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Background: As the global aging population expands, understanding older adults' preferences for place of death becomes pivotal in ensuring person-centered end-of-life care., Objective: This study aimed to investigate the influence of sociodemographic, health, and lifestyle-related factors on end-of-life care preferences of older adults in South Tyrol, Italy., Methods: Employing a cross-sectional design, a population-based survey was conducted with a stratified probabilistic sample of adults aged ≥ 75 years in South Tyrol (Autonomous Province of Bolzano/Bozen, Italy). From a randomly selected sample of 3,600 older adults, participants were invited to respond to a questionnaire that included items on older adults' preferences for place of death and socio-demographic and health- and lifestyle-related factors, including frailty (e.g., PRISMA-7). Descriptive and multinomial logistic regression analyses were performed., Results: The majority (55.3%) of the 1,695 older adults (participation rate: 47%) expressed a preference for dying at home and 12.7% indicated a desire for specialized end-of-life care in a healthcare facility. However, 27.9% refrained from disclosing their end-of-life care preferences. The factors influencing these preferences concerning the place of death included age, native language, educational level, living situation, and community. Compared to the preference of dying at own home or home of family or friends, older adults aged ≥ 85 years (OR = 0.57, P = 0.002) and living in an urban area (OR = 0.40, P < 0.001) were less likely to prefer dying at a hospital, palliative care unit, or hospice. Older adults living alone (OR = 1.90, P < 0.001), Italian-speaking (OR = 1.46, P = 0.03), and those with an educational level above high school (OR = 1.69, P = 0.002) were more likely to prefer dying at a hospital, palliative care unit, or hospice., Conclusions: End-of-life care preferences among older adults in South Tyrol were associated with socio-demographic, yet not health- and lifestyle-related factors. Recognizing and integrating these preferences is essential for developing, implementing, and evaluating interventions to promote advance care planning and provide effective, patient-centered end-of-life care., (© 2024. The Author(s).)

Celiac disease (CeD) is a chronic inflammatory disease of the small intestine, produced by ingesting dietary gluten products in susceptible people. Gluten causes an impairment of the mucosal surface and, consequently, an abnormal absorption of nutrients. Although malabsorption of essential nutrients is a major risk factor for various CeD-associated morbidities, genetic, immunological, and environmental factors also play an important role. The clinical presentation of CeD widely varies and can range from asymptomatic to full-blown symptoms due to the multi-system nature of CeD. The typical gastrointestinal (GI) manifestations of CeD include abdominal pain, diarrhea, bloating, and weight loss, but several hepatobiliary manifestations and a poor nutritional status have also been described. Currently, a gluten-free diet (GFD) is the only current evidence-based treatment that leads to the complete recovery of mucosal damage and the reversibility of its progression. Conversely, undiagnosed CeD might have severe consequences in children as well as in adult patients. This narrative overview aims to characterize the GI and hepatobiliary manifestations, nutritional deficiencies, and delayed pediatric development associated with unrecognized CeD in order to identify it promptly. Moreover, the role of GFD and how it could prevent long-term complications of CeD are described.

Celiac disease (CeD) diagnosis is a complicated process, requiring a multi-step procedure and a high level of clinical knowledge. Some scientific societies, mainly from Europe and North America, have proposed appropriate guidelines for the diagnosis and management of CeD. Since duodenal biopsy is particularly challenging for children, guidelines of the European Society for Pediatric Gastroenterology, Hepatology, and Nutrition, presented in 2012 and updated in 2020, have made it possible to avoid the biopsy in symptomatic pediatric patients with high levels of IgA anti-transglutaminase. Several parallel, similar studies in adults support the non-biopsy strategy. However, several pros and cons exist in applying such a strategy. The present narrative review reports the current evidence and the implication of omitting biopsy in the diagnosis of CeD in adults.

Millions of children and adults worldwide suffer from undiagnosed and untreated celiac disease (CeD). The clinical picture of CeD is highly heterogeneous and comprises manifestations that can affect almost the whole body. This narrative overview is aimed at characterizing diseases and complaints that are associated with unrecognized CeD and that frequently involve sites other than the gastrointestinal (G.I.) tract, i.e., dental, otorhinolaryngological, and ocular complications; skin and hair abnormalities; afflictions of the bones, joints, and muscles; cardiovascular affectations; kidney diseases; neuro-psychiatric disorders; and gynecological-obstetrical manifestations. The association between CeD and extra-GI manifestations is frequently overlooked, which leads to a delay in diagnosis. Most CeD-mediated disorders can be treated with a strict gluten-free diet (GFD), but some of them are irreversible unless CeD is diagnosed in time. Some manifestations can be classified as risk factors for CeD, and CeD screening tests for affected patients should be selectively considered. Apart from gastroenterologists, specialists in other medical disciplines can play an important role in identifying people with unrecognized CeD and may help prevent its progress and long-term complications. Further comprehensive investigations are necessary to clarify the pathogenesis of extra-GI manifestations and the effect of a GFD.

Background: Digital health technologies offer the potential to improve the daily lives of older adults, maintain their health efficiently, and allow aging in place. Despite increasing evidence of benefits and advantages, readiness for adopting digital interventions among older people remains underexplored., Objective: This study aims to explore the relationships between sociodemographic-, health-, and lifestyle-related factors and technology use in everyday life and community-dwelling older adults' readiness to adopt telemedicine, smartphones with texting apps, wearables, and robotics., Methods: This was a cross-sectional, population-based survey study with a stratified probabilistic sample of adults aged 75 years or older living in South Tyrol (autonomous province of Bolzano/Bozen, Italy). A random sample of 3600 community-dwelling older adults living at home was invited to complete a questionnaire including single items (older adults' readiness to use health technology) and scales (PRISMA-7; Program of Research on Integration of Services for the Maintenance of Autonomy). Descriptive and logistic regression analyses were performed to analyze the data., Results: In total, 1695 community-dwelling older adults completed the survey (for a response rate of 47%). In terms of potential digital health technology adoption, wearable devices were favored by 33.7% (n=571), telemedicine by 30.1% (n=510), smartphones and texting apps by 24.5% (n=416), and assistant robots by 13.7% (n=232). Sociodemographic-, health- and lifestyle-related factors, as well as the use of technology in everyday life, played a significant role in explaining readiness to adopt digital health technologies. For telemedicine, age ≥85 years (odds ratio [OR] 0.74, 95% CI 0.56-0.96), financial constraints (OR 0.68, 95% CI 0.49-0.95), and less than 2 hours of physical activity per week (OR 0.75, 95% CI 0.58-0.98) were associated with nonreadiness, while Italian-speaking participants (OR 1.54, 95% CI 1.16-2.05) and those regularly using computers (OR 1.74, 95% CI 1.16-2.60), smartphones (OR 1.69, 95% CI 1.22-2.35), and the internet (OR 2.26, 95% CI 1.47-3.49) reported readiness for adoption., Conclusions: Community-dwelling older adults display varied readiness toward the adoption of digital health technologies, influenced by age, mother tongue, living situation, financial resources, physical activity, and current use of technology. The findings underscore the need for tailored interventions and educational programs to boost digital health technology adoption among community-dwelling older adults., (©Dietmar Ausserhofer, Giuliano Piccoliori, Adolf Engl, Angelika Mahlknecht, Barbara Plagg, Verena Barbieri, Nicoletta Colletti, Stefano Lombardo, Timon Gärtner, Waltraud Tappeiner, Heike Wieser, Christian Josef Wiedermann. Originally published in JMIR Formative Research (https://formative.jmir.org), 30.04.2024.)

