD clinicians. Two meetings were held with patient participants, mediated by a trained facilitator. Recommendations suggested by patient participants were distributed to clinicians who were asked to add additional suggestions, but not to modify patients' recommendations unless to aid clarity. Additional themes were identified from the existing literature, and the recommendations were then reviewed by the International Network for Orofacial Pain and Related Disorders Methodology (INfORM) consortium., Results: Recommendations were given to support patients before, during and after dental treatment. Participants identified specific and practical recommendations to help patients with TMD receive routine dental care, but also emphasised the need for professionals to listen sensitively to patients' concerns and work with patients in an empathetic and non-judgmental way., Conclusion: These recommendations, co-developed with patients experiencing TMD, should help dental professionals to provide supportive general dental care for patients with TMD., (© 2023 The Authors. Journal of Oral Rehabilitation published by John Wiley & Sons Ltd.)

The sea urchin Diadema setosum is an ecological key species across its range, particularly on coral reefs. In 2006 D. setosum was first observed in the Mediterranean Sea, and since, it has proliferated to occupy the entire Levantine Basin. Here we report the mass mortality of the invasive D. setosum in the Mediterranean Sea. This is the first report of D. setosum mass mortality. The mortality spans over 1000 km along the Levantine coast of Greece and Turkey. The current mortality shows similar pathologies to previously reported Diadema mass mortality events, suggesting pathogenic infection as the cause of mortalities. Maritime transport, local currents, and fish predation of infected individuals may distribute pathogens at varying geographical scales. Due to the proximity of the Levantine Basin to the Red Sea, the risk of pathogen transport to the native Red Sea D. setosum population is imminent-with potentially catastrophic consequences., Competing Interests: We declare we have no competing interests., (© 2023 The Authors.)

A revised version of the Multicultural Ideology Scale (rMCI) is currently being developed to measure endorsement of multiculturalism in different cultural contexts. This study, which is part of a wider cross-cultural research project, presents the first assessment of the rMCI scale in the German language. The measure aims to cover several attitudinal dimensions of multiculturalism, relevant to the integration of different ethnocultural groups: Cultural Maintenance, Equity/Inclusion, Social interaction, Essentialistic Boundaries, Extent of Differences, and Consequences of Diversity. Two independent datasets were acquired from Germany ( N  = 382) and Luxembourg ( N  = 148) to estimate the factor structure of the rMCI using different confirmatory factor analysis techniques. The findings suggest that a four-factor solution, including Cultural Maintenance, Equity/Inclusion, Social interaction, and Consequences of Diversity, was the best fit for the data. Most of these subscales demonstrated adequate psychometric properties (internal consistency, convergent, and discriminant validity). The four-factor model of the rMCI was partially invariant across the two ethnic groups and full measurement invariance was established across gender.

3D printing has been explored as an emerging technology for the development of versatile and printable materials for drug delivery. However, the alliance of 3D printing and nanomaterials has, to date, been little explored in pharmaceutics. Herein, a mesoporous silica with nanostructured pores, SBA-15, was used as a drug carrier for triamcinolone acetonide, a hydrophobic drug, with the aim of incorporating the drug formulation in a hydrophilic printable ink. The adsorption of the drug in the SBA-15 pores was confirmed by the decrease in its surface area and pore volume, along with an increase in the apparent aqueous solubility of triamcinolone acetonide, as shown by in vitro release studies. Thereafter, a hydrophilic ink composed of carboxymethyl cellulose containing drug-loaded SBA-15 was formulated and 3D printed as hydrophilic polymeric film using the semisolid extrusion technique (SSE). The 3D printed films showed complete drug release after 12 h, and the presence of the triamcinolone acetonide-loaded SBA-15 improved their in vitro mucoadhesion, suggesting their promising application in oral mucosa treatments. Besides representing an innovative platform to develop water-based mucoadhesive formulations containing a hydrophobic drug, this is the first report proposing the development of SSE 3D printed nanomedicines containing drug-loaded mesoporous silica., (Copyright © 2022 Elsevier B.V. All rights reserved.)

In contrast to most industrialized countries, maternal mortality in the USA is rising. Cardiovascular disease, both acquired heart disease (e.g. coronary disease, arrhythmias, and heart failure), as well as congenital heart disease survivors, are all potentially important factors in explaining this worrisome trend. Increase in acquired cardiac disease is likely attributable to greater rates of obesity, diabetes, hypertension, and an increase in the incidence of advanced maternal age, while congenital heart disease in pregnancy is increasing due to advances in pediatric cardiovascular surgery. Despite the growing cardiovascular risk of pregnant women, most obstetricians and cardiologists have limited experience in caring for women with heart disease. Accordingly, management is largely guided by expert opinion likely to vary greatly across centers. To address these challenges, a multidisciplinary approach to care that includes both cardiologists and obstetricians could leverage the knowledge of both specialties and support streamlined communication between the patient and her providers. Our experience highlights the necessary components and essential infrastructure for building a center of excellence in treating pregnant women with heart disease. Condensation: A guide for creating a center of excellence for prenatal care for women with cardiovascular disease. The problem: Cardiac disease is the leading cause of maternal mortality, and pregnancies affected by cardiac disease continue to rise, both congenital and acquired. The solution: Maternal fetal medicine, obstetricians, and cardiologists can join together in tertiary facilities to create Maternal Cardiac Centers of Excellence to provide multidisciplinary, structured care for these high-risk patients.

