Absolute and relative quantitation of amylase/trypsin-inhibitors by LC-MS/MS from wheat lines obtained by CRISPR-Cas9 and RNAi

Quantitation of wheat proteins is still a challenge, especially regarding amylase/trypsin-inhibitors (ATIs). A selection of ATIs was silenced in the common wheat cultivar Bobwhite and durum wheat cultivar Svevo by RNAi and gene editing, respectively, in order to reduce the amounts of ATIs. The controls and silenced lines were analyzed after digestion to peptides by LC-MS/MS with different approaches to evaluate changes in composition of ATIs. First, a targeted method with stable isotope dilution assay (SIDA) using labeled peptides as internal standards was applied. Additionally, four different approaches for relative quantitation were conducted, in detail, iTRAQ labeled and label free quantitation (LFQ) combined with data dependent acquisition (DDA) and data independent acquisition (DIA). Quantitation was performed manually (Skyline and MASCOT) and with different proteomics software tools (PLGS, MaxQuant, and PEAKS X Pro). To characterize the wheat proteins on protein level, complementary techniques as high-performance liquid chromatography (HPLC) and gel electrophoresis were performed. The targeted approach with SIDA was able to quantitate all ATIs, even at low levels, but an optimized extraction is necessary. The labeled iTRAQ approach revealed an indistinct performance. LFQ with low resolution equipment (IonTrap) showed similar results for major ATIs, but low abundance ATIs as CM1, were not detectable. DDA measurements with an Orbitrap system and evaluation using MaxQuant showed that the relative quantitation was dependent on the wheat species. The combination of manual curation of the MaxQuant search with Skyline revealed a very good performance. The DIA approach with analytical flow found similar results compared to absolute quantitation except for some minor ATIs, which were not detected. Comparison of applied methods revealed that peptide selection is a crucial step for protein quantitation. Wheat proteomics faces challenges due to the high genetic complexity, the close relationship to other cereals and the incomplete, redundant protein database requiring sensitive, precise and accurate LC-MS/MS methods

MOTIVATional intErviewing to Improve Self-Care in Heart Failure Patients (MOTIVATE-HF): Study Protocol of a Three-Arm Multicenter Randomized Controlled Trial

Aims Heart Failure (HF) self-care improves patient outcomes but trials designed to improve HF self-care have shown inconsistent results. Interventions may be more effective in improving self-care if they mobilize support from providers, promote self-efficacy, increase understanding of HF, increase the family involvement, and are individualized. All of these elements are emphasized in motivational interviewing (MI); few trials have been conducted using MI in HF patients and rarely have caregivers been involved in MI interventions. The aim of this study will be to evaluate if MI improves self-care maintenance in HF patients, and to determine if MI improves the following secondary outcomes: a) in HF patients: self-care management, self-care confidence, symptom perception, quality of life, anxiety/depression, cognition, sleep quality, mutuality with caregiver, hospitalizations, use of emergency services, and mortality; b) in caregivers: caregiver contribution to self-care, quality of life, anxiety/depression, sleep, mutuality with patient, preparedness, and social support. Methods A three-arm randomized controlled trial will be conducted in a sample of 240 HF patients and caregivers. Patients and caregivers will be randomized to the following arms: 1) MI intervention to patients only; 2) MI intervention to patients and caregivers; 3) standard of care to patients and caregivers. The primary outcome will be measured in patients 3 months after enrollment. Primary and secondary outcomes also will be evaluated 6, 9 and 12 months after enrollment. Conclusion This study will contribute to understand if MI provided to patients and caregivers can improve self-care. Because HF is rising in prevalence, findings can be useful to reduce the burden of the disease

Invasive pleural malignant mesothelioma with rib destruction and concurrent osteosarcoma in a dog