The diagnosis of celiac disease (CD) is complex and requires a multi-step procedure (symptoms, serology, duodenal biopsy, effect of a gluten-free diet, and optional genetic). The aim of the study was to contribute to the improvement of CD diagnosis by preparing a water-soluble gluten peptide fraction (called Solgluten) and by selecting gluten-specific enzyme-linked immunosorbent assays (ELISA) for the detection of gluten immunogenic gluten peptides (GIPs) in urine and blood serum spiked with Solgluten. Food-grade Solgluten was prepared by the extraction of a peptic digest of vital gluten with water, centrifugation, and freeze-drying. The process was relatively easy, repeatable, and cheap. The content of gliadin-derived GIPs was 491 mg/g. Solgluten was used as antigenic material to compare two competitive ELISA kits (R7021 and K3012) and two sandwich ELISA kits (M2114 and R7041) in their quality regarding the quantitation of GIPs in urine and blood serum. The quality parameters were the reactivity, sensitivity, coefficients of variation and determination, and curve shape. The evaluation of the kits showed a number of discrepancies in individual quality parameters measured in urine and serum. Due to the lowest limit of quantitation and the highest coefficient of determination, M2114 may be the first choice, while R7021 appeared to be less suitable because of the high coefficients of variation and unfavorable curve progression. The results set the stage for improving CD diagnosis by supplementing conventional blood tests with oral provocation with Solgluten and subsequent ELISA measurement of GIPs that could support the no-biopsy approach and by better assessing the effect of a gluten-free diet by monitoring adherence to the diet by measuring GIPs in urine and blood.

Celiac disease (CeD) is a chronic gluten-sensitive immune-mediated enteropathy characterized by numerous intestinal and extra-intestinal signs and symptoms. Among extra-intestinal manifestations, otorhinolaryngological (ORL) complaints in CeD are relatively rare and their relation to CeD is frequently overlooked by physicians. Recent studies underlined that the prevalence of recurrent aphthous stomatitis, aphthous ulcers, geographic tongue, and xerostomia was significantly increased in CeD patients compared with healthy individuals. However, data about the other oral manifestations of CeD, such as atrophic glossitis, glossodynia, angular cheilitis, and salivary abnormalities, are scanty. Further ORL conditions associated with CeD include sensorineural hearing loss, nasal abnormalities, and obstructive sleep apnea. Moreover, several esophageal disorders such as gastroesophageal reflux disease and eosinophilic esophagitis have been associated with CeD. The pathophysiological link between both ORL and esophageal manifestations and CeD might be further investigated. In addition, also the role of gluten-free diet in improving these conditions is largely unclear. Certainly, otorhinolaryngologists can play an important role in identifying people with unrecognized CeD and may help prevent its long-term complications. The aim of this narrative review is to analyze the latest evidence on the association between CeD and ORL and esophageal manifestations.

Background: Care workers in nursing homes often perform tasks that are rather related to organizational or management activities than 'direct patient care'. 'Indirect care activities', such as documentation or other administrative tasks are often considered by care workers as a burden, as they increase overall workload and keep them away from caring for residents. So far, there is little investigation into what kind of administrative tasks are being performed in nursing homes, by which type of care workers, and to which extent, nor how administrative burden is associated with care workers' outcomes., Purpose: The objective of this study was to describe care workers' administrative burden in Swiss nursing homes and to explore the association with four care worker outcomes (i.e., job dissatisfaction, emotional exhaustion, intention to leave the current job and the profession)., Methods: This multicenter cross-sectional study used survey data from the Swiss Nursing Homes Human Resources Project 2018. It included a convenience sample of 118 nursing homes and 2'207 care workers (i.e., registered nurses, licensed practical nurses) from Switzerland's German- and French-speaking regions. Care workers completed questionnaires assessing the administrative tasks and burden, staffing and resource adequacy, leadership ability, implicit rationing of nursing care and care worker characteristics and outcomes. For the analysis, we applied generalized linear mixed models, including individual-level nurse survey data and data on unit and facility characteristics., Results: Overall, 73.9% (n = 1'561) of care workers felt strongly or rather strongly burdened, with one third (36.6%, n = 787) reporting to spend 2 h or more during a "normal" day performing administrative tasks. Ratings for administrative burden ranged from 42.6% (n = 884; ordering supplies and managing stocks) to 75.3% (n = 1'621; filling out the resident's health record). One out of four care workers (25.5%, n = 561) intended to leave the profession, whereby care workers reporting higher administrative task burden (OR = 1.24; 95%CI: 1.02-1.50) were more likely to intend to leave the profession., Conclusion: This study provides first insights on care workers' administrative burden in nursing homes. By limiting care workers' burdensome administrative tasks and/or shifting such tasks from higher to lower educated care workers or administrative personnel when appropriate, nursing home managers could reduce care workers' workload and improve their job satisfaction and retention in the profession., (© 2023. The Author(s).)

This review summarizes recent investigations on dental manifestations in celiac disease. Particular attention is paid to delayed dental eruption and maturity, dental enamel defects, molar incisor hypomineralization, dental caries, dental plaque, and periodontitis. Most studies confirmed a higher frequency of delayed dental eruption and maturation in children and dental enamel defects in children and adults with celiac disease compared to healthy individuals. The malabsorption of various micronutrients, especially calcium and vitamin D, as well as immunity, is considered the main cause of these conditions. An early diagnosis of celiac disease and introducing a gluten-free diet might prevent the development of these conditions. Otherwise, the damage has already been established, and it is irreversible. Dentists can play an important role in identifying people who may have unrecognized celiac disease and may help prevent its progress and long-term complications. Investigations on dental caries, plaque, and periodontitis in celiac disease are rare and inconsistent; these complaints need further examination.

This study aimed to evaluate the validity and reliability of the German and Italian versions of the Interprofessional Collaboration Scale (IPC scale) by applying a Rasch analysis. Data were gathered from 1182 nurses participating in a cross-sectional study in northern Italy. The scale demonstrated good reliability (Cronbach's alpha = 0.92). Item polarity of all 13 items was positive, indicating good construct validity. However, revising one item would further improve the validity of the scale. Item stability was confirmed for work experience, workplace, age range, the language version, and gender. The analysis, applying a non-classical test theory, confirmed that the IPC scale is a valid and reliable instrument to measure interprofessional collaboration between nurses and physicians.

Celiac disease (CD) is a chronic gluten-responsive immune mediated enteropathy and is treated with a gluten-free diet (GFD). However, a strict diet for life is not easy due to the ubiquitous nature of gluten. This review aims at examining available evidence on the degree of adherence to a GFD, the methods to assess it, and the barriers to its implementation. The methods for monitoring the adherence to a GFD are comprised of a dietary questionnaire, celiac serology, or clinical symptoms; however, none of these methods generate either a direct or an accurate measure of dietary adherence. A promising advancement is the development of tests that measure gluten immunogenic peptides in stools and urine. Causes of adherence/non-adherence to a GFD are numerous and multifactorial. Inadvertent dietary non-adherence is more frequent than intentional non-adherence. Cross-contamination of gluten-free products with gluten is a major cause of inadvertent non-adherence, while the limited availability, high costs, and poor quality of certified gluten-free products are responsible for intentionally breaking a GFD. Therefore, several studies in the last decade have indicated that many patients with CD who follow a GFD still have difficulty controlling their diet and, therefore, regularly consume enough gluten to trigger symptoms and damage the small intestine.

A gluten-free diet (GFD) is currently the only effective treatment for celiac disease (CD); an individual's daily intake of gluten should not exceed 10 mg. However, it is difficult to maintain a strict oral diet for life and at least one-third of patients with CD are exposed to gluten, despite their best efforts at dietary modifications. It has been demonstrated that both natural and certified gluten-free foods can be heavily contaminated with gluten well above the commonly accepted threshold of 20 mg/kg. Moreover, meals from food services such as restaurants, workplaces, and schools remain a significant risk for inadvertent gluten exposure. Other possible sources of gluten are non-certified oat products, numerous composite foods, medications, and cosmetics that unexpectedly contain "hidden" vital gluten, a proteinaceous by-product of wheat starch production. A number of immunochemical assays are commercially available worldwide to detect gluten. Each method has specific features, such as format, sample extraction buffers, extraction time and temperature, characteristics of the antibodies, recognition epitope, and the reference material used for calibration. Due to these differences and a lack of official reference material, the results of gluten quantitation may deviate systematically. In conclusion, incorrect gluten quantitation, improper product labeling, and poor consumer awareness, which results in the inadvertent intake of relatively high amounts of gluten, can be factors that compromise the health of patients with CD.