In the past several decades, field studies have shown that woody plants can access substantial volumes of water from the pores and fractures of bedrock 1-3 . If, like soil moisture, bedrock water storage serves as an important source of plant-available water, then conceptual paradigms regarding water and carbon cycling may need to be revised to incorporate bedrock properties and processes 4-6 . Here we present a lower-bound estimate of the contribution of bedrock water storage to transpiration across the continental United States using distributed, publicly available datasets. Temporal and spatial patterns of bedrock water use across the continental United States indicate that woody plants extensively access bedrock water for transpiration. Plants across diverse climates and biomes access bedrock water routinely and not just during extreme drought conditions. On an annual basis in California, the volumes of bedrock water transpiration exceed the volumes of water stored in human-made reservoirs, and woody vegetation that accesses bedrock water accounts for over 50% of the aboveground carbon stocks in the state. Our findings indicate that plants commonly access rock moisture, as opposed to groundwater, from bedrock and that, like soil moisture, rock moisture is a critical component of terrestrial water and carbon cycling., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)

Inducible expression of neoantigens in mice would enable the study of endogenous antigen-specific naïve T cell responses in disease and infection, but has been difficult to generate because leaky antigen expression in the thymus results in central T cell tolerance. Here we develop inversion-induced joined neoantigen (NINJA), using RNA splicing, DNA recombination and three levels of regulation to prevent leakiness and allow tight control over neoantigen expression. We apply NINJA to create tumor cell lines with inducible neoantigen expression, which could be used to study antitumor immunity. We also show that the genetic regulation in NINJA mice bypasses central and peripheral tolerance mechanisms and allows for robust endogenous CD8 and CD4 T cell responses on neoantigen induction in peripheral tissues. NINJA will enable studies of how T cells respond to defined neoantigens in the context of peripheral tolerance, transplantation, autoimmune diseases and cancer.

The economic viability of producing lignin by thermal-enzymatic treatment in a high-pressure fixed-bed reactor is investigated for the first time. In this direction, different advantages (e.g. recovery of low-odor sulfur-free lignin, high process flexibility) and disadvantages (e.g. high investment for high-pressure equipment) of this technology are considered. Regarding process flexibility, four different operating modes (i.e. flow through, circulation) are investigated by varying process parameters and applications of the C5 sugar fraction. Therefore, a combined modelling approach is applied by using overall biorefinery models and a predictive fixed-bed model. At optimum process conditions, lignin can be produced at a competitive price (395 €/t Lignin ) when comparing to other technical lignins. This result is achieved by using the fixed-bed reactor only for thermal treatment, with the water consumption being the most important factor affecting the cost of lignin production. Compared to that the C5 sugar recovery of the pretreatment is negligible., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

In order to minimize the environmental impacts of growing population, progressive exploitation of fossil resources and negative consequences of climate change the politically intended goal is to successively transform our primarily oil-based into a bio-based economy. Hence, one goal is to significantly reduce the utilization of fossil resources by increasing the use of renewable energy and resources (i.e. biomass) and the efficiency of their conversion processes. Including existing technologies into the development of future concepts could accelerate the transition to a bio-economy. As one solution integrated biorefinery concepts based on agricultural biogas plants are discussed, which convert biomass with minimal energy consumption to a multitude of products without generating waste. However, they still have huge potential in terms of increased biomass utilization. In that context, catch crops offer interesting opportunities as a substrate for those biorefineries, since they support soil regeneration while generating additional products for the bio-economy without increasing land use. In this study a selection of significant indicators was chosen in order to determine the environmental effectivity and economic efficiency of these biorefinery concepts by a systematic assessment of possible process schemes. Thus within this study the usability of the chosen indicators and the potential of catch crops in advanced biorefineries is assessed., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objectives: To examine whether early secondary repair of labial tears, 1st and 2nd degree perineal lacerations and episiotomies provided an anatomically acceptable result., Study Design: A retrospective analysis of 126 women undergoing an early secondary repair of birth lacerations not involving the sphincter complex within 21 days postpartum. Patients were included from 1 January 2014 to 11 August 2017 at Aarhus University Hospital, Denmark. Photo documentation of the wound was available just before the early secondary repair and at the follow-up visit after the repair. Photos were evaluated by two trained Urogynaecological Consultants., Main Outcome Measures: Whether the anatomic result of the early secondary repair was acceptable based on photo documentation., Results: Early secondary repair was performed by a specialised team of midwives in 94.4% and by doctors in 5.6% of the cases. In all, 72.2% were 2nd degree perineal lacerations. The most common indications for early secondary repair were wound dehiscence (55.3%) and suboptimal primary repair (34.1%). At the follow-up clinical examination seven days after the early secondary repair, the result was considered anatomically acceptable in 67.5% of the cases, not optimal in 22.2% and not possible to evaluate in 10.3% of the cases. In 7.9% of the cases, wound infection was suspected after the early secondary repair., Conclusions: Based on photo documentation, early secondary repair of birth lacerations not involving the sphincter complex provides an anatomically acceptable result in the majority of cases without risk of serious complications., (Copyright © 2018. Published by Elsevier India Pvt Ltd.)