A 7-year-old Dachshund was clinically examined because of a 10-day history of lameness in the left hind limb. On the basis of radiological and cytological findings, an osteosarcoma of the left acetabular region was suspected. The dog underwent a hemipelvectomy and osteosarcoma was diagnosed by subsequent histopathological examination. An immovable subcutaneous mass was noted on the left chest wall during the physical examination and non-septic neutrophilic inflammation was diagnosed by cytology. Forty days later, the dog showed signs of respiratory distress with an in-diameter increase of the subcutaneous mass up to 4 cm. Thoracic radiography and ultrasonography revealed pleural effusion and a lytic process in the fourth left rib. Furthermore, ultrasound examination revealed a mixed echogenic mobile structure with a diameter of around 2 cm floating within the pleural fluid of the left hemithorax close to the pericardium. The dog underwent surgery for an en bloc resection of the subcutaneous mass together with the fourth rib and the parietal pleura. Moreover, the left altered lung lobe, corresponding to the mobile structure detected by ultrasound, was removed. Based on cytological, histopathological, and immunohistochemical examinations, an invasive epithelioid pleural malignant mesothelioma was diagnosed

Psychological Health Status of Psychiatric Patients Living in Treatment Communities before and during the COVID-19 Lockdown: A Brief Report

Many studies investigated the psychological impact of lockdown measures on the general population, while few studies focused on the psychiatric population. This study aimed to investigate the role of therapeutic communities in the management and containment of symptoms of patients with psychosis living in psychiatric residential facilities. Data were collected at two different points: November 2019 (Coronavirus disease 19 had not yet spread) and April 2020 (during the lockdown in Italy). Twenty-two study participants were recruited from three residential accredited psychiatric facilities. During lockdown, the patients showed a small increase in symptomatology in terms of emotional isolation. In addition, it was been observed significant differences in certain functional areas of the behavior, measured as lower inclination towards violent behaviors during lockdown, and higher scores in substance abuse and medical impairment. The lockdown condition could represent a form of containment; daily routines, along with adequate social support, are important aspects of the stability and the level of behavioral functioning of psychiatric patients. Social support and continuity of care offered by psychiatric communities can be an effective safeguard against the psychological impact of the COVID-19 epidemic

Wheat amylase/trypsin inhibitors (ATIs) : occurrence, function and health aspects

Amylase/trypsin inhibitors (ATIs) are widely consumed in cereal-based foods and have been implicated in adverse reactions to wheat exposure, such as respiratory and food allergy, and intestinal responses associated with coeliac disease and non-coeliac wheat sensitivity. ATIs occur in multiple isoforms which differ in the amounts present in different types of wheat (including ancient and modern ones). Measuring ATIs and their isoforms is an analytical challenge as is their isolation for use in studies addressing their potential effects on the human body. ATI isoforms differ in their spectrum of bioactive effects in the human gastrointestinal (GI), which may include enzyme inhibition, inflammation and immune responses and of which much is not known. Similarly, although modifications during food processing (exposure to heat, moisture, salt, acid, fermentation) may affect their structure and activity as shown in vitro, it is important to relate these changes to effects that may present in the GI tract. Finally, much of our knowledge of their potential biological effects is based on studies in vitro and in animal models. Validation by human studies using processed foods as commonly consumed is warranted. We conclude that more detailed understanding of these factors may allow the effects of ATIs on human health to be better understood and when possible, to be ameliorated, for example by innovative food processing. We therefore review in short our current knowledge of these proteins, focusing on features which relate to their biological activity and identifying gaps in our knowledge and research priorities.Peer reviewe

Invasive pleural malignant mesothelioma with rib destruction and concurrent osteosarcoma in a dog

A 7-year-old Dachshund was clinically examined because of a 10-day history of lameness in the left hind limb. On the basis of radiological and cytological findings, an osteosarcoma of the left acetabular region was suspected. The dog underwent a hemipelvectomy and osteosarcoma was diagnosed by subsequent histopathological examination. An immovable subcutaneous mass was noted on the left chest wall during the physical examination and non-septic neutrophilic inflammation was diagnosed by cytology. Forty days later, the dog showed signs of respiratory distress with an in-diameter increase of the subcutaneous mass up to 4 cm. Thoracic radiography and ultrasonography revealed pleural effusion and a lytic process in the fourth left rib. Furthermore, ultrasound examination revealed a mixed echogenic mobile structure with a diameter of around 2 cm floating within the pleural fluid of the left hemithorax close to the pericardium. The dog underwent surgery for an en bloc resection of the subcutaneous mass together with the fourth rib and the parietal pleura. Moreover, the left altered lung lobe, corresponding to the mobile structure detected by ultrasound, was removed. Based on cytological, histopathological, and immunohistochemical examinations, an invasive epithelioid pleural malignant mesothelioma was diagnosed

