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Among several potential animal models that can be used for adipogenic studies, Wagyu cattle is the one that presents unique molecular mechanisms underlying the deposit of substantial amounts of intramuscular fat. As such, this review is focused on current knowledge of such mechanisms related to adipose tissue deposition using Wagyu cattle as model. So abundant is the lipid accumulation in the skeletal muscles of these animals that in many cases, the muscle cross-sectional area appears more white (adipose tissue) than red (muscle fibers). This enhanced marbling accumulation is morphologically similar to that seen in numerous skeletal muscle dysfunctions, disease states and myopathies; this might indicate cross-similar mechanisms between such dysfunctions and fat deposition in Wagyu breed. Animal models can be used not only for a better understanding of fat deposition in livestock, but also as models to an increased comprehension on molecular mechanisms behind human conditions. This revision underlies some of the complex molecular processes of fat deposition in animals. [ABSTRACT FROM PUBLISHER]

A 45-d backgrounding study was conducted at the E.V. Smith Research Center in Shorter, AL to determine animal performance differences of pre-conditioned beef calves fed annual ryegrass (Lolium multiflorum) baleage, bermudagrass (Cynodon dactylon) hay, or corn (Zea mays) silage-based diets. Annual ryegrass (cv. Marshall) was harvested for baleage on 22 Apr 2015 at the late boot stage of maturity. The forage was allowed to wilt for 48 h until it achieved 60% moisture, baled and wrapped. Tifton 85 bermudagrass used for the study was harvested at a 4 to 5 wk interval in early summer 2015. Corn silage was harvested at the full dent stage of maturity on 15 Jul 2015, chopped, and stored until the initiation of the feeding trial. Forage concentrations of CP, ADF, and NDF (% DM) were 11.3, 35.6, and 57.2 for annual ryegrass baleage, 13.5, 35.8, and 63.9 for bermudagrass hay, and 5.8, 23.6, and 41.0 for corn silage, respectively. Based on forage quality, calves were supplemented with an energy-protein based ration (50:50 soybean hulls and corn gluten feed for baleage and hay treatments, and 85% corn, 15% cottonseed meal mix for corn silage treatments) to target 0.9 kg/day ADG according to NRC (2000) recommendations. The 45-d background trial began on 9 September 2015 after animals were sorted and acclimated to the diets. 108 weaned calves [heifers (n = 54; mean initial BW 283 kg) and steers (n = 54; mean initial BW 284 kg)] were placed into nine pens (n = 12/pen, 3 pens/treatment). Sex was distributed evenly across treatments. Animals were weighed on d 0, 22, and 44, and the study concluded on 23 October 2015. Animal performance measures were analyzed using PROC Mixed in SAS 9.4 as a completely randomized design, and pen was the experimental unit. Mean initial and final BW of the animals did not differ (P = 0.50 and P = 0.99, respectively) across treatments. Average daily gain for annual ryegrass baleage, bermudagrass hay, and corn silagebased diets were 0.61 kg/day, 0.72 kg/day, and 0.72 kg/day, respectively, and did not differ across treatments (P = 0.57). Based on these results, these forage options achieved a similar level of gain when supplemented for preconditioning beef calves. However, an economic analysis is needed to determine feasibility and potential break-even costs for using these options in beef operations. [ABSTRACT FROM AUTHOR]