Objective: Psychotherapy training is mandatory for physicians to qualify as psychiatrists in Denmark. Evidence for the effectiveness of psychotherapy has increased, and psychotherapy is increasingly included in international treatment guidelines. The authors investigated how psychiatrists in training in Denmark evaluate the opportunities to practice psychotherapy in their training and the quality of the supervision they receive in psychotherapy training, particularly for cognitive behavioral therapy (CBT)., Method: The authors conducted a survey regarding psychotherapy training and CBT supervision among psychiatrists in training at Danish psychiatric specialist training courses. They investigated respondents' interest and experience in psychotherapy and respondents' views on the relevance and feasibility of performing psychotherapy and receiving supervision in their psychiatry training., Results: Eighty-eight percent of the psychiatrists in training found psychotherapy to be a relevant part of their training; however, 77 % found it difficult to find time to practice psychotherapy and 44 % felt that practicing psychotherapy was a strain on their employer. Thirty-six percent and 53 %, respectively, had difficulties securing psychodynamic and CBT supervision. In CBT supervision, more than 60 % reported supervision that appeared to be below the expected CBT supervision standard and often so much below it might not qualify as CBT supervision., Conclusions: There is a need to focus on how to better integrate psychotherapy and supervision in the Danish psychiatric training program. Good CBT supervision may be lacking, and a way to ensure high-quality supervision is required.

Aims: To generate single spore lines of a population of bacterial parasite of root-knot nematode (RKN), Pasteuria penetrans, isolated from Florida and examine genotypic variation and virulence characteristics exist within the population., Methods and Results: Six single spore lines (SSP), 16SSP, 17SSP, 18SSP, 25SSP, 26SSP and 30SSP were generated. Genetic variability was evaluated by comparing single-nucleotide polymorphisms (SNPs) in six protein-coding genes and the 16S rRNA gene. An average of one SNP was observed for every 69 bp in the 16S rRNA, whereas no SNPs were observed in the protein-coding sequences. Hierarchical cluster analysis of 16S rRNA sequences placed the clones into three distinct clades. Bio-efficacy analysis revealed significant heterogeneity in the level virulence and host specificity between the individual clones., Conclusions: The SNP markers developed to the 5' hypervariable region of the 16S rRNA gene may be useful in biotype differentiation within a population of P. penetrans., Significance and Impact of the Study: This study demonstrates an efficient method for generating single spore lines of P. penetrans and gives a deep insight into genetic heterogeneity and varying level of virulence exists within a population parasitizing a specific Meloidogyne sp. host. The results also suggest that the application of generalist spore lines in nematode management may achieve broad RKN control., (© 2016 The Society for Applied Microbiology.)

Previous research has shown that multi-unit housing (MUH) residents are at risk of secondhand smoke (SHS) exposure, which can transfer between units. The purpose of this study was to determine SHS exposure and examine attitudes towards smoking policies among public housing authority (PHA) residents in rural and tribal settings. A self-administered questionnaire was completed by 895 adult tenants (41 % response rate) living in PHA multiunit buildings in Montana in 2013. Our primary outcome was tenant support of smoke-free policies; our secondary outcome was exacerbation of child asthma symptoms due to SHS exposure. In 2014, we used multiple logistic regression models to test associations between independent variables and outcomes of interest. The majority (80.6 %) of respondents supported having a smoke-free policy in their building, with support being significantly higher among nonsmokers [adjusted odds ratio (aOR) 4.2, 95 % confidence interval (CI) 1.5-11.6] and among residents living with children (aOR 2.9, 95 % CI 1.3-6.2). Tribal residents were as likely to support smoke-free policies as non-tribal residents (aOR 1.4; 95 % CI 0.5-4.0). Over half (56.5 %) of respondents reported SHS exposure in their home; residents in a building with no smoke-free policy in place were significantly more likely to report exposure (aOR 3.5, 95 % CI 2.2-5.5). SHS exposure was not significantly associated with asthma symptoms. There is a significant reduction in exposure to SHS in facilities with smoke-free policies and there is strong support for such policies by both tribal and non-tribal MUH residents. Opportunities exist for smoke-free policy initiatives in rural and tribal settings.