Interprofessional working relationships can influence the quality of collaborative practices, with consequences for patient safety outcomes. This article reports findings of an adapted relationship scale comprising six different relationship types, ranging from hostile to collegial, between seven health-care professions: physicians, nurses, dieticians, occupational therapists, physiotherapists, speech therapists, and psychologists. Survey data were gathered within amixed-method study aiming to explore the current status quo of interprofessional collaboration in aHealth Trust, located in Northern Italy. An online questionnaire was completed by 2,238 health professionals achieving aresponse rate of 44%. The working relationship element was answered by 1,897 respondents. The results of the survey are used as abasis for recommending strategies for advancing interprofessional collaboration in the Trust. Descriptive statistics were used to examine relationship-types, with frequency of occurrence considered. The non-parametric Mann Whitney and Kruskal Wallis tests were used to explore relationship differences among groups defined by sociodemographic variables. Participants reported overall positive relationships with other health professions. We noted variability in the occurrence of different relationship-types amongst the health professions. In particular, the six professions viewed their relationships with doctors more negatively than physicians who reported amore positive perception of their relationships with the six professions.

The aim of our study was to evaluate the disposition of individuals with type 2 diabetes mellitus (DM2) toward changing their nutritional and physical activity habits and associated factors-particularly their perceptions about interacting and communicating with four health professions. Working with a local patients' association, we invited 364 individuals with DM2, all at least 18 years old, to complete a paper-based survey with questions addressing their experiences of interacting and communicating with general practitioners, nurses, dieticians and diabetologists and about their readiness to change targeted habits, their health literacy and their clinical status. Of the 109 questionnaires collected, 100 were eligible for descriptive and inferential statistical analysis. Regarding nutritional habits, the highest percentage of participants were at the maintenance stage (26%), whereas regarding physical activity habits the highest percentage of participants were at the preparation stage (31%). Significant differences between the habits emerged for four of the five stages and for two psychological processes. The precontemplation stage was most associated with communication-related variables, whereas the maintenance stage was associated with higher health literacy for both habits, and waist-to-height ratio was associated with several stages of change and psychological processes for physical activity habits. Considering aggregated stages (i.e., active or passive stage), significant differences were observed for all psychological processes except readiness to change nutritional habits. Logistic regression analysis revealed associations of the active stage with higher self-efficacy and lower discrepancy processes for both habits. Nutritional habits were associated with normal HbA1c values and physical activity habits with high cholesterol. Understanding the combination of the stages of change and how they relate to psychological processes can afford meaningful insights into the potential internal and external communication skills of health professions and should be examined as possible elements for a patient evaluation model.

Wheat-based foods have been staple foods since about 10,000 years and constitute a major source of energy, dietary fiber, and micronutrients for the world population. The role of wheat in our diet, however, has recently been scrutinized by pseudoscientific books and media reports promoting the overall impression that wheat consumption makes people sick, stupid, fat, and addicted. Consequently, numerous consumers in Western countries have started to question their dietary habits related to wheat consumption and voluntarily decided to adopt a wheat-free diet without a medical diagnosis of any wheat-related disorder (WRD), such as celiac disease, wheat allergy, or non-celiac gluten sensitivity. The aim of this review is to achieve an objective judgment of the positive aspects of wheat consumption as well as adverse effects for individuals suffering from WRDs. The first part presents wheat constituents and their positive nutritional value, in particular, the consumption of products from whole-grain flours. The second part is focused on WRDs that affect predisposed individuals and can be treated with a gluten-free or -reduced diet. Based on all available scientific knowledge, wheat consumption is safe and healthy for the vast majority of people. There is no scientific evidence to support that the general population would benefit from a wheat-free diet., (Copyright © 2020 Wieser, Koehler and Scherf.)

Objectives: To explore whether a team-based learning strategy applied to an interprofessional course on basic science changes students' perception of communication and teamwork skills and attitudes as related to interprofessional learning., Methods: A mixed-methods approach was utilized. The participants were selected through an opportunity sample of 33 first-semester anatomy students from occupational therapy and orthoptics programs. Students completed an interprofessional questionnaire before and after the course. The data were analyzed descriptively. Fourteen students were selected randomly for group interviews. Qualitative data was interpreted using thematic analyses., Results: The pre-test scores for 'communication and teamwork skills' and 'interprofessional learning' were high with mean values of 26.58 and 34.24, respectively. The post-test scores were 27.30 and 34.27, respectively, indicating no relevant changes in students' perception and attitudes. Qualitative data suggested that team-based learning represents a valid strategy to encourage communication and teamwork skills but revealed a lack of interprofessional exchange during the course. Students reported that classroom activities must require the professional knowledge of all participating groups in order to prevent a negative attitudinal shift towards interprofessional education in the later years of their studies., Conclusions: Implementing team-based learning in basic sciences can encourage communication and teamwork among students. Mixed classes can help socialize students of different professional groups, although they carry a risk of a negative attitudinal shift towards interprofessional education. Whether, and in what ways, effective interprofessional exchange during the teaching of basic sciences can be achieved needs further investigation.

Introduction: Diabetes mellitus type 2 (DM2), one of the four most important chronic diseases worldwide, is generally considered to be preventable. However, it is not yet sufficiently clear whether an aligned collaboration between different health professions could facilitate behavioral changes to be made by patients with DM2 regarding their eating and physical activity habits., Objective: To explore if and how far in current outpatient care for 3 health-care professions it is an objective to collaborate with each other supporting patients with DM2 in changing their eating and physical activity habits., Methods: We conducted 18 qualitative problem centered interviews with selected family physicians, nurses, dieticians working in outpatient setting and patients with DM2, transcribed verbatim, and analyzed with qualitative content analysis., Results: Issues identified ranged from description and reflection of current health-care practice, strategies, and hindrances to cope with changes of eating and physical activity behaviors as well as for health-care practice regarding interprofessional collaboration and patient-centered care up to considerations about collaboration and patient centricity (for health professionals and patients to achieve goals) and changes and ideas of "ideal care practice"., Discussion: The included professional groups work predominantly for themselves. Collaboration currently only takes place when individually triggered and neither structured nor organized., Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2020.)

This article presents quantitative findings from a mixed method study that aimed to explore the status quo of interprofessional collaboration (IPC) in a Health Trust, located in a trilingual region in Northern Italy. The survey targeted seven health professions (physicians, nurses, dieticians, occupational therapists, physiotherapists, speech therapists, and psychologists). The survey was distributed online to more than 5,000 health professionals and completed by 2,238. This paper presents results on the frequency of collaboration as well as data from a multiple-group measurement scale for assessing IPC. Descriptive statistics were calculated for continuous variables while categorical data were analysed as counts and percentages. Pearson's Chi-square test and Fisher's exact test were calculated while Mann Whitney and Kruskal Wallis tests were applied to analyse statistical differences in IPC between groups according to sociodemographic variables. In general, our survey data showed that participants, perceived IPC in a positive way, even if the analysis indicated heterogeneity in the level of collaboration expressed. We also found that not all professions had an opportunity to collaborate with others. In addition, we found evidence to suggest that the way health care is organised impacted on perceptions of collaboration between the health professionals in this study. This study provides an initial insight into the perceived levels of IPC within a North Italian context. As such, it offers an account of the strengths and weaknesses of IPC from seven different professional groups based in this region of Europe.