Galaxy Counts at 24 Microns in the SWIRE Fields

This paper presents galaxy source counts at 24 microns in the six Spitzer Wide-field InfraRed Extragalactic (SWIRE) fields. The source counts are compared to counts in other fields, and to model predictions that have been updated since the launch of Spitzer. This analysis confirms a very steep rise in the Euclidean-normalized differential number counts between 2 mJy and 0.3 mJy. Variations in the counts between fields show the effects of sample variance in the flux range 0.5-10 mJy, up to 100% larger than Poisson errors. Nonetheless, a "shoulder" in the normalized counts persists at around 3 mJy. The peak of the normalized counts at 0.3 mJy is higher and narrower than most models predict. In the ELAIS N1 field, the 24 micron data are combined with Spitzer-IRAC data and five-band optical imaging, and these bandmerged data are fit with photometric redshift templates. Above 1 mJy the counts are dominated by galaxies at z less than 0.3. By 300 microJy, about 25% are between z ~ 0.3-0.8, and a significant fraction are at z ~ 1.3-2. At low redshifts the counts are dominated by spirals, and starbursts rise in number density to outnumber the spirals' contribution to the counts below 1 mJy.Comment: 10 pages, 8 figures, accepted 3 November 2007 for publication in The Astronomical Journal, formatted with emulateapj styl

Complete Multiwavelength Characterization of Faint Chandra X-ray Sources Seen in the Spitzer Wide-Area IR Extragalactic (SWIRE) Survey

We exploit deep combined observations with Spitzer and Chandra of the SWIRE survey in the ELAIS-N1 region, to investigate the nature of the faint X-ray and IR sources in common, to identify AGN/starburst diagnostics, and to study the sources of the X-ray and IR cosmic backgrounds. In the 17'x17' area of the Chandra ACIS-I image there are 3400 SWIRE near-IR sources with 4 sigma detections in at least 2 IRAC bands and 988 sources detected at 24micron with MIPS brighter than 0.1 mJy. Of these, 102 IRAC and 59 MIPS sources have Chandra counterparts, out of a total of 122 X-ray sources present in the area with S(0.5-8 kev)>10^(-15) erg/cm^2/s. We have constructed SEDs for each source using data from the 4 IRAC wavebands, Chandra fluxes, and optical follow-up data in the wavebands U, g', r', i', Z, and H. We fit a number of spectral templates to the SEDs at optical and infrared wavelengths to determine photometric redshifts and spectral categories, and also make use of diagnostics based on the X-ray luminosities, hardness ratios, X-ray to infrared spectral slopes and optical morphologies. Although we have spectroscopic redshifts for only a minority of the Chandra sources, the available SEDs constrain the redshifts for most of the sample sources, which turn out to be typically at 0.5<z<2. We find that 39% of the Chandra sources are dominated by type-1 AGN emission, 23% display optical/IR spectra typical of type-2 AGNs, while the remaining 38% fraction show starburst-like or even normal galaxy spectra. Since we prove that all these galaxies are dominated by AGN emission in X-rays this brings the fraction of type-1 AGNs to be 80% of the type-2: even assuming that all the Chandra sources undetected by Spitzer are type-2 AGNs, the type-1 fraction would exceed 1/3 of the total population (abridged).Comment: Accepted for publication in AJ, March 2005 issu