Background: In today's consumer perception of industrial processes and food production, aspects like food quality, human health, environmental safety, and energy security have become the keywords. Therefore, much effort has been extended toward adding value to biowastes of agri-food industries through biorefinery processing approaches. This study focused, for the first time, on the valorization of tomato by-products of a Tunisian industry for the recovery of value-added compounds using biorefinery cascade processing., Results: The process integrated supercritical CO 2 extraction of carotenoids within the oil fractions from tomato seeds (TS) and tomato peels (TP), followed by a batch isolation of protein from the residues. The remaining lignocellulosic matter from both fractions was then submitted to a liquid hot water (LHW) hydrolysis. Supercritical CO 2 experiments extracted 5.79% oleoresin, 410.53 mg lycopene/kg, and 31.38 mg β-carotene/kg from TP and 26.29% oil, 27.84 mg lycopene/kg, and 5.25 mg β-carotene/kg from TS, on dry weights. Protein extraction yields, nearing 30% of the initial protein contents equal to 13.28% in TP and 39.26% in TS, revealed that TP and TS are a rich source of essential amino acids. LHW treatment run at 120-200 °C, 50 bar for 30 min showed that a temperature of 160 °C was the most convenient for cellulose and hemicellulose hydrolysis from TP and TS, while keeping the degradation products low., Conclusions: Results indicated that tomato by-products are not only a green source of lycopene-rich oleoresin and tomato seed oil (TSO) and of protein with good nutritional quality but also a source of lignocellulosic matter with potential for bioethanol production. This study would provide an important reference for the concept and the feasibility of the cascade fractionation of valuable compounds from tomato industrial by-products.Graphical abstractSchema of biorefinery cascade processing of tomato industrial by-products toward isolation of valuable fractions.

Airway epithelial cells (AEC) are the first line of defense against airborne infectious microbes and play an important role in regulating the local immune response. However, the interplay of epithelial cells and professional immune cells during both homeostasis and infection has only been partially studied. The present study was performed to determine how bronchial epithelial cells affect the activation of monocytes. Under healthy conditions, AECs were shown to inhibit reactivity of monocytes. We hypothesized that upon infection, monocytes might be released from inhibition by AECs. We report that direct contact of monocytes with unstimulated BEAS2B epithelial cells results in inhibition of TNF secretion by activated monocytes. In addition to the known soluble modulators, we show that cell contacts between epithelial cells and monocytes or macrophages also contribute to homeostatic inhibitory actions. We find AECs to express the inhibitory molecule PD-L1 and blockade of PD-L1 results in increased secretion of pro-inflammatory cytokines from monocytes. Contrary to the inhibitory activities during homeostasis, epithelial cells infected with Respiratory Syncitial Virus (RSV) induce a significant release of inhibition. However, release of inhibition was not due to modulation of PD-L1 expression in AECs. We conclude that airway epithelial cells control the reactivity of monocytes through direct and indirect interactions; however tonic inhibition can be reverted upon stimulation of AECs with RSV and thereof derived molecular patterns. The study confirms the important role of airway epithelial cells for local immune reactions., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Glycemic control remains suboptimal in youth with type 1 diabetes. Retrospective continuous glucose monitoring (CGM) has demonstrated utility in fine-tuning diabetes management by detecting postprandial hyperglycemia and hypoglycemia. In this study, we explored the process of 3-day masked CGM use, subsequent treatment recommendations, and impact on A1c in a clinic-based sample of youth with type 1 diabetes. Over 2 years, 122 youth were referred for masked CGM. Patients/families completed a diary of blood glucose levels, insulin doses, food intake, and exercise during CGM use. A1c was assessed pre- and 2-3 months post-CGM. Treatment recommendations were formulated using data from CGM reports and diaries. Mean age was 14.3 ± 3.9 years, diabetes duration was 7.5 ± 4.7 years, and A1c was 8.5 ± 1.1% (69 ± 12 mmol/mol); 61% were pump-treated. Patients received an average of 3.1 ± 1.1 treatment recommendations following review of the CGM report. Most (80%) received reinforcement of the importance of preprandial bolusing; 37% received a recommendation regarding advanced insulin management (use of combination boluses/attend to active insulin). Receipt of the latter recommendation was related to A1c improvement ≥0.5% (OR: 4.0, P < .001). Masked CGM offers opportunities to guide advanced insulin management (by injection or pump), which may yield A1c improvements in youth with type 1 diabetes., (© 2014 Diabetes Technology Society.)