Female bats of temperate zones often communally rear their young, which creates ideal conditions for naive juveniles to find or learn about resources via informed adults. However, studying social information transfer in elusive and small-bodied animals in the wild is difficult with traditional tracking techniques. We used a novel 'next-generation' proximity sensor system (BATS) to investigate if and how juvenile bats use social information in acquiring access to two crucial resources: suitable roosts and food patches. By tracking juvenile-adult associations during roost switching and foraging, we found evidence for mother-to-offspring information transfer while switching roosts but not during foraging. Spatial and temporal patterns of encounters suggested that mothers guided juveniles between the juvenile and the target roost. This roost-switching behaviour provides evidence for maternal guidance in bats, a form of maternal care that has long been assumed, but never documented. We did not find evidence that mothers guide the offspring to foraging sites. Foraging bats reported brief infrequent meetings with other tagged bats that were best explained by local enhancement. Our study illustrates how this recent advance in automated biologging provides researchers with new insights into longstanding questions in behavioural biology.

Gluten is the trigger for celiac disease (CD), non-celiac gluten/wheat sensitivity (NCGS), and wheat allergy. An oral food challenge is often needed for diagnosis, but there are no standardized gluten challenge materials with known composition available. To fill this gap, two materials, commercially available gluten and a food-grade gluten hydrolysate (pepgluten), were extensively characterized. Pepgluten was prepared from gluten by incubation with a pepsin dietary supplement and acetic acid at 37 °C for 120 min. The components of pepgluten were crude protein (707 mg/g), starch (104 mg/g), water (59 mg/g), fat (47 mg/g), dietary fiber (41 mg/g) and ash (11 mg/g). The protein/peptide fraction of pepgluten (1 g) contained equivalents derived from 369 mg gliadins and 196 mg glutenins, resulting in 565 mg total gluten equivalents, 25 mg albumins/globulins, 22 mg α-amylase/trypsin inhibitors and 48 mg pepsin capsule proteins. The slightly acidic, dough-like smell and bitter taste of pepgluten could be completely camouflaged in multivitamin juice with bitter lemon, grapefruit juice, or vegetable and fruit smoothies. Thus, pepgluten met the criteria for placebo-controlled challenges (active and placebo materials are identical regarding appearance, taste, smell, and texture) and is appropriate as a standard preparation for the oral food challenge and clinical investigations to study wheat hypersensitivities., Competing Interests: H.W. is inventor of the utility model DE 202017005555 U1. K.A.S. declares no conflict of interest.

Celiac disease (CD), a chronic enteropathy of the small intestine caused by ingestion of gluten, is one of the most prevalent food hypersensitivities worldwide. The essential treatment is a strict lifelong gluten-free diet based on the avoidance of gluten-containing products from wheat, rye, barley and, in rare cases, oats. Products made from naturally gluten-free raw materials often have inferior nutritional, textural and sensory properties compared to the corresponding gluten-containing products. Therefore, the incorporation of wheat, rye and barley flours after efficient removal of the harmful component gluten into gluten-free products would be beneficial. Gluten modification resulting in decreased CD-immunoreactivity may be achieved via the formation of crosslinks using microbial transglutaminase. To effectively eliminate CD-immunoreactivity, plant, fungal, bacterial, animal or engineered peptidases are capable of degrading gluten proteins and peptides into harmless fragments. The application of peptidases from germinated cereal grains, fungal peptidases and/or lactic acid bacteria during food processing yielded high-quality sourdough wheat breads, pasta, wheat starch and bran, rye products and beer, all with gluten contents below the Codex Alimentarius threshold of 20mg/kg for gluten-free products. As with all gluten-free products, the legislative compliance of such treated materials needs to be monitored closely. Provided that all safety requirements are met, gluten-containing raw materials treated in an adequate way to remove CD-active gluten fragments may be used together with naturally gluten-free ingredients to create an extended choice of high-quality gluten-free products., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This article presents a study that aimed to validate a translation of a multiple-group measurement scale for interprofessional collaboration (IPC). We used survey data gathered over a three month period as part of a mixed methods study that explored the nature of IPC in Northern Italy. Following a translation from English into Italian and German the survey was distributed online to over 5,000 health professionals (dieticians, nurses, occupational therapists, physicians, physiotherapists, speech therapists and psychologists) based in one regional health trust. In total, 2,238 different health professions completed the survey. Based on the original scale, three principal components were extracted and confirmed as relevant factors for IPC (communication, accommodation and isolation). A confirmatory analysis (3-factor model) was applied to the data of physicians and nurses by language group. In conclusion, the validation of the German and Italian IPC scale has provided an instrument of acceptable reliability and validity for the assessment of IPC involving physicians and nurses.

Coeliac disease (CD) is triggered by the ingestion of gluten proteins from wheat, rye, and barley. The 33-mer peptide from α2-gliadin has frequently been described as the most important CD-immunogenic sequence within gluten. However, from more than 890 published amino acid sequences of α-gliadins, only 19 sequences contain the 33-mer. In order to make a precise assessment of the importance of the 33-mer, it is necessary to elucidate which wheat species and cultivars contain the peptide and at which concentrations. This paper presents the development of a stable isotope dilution assay followed by liquid chromatography tandem mass spectrometry to quantitate the 33-mer in flours of 23 hexaploid modern and 15 old common (bread) wheat as well as two spelt cultivars. All flours contained the 33-mer peptide at levels ranging from 91-603 μg/g flour. In contrast, the 33-mer was absent (

Purified wheat starch (WSt) is commonly used in gluten-free products for celiac disease (CD) patients. It is mostly well-tolerated, but doubts about its safety for CD patients persist. One reason may be that most ELISA kits primarily recognize the alcohol-soluble gliadin fraction of gluten, but insufficiently target the alcohol-insoluble glutenin fraction. To address this problem, a new sensitive method based on the sequential extraction of gliadins, glutenins, and gluten from WSt followed by gel-permeation high-performance liquid chromatography with fluorescence detection (GP-HPLC-FLD) was developed. It revealed that considerable amounts of glutenins were present in most WSt. The gluten contents quantitated by GP-HPLC-FLD as sum of gliadins and glutenins were higher than those by R5 ELISA (gluten as gliadin content multiplied by a factor of 2) in 19 out of 26 WSt. Despite its limited selectivity, GP-HPLC-FLD may be applied as confirmatory method to ELISA to quantitate gluten in WSt.

Objective: The purpose of this study was to investigate how students perceived their learning experience when combining traditional anatomy lectures with preparatory e-learning activities that consisted of fill-in-the-blank assignments, videos, and multiple-choice quizzes., Methods: A qualitative study was conducted to explore changes in study behaviour and perception of learning. Three group interviews with students were conducted and thematically analysed., Results: Data was categorized into four themes: 1. Approaching the course material, 2. Understanding the material, 3. Consolidating the material, and 4. Perceived learning outcome. Students appreciated the clear structure of the course, and reported that online activities encouraged them towards a first engagement with the material. They felt that they were more active during in-class sessions, described self-study before the end-of-term exam as easier, and believed that contents would remain in their memories for a longer time., Conclusions: By adjusting already existing resources, lectures can be combined fairly easily and cost-effectively with preparatory e-learning activities. The creation of online components promote well-structured courses, can help minimize 'student passivity' as a characteristic element of lectures, and can support students in distributing their studies throughout the term, thus suggesting enhanced learning. Further research work should be designed to confirm the afore-mentioned findings through objective measurements of student learning outcomes.