A 45-d backgrounding study was conducted at the E.V. Smith Research Center in Shorter, AL to determine animal performance differences of pre-conditioned beef calves fed annual ryegrass (Lolium multiflorum) baleage, bermudagrass (Cynodon dactylon) hay, or corn (Zea mays) silage-based diets. Annual ryegrass (cv. Marshall) was harvested for baleage on 22 Apr 2015 at the late boot stage of maturity. The forage was allowed to wilt for 48 h until it achieved 60% moisture, baled and wrapped. Tifton 85 bermudagrass used for the study was harvested at a 4 to 5 wk interval in early summer 2015. Corn silage was harvested at the full dent stage of maturity on 15 Jul 2015, chopped, and stored until the initiation of the feeding trial. Forage concentrations of CP, ADF, and NDF (% DM) were 11.3, 35.6, and 57.2 for annual ryegrass baleage, 13.5, 35.8, and 63.9 for bermudagrass hay, and 5.8, 23.6, and 41.0 for corn silage, respectively. Based on forage quality, calves were supplemented with an energy-protein based ration (50:50 soybean hulls and corn gluten feed for baleage and hay treatments, and 85% corn, 15% cottonseed meal mix for corn silage treatments) to target 0.9 kg/day ADG according to NRC (2000) recommendations. The 45-d background trial began on 9 September 2015 after animals were sorted and acclimated to the diets. 108 weaned calves [heifers (n = 54; mean initial BW 283 kg) and steers (n = 54; mean initial BW 284 kg)] were placed into nine pens (n = 12/pen, 3 pens/treatment). Sex was distributed evenly across treatments. Animals were weighed on d 0, 22, and 44, and the study concluded on 23 October 2015. Animal performance measures were analyzed using PROC Mixed in SAS 9.4 as a completely randomized design, and pen was the experimental unit. Mean initial and final BW of the animals did not differ (P = 0.50 and P = 0.99, respectively) across treatments. Average daily gain for annual ryegrass baleage, bermudagrass hay, and corn silagebased diets were 0.61 kg/day, 0.72 kg/day, and 0.72 kg/day, respectively, and did not differ across treatments (P = 0.57). Based on these results, these forage options achieved a similar level of gain when supplemented for preconditioning beef calves. However, an economic analysis is needed to determine feasibility and potential break-even costs for using these options in beef operations. [ABSTRACT FROM AUTHOR]

Comparative, dose-dependent analysis of interactions between small molecule drugs and their targets, as well as off-target interactions, in complex proteomes is crucial for selecting optimal drug candidates. The affinity of small molecules for targeted proteins is largely dictated by interactions between amino acid side chains and these drugs. Thus, studying drug-protein interactions at an amino acid resolution provides a comprehensive understanding of the drug selectivity and efficacy. In this study, we further refined the site-specific activity-based protein profiling strategy (ABPP), PhosID-ABPP, on a timsTOF HT mass spectrometer. This refinement enables dual dose-dependent competition of inhibitors within a single cellular proteome. Here, a comparative analysis of two activity-based probes (ABPs), developed to selectively target the epidermal growth factor receptor (EGFR), namely, PF-06672131 (PF131) and PF-6422899 (PF899), facilitated the simultaneous identification of ABP-specific binding sites at a proteome-wide scale within a cellular proteome. Dose-dependent probe-binding preferences for proteinaceous cysteines, even at low nanomolar ABP concentrations, could be revealed. Notably, in addition to the intrinsic affinity of the electrophilic probes for specific sites in targeted proteins, the observed labeling intensity is influenced by several other factors. These include the efficiency of cellular uptake, the stability of the probes, and their intracellular distribution. While both ABPs showed comparable labeling efficiency for EGFR, PF131 had a broader off-target reactivity profile. In contrast, PF899 exhibited a higher labeling efficiency for the ERBB2 receptor and bound to catalytic cysteines in several other enzymes, which is likely to disrupt their catalytic activity. Notably, PF131 effectively labeled ADP/ATP translocase proteins at a concentration of just 1 nm, and we found this affected ATP transport. Analysis of the effect of PF131 and its parent inhibitor Afatinib on murine translocase SLC25A4 (ANT1)-mediated ATP transport strongly indicated that PF131 (10 μM) partially blocked ATP transport. Afatinib was less efficient at inhibiting ATP transport by SLC25A4 than PF131, and the reduction of ATP transport by Afatinib was not significant. Follow-up analysis is required to evaluate the affinity of these inhibitors for ADP/ATP translocase SLC25A4 in more detail. Additionally, the analysis of different binding sites within the EGF receptor and the voltage-dependent anion channel 2 revealed secondary binding sites of both probes and provided insights into the binding poses of inhibitors on these proteins. Insights from the PhosID-ABPP analysis of these two ABPs serve as a valuable resource for understanding drug on- and off-target engagement in a dose- and site-specific manner.