Cancer immunoediting is a process by which immune cells, particularly lymphocytes of the adaptive immune system, protect the host from the development of cancer and alter tumour progression by driving the outgrowth of tumour cells with decreased sensitivity to immune attack. Carcinogen-induced mouse models of cancer have shown that primary tumour susceptibility is thereby enhanced in immune-compromised mice, whereas the capacity for such tumours to grow after transplantation into wild-type mice is reduced. However, many questions about the process of cancer immunoediting remain unanswered, in part because of the known antigenic complexity and heterogeneity of carcinogen-induced tumours. Here we adapted a genetically engineered, autochthonous mouse model of sarcomagenesis to investigate the process of cancer immunoediting. This system allows us to monitor the onset and growth of immunogenic and non-immunogenic tumours induced in situ that harbour identical genetic and histopathological characteristics. By comparing the development of such tumours in immune-competent mice with their development in mice with broad immunodeficiency or specific antigenic tolerance, we show that recognition of tumour-specific antigens by lymphocytes is critical for immunoediting against sarcomas. Furthermore, primary sarcomas were edited to become less immunogenic through the selective outgrowth of cells that were able to escape T lymphocyte attack. Loss of tumour antigen expression or presentation on major histocompatibility complex I was necessary and sufficient for this immunoediting process to occur. These results highlight the importance of tumour-specific-antigen expression in immune surveillance, and potentially, immunotherapy.

Drugs inhibiting the influenza A virus (IAV) neuraminidase (NA) are the cornerstone of anti-IAV chemotherapy and prophylaxis in man. Drug-resistant mutations in NA arise frequently in human isolates, limiting the therapeutic application of NA inhibitors. Here, we show that antibody-driven antigenic variation in one domain of the H1 hemagglutinin Sa site leads to compensatory mutations in NA, resulting in NA antigenic variation and acquisition of drug resistance. These findings indicate that influenza A virus resistance to NA inhibitors can potentially arise from antibody driven HA escape, confounding analysis of influenza NA evolution in nature.

Airway epithelial cells regulate immune reactivity of local dendritic cells (DCs), thus contributing to microenvironment homeostasis. In this study, we set out to identify factors that mediate this regulatory interaction. We show that tracheal epithelial cells secrete soluble factors that downregulate TNF-α and IL-12p40 secretion by bone marrow-derived DCs but upregulate IL-10 and arginase-1. Size exclusion chromatography identified small secreted molecules having high modulatory activity on DCs. We observed that airway tracheal epithelial cells constitutively release the lipid mediator PGE(2). Blocking the synthesis of PGs within airway epithelial cells relieved DCs from inhibition. Cyclooxygenase-2 was found to be expressed in primary tracheal epithelial cell cultures in vitro and in vivo as shown by microdissection of epithelial cells followed by real-time PCR. Paralleling these findings we observed that DCs treated with an antagonist for E-prostanoid 4 receptor as well as DCs lacking E-prostanoid 4 receptor showed reduced inhibition by airway epithelial cells with respect to secretion of proinflammatory cytokines measured by ELISA. Furthermore, PGE(2) mimicked the effects of epithelial cells on DCs. The results indicate that airway epithelial cell-derived PGE(2) contributes to the modulation of DCs under homeostatic conditions.

Neoantigens derived from somatic mutations in tumors may provide a critical link between the adaptive immune system and cancer. Here, we describe a system to introduce exogenous antigens into genetically engineered mouse lung cancers to mimic tumor neoantigens. We show that endogenous T cells respond to and infiltrate tumors, significantly delaying malignant progression. Despite continued antigen expression, T cell infiltration does not persist and tumors ultimately escape immune attack. Transplantation of cell lines derived from these lung tumors or prophylactic vaccination against the autochthonous tumors, however, results in rapid tumor eradication or selection of tumors that lose antigen expression. These results provide insight into the dynamic nature of the immune response to naturally arising tumors., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Rotylenchulus reniformis is one of 10 described species of reniform nematodes and is considered the most economically significant pest within the genus, parasitizing a variety of important agricultural crops. Rotylenchulus reniformis collected from cotton fields in the Southeastern US were observed to have the nematode parasitic bacterium Pasteuria attached to their cuticles. Challenge with a Pasteuria-specific monoclonal antibody in live immuno-fluorescent assay (IFA) confirmed the discovery of Pasteuria infecting R. reniformis. Scanning and transmission electron microscopy were employed to observe endospore ultrastructure and sporogenesis within the host. Pasteuria were observed to infect and complete their life-cycle in juvenile, male and female R. reniformis. Molecular analysis using Pasteuria species-specific and degenerate primers for 16s rRNA and spoII, and subsequent phylogenetic assessment, placed the Pasteuria associated with R. reniformis in a distinct clade within established assemblages for the Pasteuria infecting phytopathogenic nematodes. A global phylogenetic assessment of Pasteuria 16s rDNA using the Neighbor-Joining method resulted in a clear branch with 100% boot-strap support that effectively partitioned the Pasteuria infecting phytopathogenic nematodes from the Pasteuria associated with bacterivorous nematodes. Phylogenetic analysis of the R. reniformis Pasteuria and Pasteuria spp. parasitizing a number of economically important plant parasitic nematodes revealed that Pasteuria with different host specificities are closely related and likely constitute biotypes of the same species. This suggests host preference, and thus effective differentiation and classification are most likely predicated by an influential virulence determinant(s) that has yet to be elucidated. Pasteuria Pr3 endospores produced by in vitro fermentation demonstrated efficacy as a commercial bionematicide to control R. reniformis on cotton in pot tests, when applied as a seed treatment and in a granular formulation. Population control was comparable to a seed-applied nematicide/insecticide (thiodicarb/imidacloprid) at a seed coating application rate of 1.0 x 10(8) spores/seed.