Wheat-dependent exercise-induced anaphylaxis (WDEIA) is a rare, but potentially severe food allergy exclusively occurring when wheat ingestion is accompanied by augmenting cofactors. It is clinically characterized by anaphylactic reactions ranging from urticaria and angioedema to dyspnoea, hypotension, collapse, and shock. WDEIA usually develops after ingestion of wheat products followed by physical exercise. Other cofactors are acetylsalicylic acid and other non-steroidal anti-inflammatory drugs, alcohol, and infections. The precise mechanisms of WDEIA remain unclear; exercise and other cofactors might increase gastrointestinal allergen permeability and osmolality, redistribute blood flow, or lower the threshold for IgE-mediated mast cell degranulation. Among wheat proteins, ω5-gliadin and high-molecular-weight glutenin subunits have been reported to be the major allergens. In some patients, WDEIA has been discussed to be caused by epicutaneous sensitization with hydrolysed wheat gluten included in cosmetics. Diagnosis is made based on the patient's history in combination with allergy skin testing, determination of wheat-specific IgE serum antibodies, basophil activation test, histamine release test, and/or exercise challenge test. Acute treatment includes application of adrenaline or antihistamines. The most reliable prophylaxis of WDEIA is a gluten-free diet. In less severe cases, a strict limitation of wheat ingestion before exercise and avoidance of other cofactors may be sufficient., (© 2015 John Wiley & Sons Ltd.)

Wholemeal flours from 62 spelt and 13 wheat cultivars were studied. The quantitative protein compositions of the Osborne fractions determined by reversed-phase high-performance liquid chromatography, showed that the chromatograms of the reduced gliadin fractions were most suitable for the distinction of spelt from wheat and for the classification of spelt. The patterns of the reduced spelt gliadins showed one to three markers that were not present in wheat. Based on these markers, spelt cultivars were classified into three groups ranging from 'typical spelt' to 'similar to common wheat'. Marker 1 was identified as ω1,2-gliadin and markers 2, 3a and 3b were identified as γ-gliadins by means of N-terminal sequence analysis and determination of the relative molecular mass by mass spectrometry. As glutenin-bound ω-gliadins were present in wheat and absent in spelt, this protein type may be used to detect and quantitate small amounts of wheat in spelt products., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Health care systems are facing continual reorganizations in response to scientific and technological innovations as well as financial constraints. Simultaneously the rising prevalence of chronic diseases call for a kind of health care organization in which interprofessional collaboration (IPC) functions on a high level. This article describes a research project that aims to generate an empirical account of the current state of IPC in the South Tyrolean Health Trust, located in a bilingual region in northern Italy. The study will employ a sequential mixed methods' design. A survey will be followed by individual and focus group interviews. An innovative aspect of the study is that it will include participants from six different health professions (dieticians, nurses, occupational therapists, physicians, physiotherapists, and speech therapists). Qualitative data analysis will draw on the negotiated order/structural ordering approach and on the social world/arena perspective. The results are expected to generate specific recommendations to improve IPC in the South Tyrolean Health Trust and inform further research.

Ingestion of wheat, barley, or rye triggers small intestinal inflammation in patients with celiac disease. Specifically, the storage proteins of these cereals (gluten) elicit an adaptive Th1-mediated immune response in individuals carrying HLA-DQ2 or HLA-DQ8 as major genetic predisposition. This well-defined role of adaptive immunity contrasts with an ill-defined component of innate immunity in celiac disease. We identify the α-amylase/trypsin inhibitors (ATIs) CM3 and 0.19, pest resistance molecules in wheat, as strong activators of innate immune responses in monocytes, macrophages, and dendritic cells. ATIs engage the TLR4-MD2-CD14 complex and lead to up-regulation of maturation markers and elicit release of proinflammatory cytokines in cells from celiac and nonceliac patients and in celiac patients' biopsies. Mice deficient in TLR4 or TLR4 signaling are protected from intestinal and systemic immune responses upon oral challenge with ATIs. These findings define cereal ATIs as novel contributors to celiac disease. Moreover, ATIs may fuel inflammation and immune reactions in other intestinal and nonintestinal immune disorders.

Disulfide bonds within gluten proteins play a key role in the breadmaking performance of wheat flour. In the present study, disulfide bonds of wheat gluten proteins were identified by using a new liquid chromatography-mass spectrometry (LC-MS) technique with alternating electron transfer dissociation (ETD)/collision-induced dissociation (CID). Wheat flour was partially hydrolyzed with thermolysin (pH 6.5, 37 °C, 16 h), and the digest was subjected to LC-MS with alternating ETD/CID fragmentation. Whereas CID provided peptide fragments with intact disulfide bonds, cleavage of disulfide bonds was preferred over peptide backbone fragmentations in ETD. The simultaneous observation of disulfide-linked and disulfide-cleaved peptide ions in the mass spectra not only provided distinct interpretation with high confidence but also simplified the conventional approach for determination of disulfide bonds, which often requires two separate experiments with and without chemical reduction. By application of the new method 14 cystine peptides were identified. Eight peptides confirmed previously established disulfide bonds within gluten proteins, and the other six cystine peptides were identified for the first time. One of the newly identified cystine peptides represented a "head-to-tail" cross-link between high molecular weight glutenin subunits. This type of cross-link, which has been postulated as an integral part of glutenin models published previously, has now been proven experimentally for the first time. From the six remaining cystine peptides interchain disulfide bonds between α-gliadins, γ-gliadins, and low molecular weight glutenin subunits were established.

Celiac disease (CD) is an inflammatory disease of the upper small intestine in genetically predisposed individuals caused by glutamine- and proline-rich peptides from cereal storage proteins (gluten) with a minimal length of nine amino acids. Such peptides are insufficiently degraded by gastrointestinal enzymes; they permeate the lymphatic tissue, are bound to celiac-specific, antigen-presenting cells, and stimulate intestinal T-cells. The typical clinical pattern is a flat small intestinal mucosa and malabsorption. Currently, the only therapy is a strict, lifelong gluten-free diet. Recent research has shown that gluten and gluten peptides can be degraded by prolyl endopeptidases from different sources. These peptidases can either be used to produce gluten-free foods from gluten-containing raw materials, or they have been suggested as an oral therapy for CD, in which dietary gluten is hydrolyzed by coingested peptidases already in the stomach, thus preventing CD-specific immune reactions in the small intestine. This would be an alternative for CD patients to the gluten-free diet. Furthermore, microbial transglutaminase could be used to detoxify gluten either by selectively modifying glutamine residues of intact gluten by transamidation with lysine methyl ester or by crosslinking gluten peptides in beverages via isopeptide bonds so that they can be removed by filtration.

The four-stranded G-quadruplex motif is a conformation frequently adopted by guanine-rich nucleic acids that plays an important role in biology, medicine, and nanotechnology. Although vibrational spectroscopy has been widely used to investigate nucleic acid structure, association of particular spectral features with the quadruplex structure has to date been ambiguous. In this work, experimental IR absorption and vibrational circular dichroism (VCD) spectra of the model quadruplex systems d(G)(8) and deoxyguanosine-5'-monophosphate (5'-dGMP) were analyzed using molecular dynamics (MD) and quantum-chemical modeling. The experimental spectra were unambiguously assigned to the quadruplex DNA arrangement, and several IR and VCD bands related to this structural motif were determined. Involvement of MD in the modeling was essential for realistic simulation of the spectra. The VCD signal was found to be more sensitive to dynamical structural variations than the IR signal. The combination of the spectroscopic techniques with multiscale simulations provides extended information about nucleic acid conformations and their dynamics.