SWIRE: The SIRTF Wide‐Area Infrared Extragalactic Survey

The SIRTF Wide-Area Infrared Extragalactic Survey (SWIRE), the largest SIRTF Legacy program, is a wide-area imaging survey to trace the evolution of dusty, star-forming galaxies, evolved stellar populations, and active galactic nuclei (AGNs) as a function of environment, from redshifts to the current z ∼ 3 epoch. SWIRE will survey seven high-latitude fields, totaling 60–65 deg2 in all seven SIRTF bands: Infrared Array Camera (IRAC) 3.6, 4.5, 5.6, and 8 mm and Multiband Imaging Photometer for SIRTF (MIPS) 24, 70, and 160 mm. Extensive modeling suggests that the Legacy Extragalactic Catalog may contain in excess of 2 million IR-selected galaxies, dominated by (1) ∼150,000 luminous infrared galaxies (LIRGs; LFIR 1 1011 L,) detected by MIPS (and significantly more detected by IRAC), ∼7000 of these with ; (2) 1 million IRAC- z 1 2 detected early-type galaxies (∼ with and ∼10,000 with ); and (3) ∼20,000 classical AGNs 5 2 # 10 z 1 1 z 1 2 detected with MIPS, plus significantly more dust-obscured quasi-stellar objects/AGNs among the LIRGs. SWIRE will provide an unprecedented view of the evolution of galaxies, structure, and AGNs. The key scientific goals of SWIRE are (1) to determine the evolution of actively star forming and passively evolving galaxies in order to understand the history of galaxy formation in the context of cosmic structure formation; (2) to determine the evolution of the spatial distribution and clustering of evolved galaxies, starbursts, and AGNs in the key redshift range over which much of cosmic evolution has occurred; and (3) to 0.5 ! z ! 3 determine the evolutionary relationship between “normal galaxies” and AGNs and the contribution of AGN accretion energy versus stellar nucleosynthesis to the cosmic backgrounds. The large area of SWIRE is important to establish statistically significant population samples over enough volume cells that we can resolve the star formation history as a function of epoch and environment, i.e., in the context of structure formation. The large volume is also optimized for finding rare objects. The SWIRE fields are likely to become the next generation of large “cosmic windows” into the extragalactic sky. They have been uniquely selected to minimize Galactic cirrus emission over large scales. The Galaxy Evolution Explorer will observe them as part of its deep 100 deg2 survey, as will Herschel. SWIRE includes ∼9 deg2 of the unique large-area XMM Large Scale Structure hard X-ray imaging survey and is partly covered by the UKIDSS deep J and K survey. An extensive optical/near-IR imaging program is underway from the ground. The SWIRE data are nonproprietary; catalogs and images will be released twice yearly, beginning about 11 months after SIRTF launch. Details of the data products and release schedule are presented

Type Ibn Supernovae Show Photometric Homogeneity and Spectral Diversity at Maximum Light

Type Ibn supernovae (SNe) are a small yet intriguing class of explosions whose spectra are characterized by low-velocity helium emission lines with little to no evidence for hydrogen. The prevailing theory has been that these are the core-collapse explosions of very massive stars embedded in helium-rich circumstellar material (CSM). We report optical observations of six new SNe Ibn: PTF11rfh, PTF12ldy, iPTF14aki, iPTF15ul, SN 2015G, and iPTF15akq. This brings the sample size of such objects in the literature to 22. We also report new data, including a near-infrared spectrum, on the Type Ibn SN 2015U. In order to characterize the class as a whole, we analyze the photometric and spectroscopic properties of the full Type Ibn sample. We find that, despite the expectation that CSM interaction would generate a heterogeneous set of light curves, as seen in SNe IIn, most Type Ibn light curves are quite similar in shape, declining at rates around 0.1 mag day-1 during the first month after maximum light, with a few significant exceptions. Early spectra of SNe Ibn come in at least two varieties, one that shows narrow P Cygni lines and another dominated by broader emission lines, both around maximum light, which may be an indication of differences in the state of the progenitor system at the time of explosion. Alternatively, the spectral diversity could arise from viewing-angle effects or merely from a lack of early spectroscopic coverage. Together, the relative light curve homogeneity and narrow spectral features suggest that the CSM consists of a spatially confined shell of helium surrounded by a less dense extended wind