Rapid antigenic evolution in the influenza A virus hemagglutinin precludes effective vaccination with existing vaccines. To understand this phenomenon, we passaged virus in mice immunized with influenza vaccine. Neutralizing antibodies selected mutants with single-amino acid hemagglutinin substitutions that increased virus binding to cell surface glycan receptors. Passaging these high-avidity binding mutants in naïve mice, but not immune mice, selected for additional hemagglutinin substitutions that decreased cellular receptor binding avidity. Analyzing a panel of monoclonal antibody hemagglutinin escape mutants revealed a positive correlation between receptor binding avidity and escape from polyclonal antibodies. We propose that in response to variation in neutralizing antibody pressure between individuals, influenza A virus evolves by adjusting receptor binding avidity via amino acid substitutions throughout the hemagglutinin globular domain, many of which simultaneously alter antigenicity.

The upper airways are prone to contact with pathogenic as well as non-pathogenic microbes, therefore immune recognition principles have to be tightly controlled. Here we show that human BEAS-2B bronchial epithelial cells inhibited secretion of the pro-inflammatory cytokines TNF-alpha and IL-12 by monocytes, macrophages and dendritic cells. This inhibitory effect could be transferred by supernatant of resting BEAS-2B cells and was also observed when primary murine tracheal epithelial cells were prepared. In contrast to inhibition of pro-inflammatory cytokine secretion epithelial cell-conditioned dendritic cells showed increased expression of IL-10 and arginase-1, thus displaying properties of alternative activation. Accordingly, Toll-like receptor-mediated up-regulation of CD40, CD86 and PD-L2 (CD273) on murine dendritic cells was reduced in the presence of bronchial epithelial cell supernatant. However, expression of negative regulatory PD-L1 (CD274) was increased and dendritic cell induced proliferation of T lymphocytes was diminished. Epithelial cells also showed a direct inhibitory effect on T lymphocyte proliferation and this was due to the constitutive secretion of TGF-beta by bronchial epithelial cells. Moreover, epithelial cell-conditioned T lymphocytes showed increased differentiation towards IL-10-producing Tr1 cells. The results indicate that bronchial epithelial cells induce a non-inflammatory microenvironment that regulates local immune homeostasis.

Objectives: To assess Danish surgical departments' attitudes to and clinical guidelines on the use of perioperative thrombo-prophylaxis (TP)., Materials and Methods: Questionnaires regarding TP were sent to Danish departments which performed major surgery in 2005. The questionnaires contained questions regarding the use of TP routinely after clinical guidelines for both elective and acute surgery, indications for the use of TP, the type of TP and the duration of the TP treatment. The results are registered descriptively. The results are compared with five previously performed surveys from 1981-2000., Results: In the five surveys from 1981 to 2000, 85%, 90%, 92%, 95%, 93% answered the questionnaire, in the present survey 88% answered. Since the beginning of these surveys, there has been a significant increase in the use of TP following clinical guidelines from approximately 50% in 1981 to 96% of the departments performing major surgery in 2005. In acute surgery we found a significant increase in the number of departments using TP following clinical guidelines from 43% in 2000 to 81% in 2005 (p<0.01 Fishers exact test). All departments that used TP employed low molecular weight heparin. The use of prolonged TP was sporadic., Conclusion: 30 years after the first evidence in favour of TP was published, 96% of surgical departments which perform major surgery use TP routinely based on international clinical recommendations. A large number of departments use TP in connection with acute surgery in comparison with previous surveys. A future task is more focus on the use of prolonged TP after major surgical procedures.

Pasteuria penetrans, an obligate endospore-forming parasite of Meloidogyne spp. (root knot nematodes), has been identified as a promising agent for biocontrol of these destructive agricultural crop pests. Pasteuria ramosa, an obligate parasite of water fleas (Daphnia spp.), has been shown to modulate cladoceran populations in natural ecosystems. Selected sporulation genes and an epitope associated with the spore envelope of these related species were compared. The sigE and spoIIAA/spoIIAB genes differentiate the two species to a greater extent than 16S rRNA and may serve as probes to differentiate the species. Single-nucleotide variations were observed in several conserved genes of five distinct populations of P. ramosa, and while most of these variations are silent single-nucleotide polymorphisms, a few result in conservative amino acid substitutions. A monoclonal antibody directed against an adhesin epitope present on P. penetrans P20 endospores, previously determined to be specific for Pasteuria spp. associated with several phytopathogenic nematodes, also detects an epitope associated with P. ramosa endospores. Immunoblotting provided patterns that differentiate P. ramosa from other Pasteuria spp. This monoclonal antibody thus provides a probe with which to detect and discriminate endospores of different Pasteuria spp. The presence of a shared adhesin epitope in two species with such ecologically distant hosts suggests that there is an ancient and ecologically significant recognition process in these endospore-forming bacilli that contributes to the virulence of both species in their respective hosts.