Objective: Coeliac disease (CD) is a multisystemic autoimmune inflammation of the intestinal tract induced by wheat gluten and related cereals in human leucocyte antigen (HLA)-DQ2/8-positive individuals. The molecular mechanisms relevant to oral tolerance induction towards toxic cereals such as gliadin remain poorly understood. Enterocytes, which express predominantly HLA-DR proteins, are capable of processing, transcytosing and presenting food antigens from the intestinal lumen to T lymphocytes of the lamina propria., Methods: Epitope-specific monoclonal antigliadin antibodies are utilised to unravel the intraepithelial transport processes of gliadin peptides in human duodenal biopsy specimens from patients with CD and reconstitute the transepithelial and endocytic pathways of gliadin in intestinal epithelial HT29 cells., Results: The gliadin peptide AA 31-49 is segregated from the peptides AA 56-68 and AA 229-246 along the endosomal pathway. Thus, AA 31-49 bypasses HLA-DR-positive late endosomes in intestinal cells and in biopsy specimens of patients with untreated CD. Further, it is localised in early endosomes and consequently escapes antigen presentation at the basolateral membrane, unlike peptides AA 56-68 and AA 229-246 that reach HLA-DR-positive late endosomes. Strikingly, forms of gliadin peptide AA 31-49 conjugated to cholera toxin B are sorted into late endosomes of HT29 cells., Conclusions: Endocytic segregation of gliadin peptide AA 31-49 seems to be a constitutive process. It explains why this peptide cannot stimulate gluten-sensitive T cells. Presentation of gliadin peptides by HLA-DR proteins via late endosomes within enterocytes might induce a tolerogenic effect and constitutes a potentially promising therapeutic approach for induction of tolerance towards gliadin.

Celiac disease (CD) is a permanent gastrointestinal disorder characterized by the intolerance to a group of proteins called gluten present in wheat, rye, barley, and possibly oats. The only therapy is a strict lifelong gluten-free diet. The standard method for gluten determination in foods produced for CD patients is the R5-enzyme-linked immunosorbent assay (ELISA) as proposed by the recent Codex Alimentarius Draft Revised Standard. This test is based on the determination of prolamins, the alcohol-soluble proteins of gluten, and is available as a sandwich ELISA for intact proteins and as a competitive ELISA for gluten-derived peptides. While the suitability of the sandwich ELISA including a wheat prolamin (gliadin) reference for calibration has been shown by various studies and a ring test, the competitive ELISA still lacks a convenient reference for the quantitation of gluten peptides in fermented cereal foods (e.g., sourdough products, starch syrup, malt extracts, beer). Therefore, the aim of the present study was to prepare a suitable reference for the quantitation of partially hydrolyzed gluten in fermented wheat, rye, and barley products. The prolamin fractions from barley (hordein) and rye (secalin) were isolated from corresponding flours by means of a modified preparative Osborne fractionation. The prolamin fraction from wheat was obtained as reference gliadin from the Prolamin Working Group. The prolamin fractions were successively digested by pepsin and trypsin or pepsin and chymotrypsin procedures, which have been used for CD-specific toxicity tests on cereal storage proteins for many years. The protein/peptide content (N x 5.7) of the prolamin fractions and digests, which was the basis for the calculation of the gluten content by means of ELISA, varied between 67.1% and 96.0%. The prolamin fractions and enzymatic digests were then tested for their response in both sandwich and competitive assays. Intact prolamins responded similarly in both ELISA showing no important differences between the cereals. In the case of digested proteins, however, the sandwich ELISA was considerably less sensitive than the competitive ELISA. The former provided approximately 40% and the latter 70% of the signal intensity obtained with the intact prolamins. Thus, the combination of the competitive ELISA and the enzymatic digests of prolamin fractions as reference was considered to be an adequate system for the analysis of partially hydrolyzed gluten. The limit of detection using a peptic-tryptic hordein digest as reference was 2.3 microg prolamin equivalent per milliliter, and the limit of quantitation was 6.7 microg prolamin equivalent per milliliter. This system was applied for the determination of gluten equivalents in five commercial beverages based on fermented cereals.

Spring wheat (Triticum aestivum L. cv. TRISO) was grown for three consecutive seasons in a free-air carbon dioxide (CO(2)) enrichment (FACE) field experiment in order to examine the effects on crop yield and grain quality. CO(2) enrichment promoted aboveground biomass (+11.8%) and grain yield (+10.4%). However, adverse effects were predominantly observed on wholegrain quality characteristics. Although the thousand-grain weight remained unchanged, size distribution was significantly shifted towards smaller grains, which may directly relate to lower market value. Total grain protein concentration decreased significantly by 7.4% under elevated CO(2), and protein and amino acid composition were altered. Corresponding to the decline in grain protein concentration, CO(2) enrichment resulted in an overall decrease in amino acid concentrations, with greater reductions in non-essential than essential amino acids. Minerals such as potassium, molybdenum and lead increased, while manganese, iron, cadmium and silicon decreased, suggesting that adjustments of agricultural practices may be required to retain current grain quality standards. The concentration of fructose and fructan, as well as amounts per area of total and individual non-structural carbohydrates, except for starch, significantly increased in the grain. The same holds true for the amount of lipids. With regard to mixing and rheological properties of the flour, a significant increase in gluten resistance under elevated CO(2) was observed. CO(2) enrichment obviously affected grain quality characteristics that are important for consumer nutrition and health, and for industrial processing and marketing, which have to date received little attention.

Increasing prices for wheat products and fertilizers, as well as reduced sulfur (S) contributions from the atmosphere, call for an improvement of product quality and agricultural management. To detect the impact of a time-dependent S fertilization, the quantitative protein composition and the baking quality of two different wheat cultivars, Batis and Turkis, were evaluated. The glutathione concentration in grains serves as a reliable marker of the need for added S fertilizer. The quantitation of gliadins and glutenin subunits by reversed-phase high-performance liquid chromatography confirmed that S-rich proteins significantly increased with S fertilization, whereas the S-poor proteins significantly decreased. Proteome analysis by means of high-resolution protein profiles detected 55 and 37 proteins from Batis and Turkis changed by late S fertilization. A microscale baking test using wholemeal flour was implemented for the evaluation of baking quality, and late S fertilization was found to improve the composition of gluten proteins and baking quality.

A method for the simultaneous quantitation of total glutathione and total cysteine in wheat flour by a stable isotope dilution assay using high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) was developed. As internal standards, L-[(13)C3, (15)N]cysteine and L-gamma-glutamyl-L-[(13)C3, (15)N]cysteinyl-glycine were used. The method consisted of the extraction and reduction of flour with tris(2-carboxyethyl) phosphine after the addition of internal standards, protection of free thiol groups with iodoacetic acid, derivatization of free amino groups with dansyl chloride, and HPLC-MS/MS. The limits of detection and quantitation for glutathione were 0.75 nmol/g and 2.23 nmol/g flour, respectively. For cysteine, the limits of detection and quantitation were 0.72 nmol/g and 2.12 nmol/g flour, respectively. The developed method was found to be sensitive enough for quantitation of total glutathione and cysteine levels in wheat flour. This method was then utilized to investigate the effect of sulfur (S) deficiency on the amount of total glutathione and cysteine in flour. In S-deficient wheat, the concentrations of total glutathione and cysteine were proportional to the amount of S supplied during growth. The calculation of correlations revealed that GSH and Cys concentrations influenced the rheological dough properties and the baking performance at least as much as protein parameters. Thus, the low concentration of GSH and Cys in flour from S-deficient wheat had a similar effect on the technological properties as the altered composition of gluten proteins.

The continuing increase in atmospheric CO 2 concentration is predicted to enhance biomass production and to alter biochemical composition of plant tissues. In the present study, winter wheat ( Triticum aestivum L. cv. 'Batis') was grown under ambient air (BLOW, CO 2 concentration: 385 muL L (-1)) and free-air CO 2 enrichment (FACE, CO 2 concentration: 550 muL l (-1)) and two different nitrogen (N) fertilization levels (normal N supply: N100, 50% of normal N supply: N50). Mature kernels were milled into white flour and analyzed for the contents of crude protein, Osborne fractions, single gluten protein types and glutenin macropolymer. Elevated CO 2 caused significant reductions in crude protein and all protein fractions and types ( p < 0.001) except albumins and globulins. Effects were more pronounced in wheat samples supplied with normal amounts of N fertilizer. Crude protein was reduced by 14% (N100) and 9% (N50), gliadins by 20% and 13%, glutenins by 15% and 15% and glutenin macropolymer by 19% and 16%, respectively. Within gliadins, omega5-gliadins (-35/-22%) and omega1,2-gliadins (-27/-14%) were more affected than alpha-gliadins (-21/-13%) and gamma-gliadins (-16/-12%). Within glutenins, HMW subunits (-23/-18%) were more affected than LMW subunits (-12/-15%). According to these results, flour from high CO 2 grown grain will have a diminished baking quality. To our knowledge, these are the first results of elevated CO 2 concentrations impacts on wheat grain protein composition gained under relevant growing conditions at least for Central Europe.