Pasteuria species are endospore-forming obligate bacterial parasites of soil-inhabiting nematodes and water-inhabiting cladocerans, e.g. water fleas, and are closely related to Bacillus spp. by 16S rRNA gene sequence. As naturally occurring bacteria, biotypes of Pasteuria penetrans are attractive candidates for the biocontrol of various Meloidogyne spp. (root-knot nematodes). Failure to culture these bacteria outside their hosts has prevented isolation of genomic DNA in quantities sufficient for identification of genes associated with host recognition and virulence. We have applied multiple-strand displacement amplification (MDA) to generate DNA for comparative genomics of biotypes exhibiting different host preferences. Using the genome of Bacillus subtilis as a paradigm, MDA allowed quantitative detection and sequencing of 12 marker genes from 2000 cells. Meloidogyne spp. infected with P. penetrans P20 or B4 contained single nucleotide polymorphisms (SNPs) in the spoIIAB gene that did not change the amino acid sequence, or that substituted amino acids with similar chemical properties. Individual nematodes infected with P. penetrans P20 or B4 contained SNPs in the spoIIAB gene sequenced in MDA-generated products. Detection of SNPs in the spoIIAB gene in a nematode indicates infection by more than one genotype, supporting the need to sequence genomes of Pasteuria spp. derived from single spore isolates.

Background: A significant proportion of patients with liver metastases from colorectal cancer have unresectable disease and we have used cytoablative treatment such as cryotherapy in some of these patients. We retrospectively reviewed patients who underwent hepatic cryotherapy for colorectal metastases and studied the effect of the clinical risk score (CRS) reported by Fong et al, which can predict survival following liver resection., Methods: A retrospective study was performed on patients who underwent hepatic cryotherapy between 1990 and 2000 in St George Hospital. There were 61 patients in this study and they were stratified into prognostic groups based on five preoperative CRS parameters: primary node positive, disease-free interval from primary to metastases < 12 months, number of hepatic tumours > 1, largest hepatic tumour > 5 cm and carcinoembryonic antigen level > 200 ng/mL. The median follow-up was 25 months., Results: The median survival was 26 months and the 3-year survival rate was 37%. Median survivals for patients with CRS scores 1 (13%), 2 (25%), 3 (53%), 4 (6%) and 5 (4%) were 37, 25, 30, 21 and 15 months, respectively (R2 = 0.81)., Conclusion: The CRS score can be used to predict outcome of hepatic cryotherapy, but the difference in survival between CRS 2, 3 and 4 is modest.

Background: Citations in scientific articles may tend to favor the views presented. We studied whether there is such reference bias in narrative review articles that discuss interventions against house dust mites for people with asthma., Design: Systematic review of reviews identified in a Medline search that expressed an opinion about the clinical effects of physical or chemical intervention methods., Main Outcome Measure: Positive bias was judged to have occurred if the reference list contained a higher proportion of trial references with significant results than among all trials available to the authors (published 2 years or more prior to the review)., Results: Seventy reviews were included, of which 63 (90%) recommended physical interventions. Forty-six reviews had trial references, 4 of these only to chemical interventions. In the remaining 42 reviews, reference bias was detected (P=2 x 10-8). The most quoted trial had only 7 patients per group, its claimed significant result was probably erroneous, and it did not report a clinical outcome. Intervention recommendations were often based on nonrandomized evidence, and the most quoted nonrandomized controlled study had included only 10 patients per group but claimed very positive results., Conclusion: The narrative review articles were severely biased, and their positive intervention recommendations are at variance with the systematic Cochrane Review on this topic and a recent very large trial of physical intervention, both of which failed to find an effect.

We report on the development of a PCR-based assay to detect Pasteuria penetrans infection of Meloidogyne arenaria in planta using specific primers for recently sequenced sigE, spoIIAB and atpF genes of P. penetrans biotype P20. Amplification of these genes in crude DNA extracts of ground tomato root galls using real-time kinetic PCR distinguished infected from uninfected M. arenaria race 1 by analysis of consensus thresholds for single copy genes. Fluorescent in situ hybridization (FISH) using the sigE primer sequence as a probe shows hybridization to P. penetrans cells in various stages of vegetative (pre-endospore) development. Ratios of gene copies for sigE and 16S rDNA were obtained for P. penetrans and compared to Bacillus subtilis as a genomic paradigm of endospore-forming bacteria. Phylogenetic analysis of the sigE gene from Gram-positive, endospore-forming bacteria finds P. penetrans most closely related Paenbacillus polymyxa. The sporulation genes (spo genes), particularly sigE, have sequence diversity that recommends them for species and biotype differentiation of the numerous Pasteuria isolates that infect a large number of plant-parasitic nematodes.

Purpose: Bowel preparations for colonoscopy have to balance the demand for adequate cleansing action of the bowel and patient acceptability. There has been no study comparing Picoprep-3 (sodium picosulfate), a relatively new product, to Fleet (sodium phosphate), a well-studied and widely used preparation. This study was designed to compare the efficacy and patient tolerance of these two bowel preparations for colonoscopy., Methods: A randomized, single-blinded, prospective trial was conducted. A total of 400 consecutive patients presenting for elective colonoscopy at St George Private Hospital during a 20-week period were randomly assigned to receive Picoprep-3 or Fleet. Patients were asked to record the effects of the preparation, noting tolerability, taste, and side effects. Two hundred patients were assigned to the Picoprep-3 group and 200 to the Fleet group. Surgeons were blinded to the preparation used and rated the quality of the bowel preparation on a scale of 1 to 5 (1 being the optimal score)., Results: Picoprep-3 was found to be better tolerated (P < 0.0001) and better tasting (P < 0.0001) than Fleet. Patients in the Picoprep-3 group reported significantly less nausea (P < 0.001), vomiting (P < 0.004), dizziness (P < 0.01), abdominal pains (P = 0.0005), and thirst (P < 0.0001) associated with the preparation. There was no significant difference in visualization of the colon between the two groups as judged by the two colonoscopists (P = 0.06)., Conclusions: Colonoscopy preparation with Picoprep-3 has similar efficacy but superior taste and tolerability compared with Fleet. Picoprep-3 caused less adverse side effects in the study population.

Pasteuria spp. include endospore-forming bacterial pathogens of cladoceran crustaceans and plant-parasitic nematodes. Propagation of these nematode pathogens requires attachment of soilborne endospores to nematode hosts, infection, growth, sporulation, and release of endospores to repeat the cycle of infection and propagation. The ability of these bacteria to suppress the levels of plant-parasitic nematodes in the field has made them particularly promising candidates for biocontrol of nematode diseases of plants. Genes encoding 16S ribosomal RNA have been sequenced for the cladoceran (water flea) parasite and type species, Pasteuria ramosa, and for Pasteuria spp. isolated from root-knot (Meloidogyne arenaria race 1 and Meloidogyne sp.), soybean cyst (Heterodera glycines), and sting (Belonolaimus longicaudatus) nematodes. These have provided a phylogenetic basis for their designation to a distinct clade within the family Alicyclobacillaceae of the gram-positive endospore-forming bacteria. Two apparent biotypes of P. penetrans demonstrating a host preference for different Meloidogyne spp. showed identical 16S rDNA sequences, suggesting host-recognition evolves within a given species. The sequences of genes encoding sporulation transcription factors, sigE and sigF, from P. penetrans biotype P-20 show different phylogenetic relationships to other endospore-forming bacteria, supporting their application to further discriminate Pasteuria spp. and biotypes. Distribution of an adhesin-associated epitope on polypeptides from different Pasteuria isolates provides an immunochemical approach to differentiate species and biotypes with specific host preferences. Application of bioinformatics to genomic data, as well as further characterization of the biochemical basis for host recognition, will facilitate development of Pasteuria spp. as benign alternatives to chemical nematicides.

Abstract Pasteuria penetrans is an obligate parasite of root-knot nematodes (Meloidogyne spp.) that has attracted significant attention as a promising biocontrol agent. The inability to culture P. penetrans has invoked the need for a quantitative detection capability to facilitate biocontrol studies. A chemical extraction method using urea, dithiothreitol and CHES buffer (UDC) is shown to release soluble endospore envelope antigen from endospores present in complex matrices, generating an extract that can be used to determine the levels of spores when compared to a standard in an enzyme-linked immunosorbent assay (ELISA) using a specific monoclonal antibody, MAb 2A41D10. Extractions can be performed in less than 1 h. Linear regression analysis routinely produced line fits with r(2)>0.90. Antigen extraction efficiency was not influenced by soil type. Three ELISA formats were analyzed for quantitative detection of P. penetrans endospores. A tertiary ELISA immunodetection system provided the lowest level of detection at approximately 300 spores per gram of soil. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis Western blots of soil extracts containing P. penetrans endospore antigen produced signature peptides bearing a common epitope characteristic of endospores of Pasteuria spp. MAb 2A41D10 was specific for Pasteuria spp. and did not react with extracts of Pasteuria-free soil or with spore extracts of native Gram-positive endospore-forming bacteria. Immunofluorescent microscopy revealed that MAb 2A41D10 recognizes an epitope uniformly distributed on the endospore surface. The development of a rapid extraction method and analysis of solubilized antigen by immunodetection has the potential for broad application in food and environmental microbiology.