Background: In celiac disease gluten, the disease-inducing toxic component in wheat, induces the secretion of autoantibodies which are targeted against transglutaminase 2 (TG2). These autoantibodies are produced in the small-intestinal mucosa, where they can be found deposited extracellularly below the epithelial basement membrane and around mucosal blood vessels. In addition, during gluten consumption these autoantibodies can also be detected in patients' serum but disappear from the circulation on a gluten-free diet. Interestingly, after adoption of a gluten-free diet the serum autoantibodies disappear from the circulation more rapidly than the small-intestinal mucosal autoantibody deposits. The toxicity of gluten and the secretion of the disease-specific autoantibodies have been widely studied in organ culture of small-intestinal biopsy samples, but results hitherto have been contradictory. Since the mucosal autoantibodies disappear slowly after a gluten-free diet, our aim was to establish whether autoantibody secretion to organ culture supernatants in treated celiac disease patient biopsies is related to the duration of the diet and further to the pre-existence of mucosal TG2-specific IgA deposits in the cultured biopsy samples., Results: In the organ culture system conducted with biopsies derived from treated celiac disease patients, gliadin induced secretion of autoantibodies to culture supernatants, reduced epithelial cell height and increased the density of lamina proprial CD25+ cells. However, these changes could be demonstrated only in biopsies from short-term treated celiac disease patients, where the small-intestinal mucosal TG2-specific IgA autoantibody deposits were still present. Furthermore, in these biopsies autoantibody secretion could be stimulated fully only after a 48-hour gliadin challenge., Conclusion: Our results show that studies focusing on the toxic effects of gliadin in the organ culture system should be carried out with biopsy samples from short-term treated celiac disease patients who are likely still to have mucosal IgA deposits present. In addition to providing an explanation for the discrepancies in previous publications, the present study also enables further validation of the organ culture method.

The cis-platin binding to the d(CCTGGTCC)*d(GGACCAGG) model DNA octamer was monitored with infrared absorption (IR) and vibrational circular dichroism (VCD) spectroscopies. The spectra were modeled with the aid of density functional computations and a Cartesian coordinate-based transfer of molecular property tensors from smaller DNA fragments. Because of the fragmentation, the tensors could be calculated with a higher precision. Environmental effects, such as the presence of the solvent or the cis-platin ligand, could be included in the modeling. The solvent was modeled by an explicit inclusion of hydrogen-bound water molecules, positions of which were estimated from a molecular dynamics simulation, or by the polarized continuum COSMO model. The B3LYP and BPW91 functionals used for the calculations of the spectral parameters were combined with the relativistic LANL2DZ platinum pseudo-potentials. The simulations reproduced the main IR and VCD DNA spectral features and explained most of the changes observed experimentally upon metal binding. The results confirmed that the influence of the ligand on DNA vibrational properties is quite complex; it originates in the geometry deformation and normal mode coupling pattern changes of the platinated octamer, as well as in local perturbations of the electronic structure and force field of the GC base pairs to which the platinum is bound. Many of the local effects could be accounted for by a point charge used in place of the metal in the GC complex.

Barley has for feeding purposes a shortage of essential amino acids, especially lysine, threonine, and methionine, and an excess of proline and glutamine. In the present study, we have introduced into barley an antisense construct against C-hordeins, the storage protein with the lowest nutritional quality. SDS-PAGE and reverse phase HPLC revealed a relative reduction in the amounts of C-hordeins and relative increases in the content of the other storage proteins. The five different lines analyzed had lower amounts of proline, glutamic acid/glutamine, and phenylalanine (up to 12%, 6%, and 9% reductions), while the lysine, threonine, and methionine content was increased with up to 16%, 13% and 11%. It is concluded that antisense mediated suppression of C-hordein synthesis may be a promising approach for improving the nutritional value of barley as a feed crop while at the same time reducing the environmental nitrogen load.

Wheat kernels of the cultivar 'Tommi' were germinated for up to 168 h at 15, 20, 25, or 30 degrees C. Samples were taken at different stages of germination and were analyzed for the quantitative protein composition using an extraction/HPLC method, for folate vitamers using a stable isotope dilution assay, and for soluble, insoluble, and total dietary fiber using a gravimetric method. Gluten proteins were substantially degraded during germination. During the first stages of germination the degradation of glutenins was predominant, whereas longer germination times were required to degrade gliadins. The optimal temperature for gliadin degradation was 20 degrees C, and that for glutenin degradation was 25 degrees C. Omega5- and omega1,2-gliadins were less sensitive to proteolytic degradation than alpha- and gamma-gliadins, and LMW subunits of glutenin were less sensitive than HMW subunits. During germination a time- and temperature-dependent increase of total folate occurred. A maximum 3.6-fold concentration was obtained after 102 h of germination at 20 and 25 degrees C including 5-methyltetrafolate as the major vitamer. The concentration of dietary fiber remained constant or decreased during the first 96 h of germination. Prolonged germination times of up to 168 h led to a substantial increase of total dietary fiber and to a strong increase of the soluble dietary fiber by a factor of 3, whereas the insoluble fiber decreased by 50%.

Sulfur (S) fertilization has been long-known to influence the amounts of total free amino acids in plants. To determine the impact of S deficiency in wheat on the concentration of, in particular, free asparagine, the spring wheat cultivar 'Star' was grown in a laboratory scale (5 L pot) at five different levels of S fertilization. After maturity, the kernels were milled into white flours (1-5) and analyzed for their contents of total S and total nitrogen as well as for free amino acids and glucose, fructose, maltose, and sucrose. Extremely high concentrations of free asparagine (Asn; 3.9-5.7 g/kg) were determined in flours 1 and 2 (30 and 60 mg of S), whereas much lower amounts (0.03-0.4 g/kg) were present in flours grown at higher S levels. The amounts of the reducing carbohydrates were, however, scarcely affected by S fertilization. In agreement with the high amount of Asn in flours 1 and 2, heating of both flours led to the generation of very high amounts of acrylamide (1.7-3.1 mg/kg) as well as of 3-aminopropionamide (40-76 mg/kg). Similar concentrations were measured in crispbread prepared from both flours. Application of rheological measurements on doughs prepared from each flour and a determination of the loaf volume of bread baked therefrom clearly indicated that flours 1 and 2 would be excluded from commercial bread processing due to their poor technological properties. Two commercial flours showed relatively low concentrations of acrylamide after a thermal treatment.

Gluten proteins play a key role in determining the unique baking quality of wheat by conferring water absorption capacity, cohesivity, viscosity and elasticity on dough. Gluten proteins can be divided into two main fractions according to their solubility in aqueous alcohols: the soluble gliadins and the insoluble glutenins. Both fractions consist of numerous, partially closely related protein components characterized by high glutamine and proline contents. Gliadins are mainly monomeric proteins with molecular weights (MWs) around 28,000-55,000 and can be classified according to their different primary structures into the alpha/beta-, gamma- and omega-type. Disulphide bonds are either absent or present as intrachain crosslinks. The glutenin fraction comprises aggregated proteins linked by interchain disulphide bonds; they have a varying size ranging from about 500,000 to more than 10 million. After reduction of disulphide bonds, the resulting glutenin subunits show a solubility in aqueous alcohols similar to gliadins. Based on primary structure, glutenin subunits have been divided into the high-molecular-weight (HMW) subunits (MW=67,000-88,000) and low-molecular-weight (LMW) subunits (MW=32,000-35,000). Each gluten protein type consists or two or three different structural domains; one of them contains unique repetitive sequences rich in glutamine and proline. Native glutenins are composed of a backbone formed by HMW subunit polymers and of LMW subunit polymers branched off from HMW subunits. Non-covalent bonds such as hydrogen bonds, ionic bonds and hydrophobic bonds are important for the aggregation of gliadins and glutenins and implicate structure and physical properties of dough.

The interactions were studied of DNA with the nonhistone chromatin protein HMGB1 and histone H1 in the presence of manganese(II) ions at different protein to DNA and manganese to DNA phosphate ratios by using absorption and optical activity spectroscopy in the electronic [ultraviolet (UV) and electronic circular dichroism ECD)] and vibrational [infrared (IR) and vibrational circular dichroism (VCD)] regions. In the presence of Mn2+, the protein-DNA interactions differ from those without the ions and cause prominent DNA compaction and formation of large intermolecular complexes. At the same time, the presence of HMGB1 and H1 also changed the mode of interaction of Mn2+ with DNA, which now takes place mostly in the major groove of DNA involving N7(G), whereas interactions between Mn2+ and DNA phosphate groups are weakened by histone molecules. Considerable interactions were also detected of Mn2+ ions with aspartic and glutamic amino acid residues of the proteins., (Copyright 2006 Wiley Periodicals, Inc.)

The vibrational circular dichroism (VCD) and infrared absorption (IR) spectra of the mismatched octamer oligonucleotides d(CGTGCGCG)(2) (CGT) and d(CGCGTGCG)(2) (CGC) and their complexes with the antitumor drug daunomycin were measured in D(2)O, interpreted, and compared to the octamer d(CGCGCGCG)(2) (CG). The IR spectra of the mismatched octamers in the carbonyl-stretching region are similar to those of the parent CG, whereas the VCD spectra differ in several respects between each other. The main VCD feature due to carbonyl stretching is informative for the mismatches and CG. Vibrational modes in the sugar-phosphate region remain essentially unchanged especially for PO(2) (-) symmetric stretching. Differences between the free and complexed mismatch octamers occurred mainly in the carbonyl-stretching region (1,700-1,600 cm(-1)). The absorption intensity of the C==O peak of G is more prominent for CGC than CGT and resembles CG in this respect. The detailed composition of this doublet is clearly visible, indicating the geometric rearrangement of the base pairs in the presence of the mismatch and upon forming the daunomycin complex.

Background: In coeliac disease, wheat, barley and rye are traditionally excluded in the gluten-free diet. However, few studies have examined the small intestinal immune response to barley and rye., Aim: To investigate the immunogenicity of barley and rye prolamins (hordein and secalin respectively) in comparison with wheat gliadin., Methods: Duodenal biopsies from 22 coeliac patients and 23 disease controls were cultured for 4 h with gliadin, hordein or secalin and compared with culture medium alone. Proinflammatory cytokines, interferon-gamma and interleukin-2, were quantified by TaqMan polymerase chain reaction and enzyme-linked immunosorbent assay., Results: Hordein caused the greatest increase in interferon-gamma mRNA in coeliac patients (median: 3.3-fold) in comparison with control subjects (median: 0.28-fold, P < 0.085). Secalin and gliadin induced similar levels of interferon-gamma mRNA with median fold-changes of 3.4 and 2.8, respectively, in coeliac patients in comparison with 1.6- and 1.1-fold increases in control subjects (P < 0.294 and P < 0.105, respectively). The median fold-changes for interleukin-2 mRNA did not differ between coeliac patients and controls. Cytokine protein was not upregulated., Conclusion: The findings of this study provide evidence that barley and rye cause immune activation in the mucosa of coeliac patients and support the practice that barley and rye should be excluded from the gluten-free diet.

Objectives: The ability of the gliadin fraction of wheat gluten to exacerbate coeliac disease is well documented. We investigated the possible toxicity of high molecular weight glutenin subunits (HMW-GS) in coeliac disease in vitro using gluten-sensitive T cells, and in vivo with challenge studies in patient volunteers., Methods: A mixture of four HMW-GS was chemically separated from wheat flour and checked for purity by HPLC, SDS-PAGE and ELISA. T-cell lines, grown up from small intestinal biopsies from coeliac patients (n=17), were tested for their reactivity to HMW-GS. Adults with coeliac disease and who were on a gluten-free diet (n=3) underwent in-vivo challenges with HMW-GS. Duodenal biopsies, taken prior to the challenge and at intervals up to 6 h afterwards, were assessed for morphology, intra-epithelial lymphocyte count, and interleukin 15 (IL-15) expression, by immunohistochemistry., Results: T-cell lines from 11 of 17 patients were stimulated by HMW-GS. There was a significant change in small intestinal morphology 4 h after commencing infusions with HMW-GS in all three subjects. For example villus height to crypt depth ratios were reduced in the three patients from 3.0+/-0.5 to 1.29+/-0.2, 2.53+/-0.7 to 0.81+/-0.6 and 3.0+/-0.7 to 1.85+/-0.3, P<0.0001 in all cases. There was increased expression of IL-15 in the small intestine from 2 h after the HMW-GS challenges., Conclusion: Mixed HMW-GS stimulate T-cell lines from some coeliac patients and exacerbate coeliac disease in vivo, inducing expression, within 2 h, of IL-15. This suggests an innate immune response to these proteins.

Background and Aims: Intestinal inflammation in coeliac disease is driven by the gluten fraction of wheat proteins. Deamidation or cross linking of gluten peptides by tissue transglutaminase (tTG), the coeliac disease autoantigen, creates potent T cell stimulatory peptides. Therefore, our aim was to identify the reaction patterns of gluten peptides, intestinal extracellular matrix proteins, and tTG., Methods: tTG activity was analysed by incorporation of monodansyl cadaverine into gliadins. Fluorescence labelled tTG reactive short gliadin peptides were used to demonstrate their deamidation and explore their cross linking patterns with tTG itself or extracellular matrix proteins. Patient sera and controls were checked for autoantibodies to matrix proteins., Results: Gliadins alpha1-alpha11, gamma1-gamma6, omega1-omega3, and omega5 were substrates for tTG. tTG catalysed the cross linking of gliadin peptides with interstitial collagen types I, III, and VI. Coeliac patients showed increased antibody titres against the collagens I, III, V, and VI., Conclusions: tTG formed high molecular weight complexes with all tested gliadins. As all tested gliadins were substrates for tTG, the tTG catalysed modifications were not restricted to single gliadin types and epitopes. Furthermore, haptenisation and long term immobilisation of gliadin peptides by tTG catalysed binding to abundant extracellular matrix proteins could be instrumental in the perpetuation of intestinal inflammation and some associated autoimmune diseases in coeliac disease.

Celiac disease is caused by sensitivity to wheat gluten in genetically susceptible individuals. The etiological role of the other wheat-related cereals, barley, rye, and oats, is still debated. In order to investigate this issue further, in this study we examined the immune response of celiac mucosal T cell lines to fractions from all four cereals. Cell stimulation was assessed by measuring proliferation (employing (3)H-thymidine incorporation) or cytokine (IL-2, IFN-gamma) production. All five T cell lines demonstrated immunoreactivity to protein fractions from the four related cereals. In some cell lines, reactivity to wheat, barley, and rye was only evident when these cereal fractions had been pretreated with tissue transglutaminase. This study confirms the similar T cell antigenic reactivity of these four related cereals and has implications for their exclusion in the gluten-free diet. However, despite oats stimulation of T cell lines, this cereal does not activate a mucosal lesion in most celiac patients.