Most drug molecules target proteins. Identification of the exact drug binding sites on these proteins is essential to understand and predict how drugs affect protein structure and function. To address this challenge, we developed a strategy that uses immobilized metal-affinity chromatography-enrichable phosphonate affinity tags, for efficient and selective enrichment of peptides bound to an activity-based probe, enabling the identification of the exact drug binding site. As a proof of concept, using this approach, termed PhosID-ABPP (activity-based protein profiling), over 500 unique binding sites were reproducibly identified of an alkynylated afatinib derivative (PF-06672131). As PhosID-ABPP is compatible with intact cell inhibitor treatment, we investigated the quantitative differences in approachable binding sites in intact cells and in lysates of the same cell line and observed and quantified substantial differences. Moreover, an alternative protease digestion approach was used to capture the previously reported binding site on the epidermal growth factor receptor, which turned out to remain elusive when using solely trypsin as protease. Overall, we find that PhosID-ABPP is highly complementary to biotin-based enrichment strategies in ABPP studies, with PhosID-ABPP providing the advantage of direct activity-based probe interaction site identification., Competing Interests: Conflict of interest The authors declare no competing interest., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Tight regulation of protein translation drives the proteome to undergo changes under influence of extracellular or intracellular signals. Despite mass spectrometry-based proteomics being an excellent method to study differences in protein abundance in complex proteomes, analyzing minute or rapid changes in protein synthesis and abundance remains challenging. Therefore, several dedicated techniques to directly detect and quantify newly synthesized proteins have been developed, notably puromycin-based, bio-orthogonal noncanonical amino acid tagging-based, and stable isotope labeling by amino acids in cell culture-based methods, combined with mass spectrometry. These techniques have enabled the investigation of perturbations, stress, or stimuli on protein synthesis. Improvements of these methods are still necessary to overcome various remaining limitations. Recent improvements include enhanced enrichment approaches and combinations with various stable isotope labeling techniques, which allow for more accurate analysis and comparison between conditions on shorter timeframes and in more challenging systems. Here, we aim to review the current state in this field., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Delirium is a serious neuropsychiatric syndrome, which requires timely treatment. However, it is easily missed, especially in older patients with premorbid cognitive disorders., Objectives: The aim of this study is to investigate the prevalence and risk factors of delirium in older outpatients with and without dementia., Method: We assessed 444 patients referred to the memory clinic of a psychiatric hospital between March 2013 and March 2014. Demographic information, medical history, impairments in daily living activities and referral information were registered. Patients underwent a psychiatric examination using the Delirium Rating Scale-Revised-98 and cognitive tests, a physical examination and laboratory tests. We recorded medication use and changes before and after the onset of symptoms., Results: Among the 444 outpatients, 85 had probable delirium (prevalence of 19%), and 10 had subsyndromal delirium (2%). The most common triggers were infection (42%), drug-intoxication or withdrawal (22%), and metabolic/endocrine disturbance (12%). Age (OR 1.07, 95% CI 1.02-1.11) and prior delirium (OR 3.34, 95% CI 1.28-8.69) were independent non-modifiable factors associated with an increased risk of delirium. The only independent modifiable risk factor was infection (OR 17.31, 95% CI 8.44-35.49)., Conclusions: A delirium was detected in one of five patients referred for dementia screening. Most patients could be treated at home. Age and prior delirium were predictive of an increased risk of delirium., (© 2020 The Authors. International Journal of Geriatric Psychiatry published by John Wiley & Sons Ltd.)

Chlamydial-induced cystitis in the koala (Phascolarctos cinereus) is currently treated by antibiotics. However, while reducing the chlamydial load, this treatment can also lead to gastrointestinal complications and death. Development of alternative treatments, such as a therapeutic chlamydial vaccine, are hindered by the lack of detailed understanding of the innate immune response to chlamydial clearance and disease regression during antibiotic treatment. Through clinical, microbiological and transcriptomic approaches, disease regression, bacterial clearance and innate immune responses were mapped in koalas with signs of chlamydial-induced cystitis while receiving anti-chlamydial antibiotics. Significant reduction in the signs of cystitis were observed during and post antibiotic treatment. This was observed as a thinning of the bladder wall and complete reversal of urinary incontinence. Transcriptomic analysis before treatment, at the end of treatment and prior to release identified significant down-regulation of specific genes involved in 21 biological pathways. Of these, the chemokine receptor signalling and NOD-like receptor signalling pathways where identified as important markers of inflammation. Specific genes within these pathways (NCF1 and NOX2) were significantly down-regulated, suggesting a decrease in reactive oxygen species production. Through the monitoring of specific clinical and transcriptomic markers, these findings allow detailed profiling of the clinical response to therapeutic vaccination in koalas with current signs of disease. This also adds to our understanding of innate immune responses to chlamydial infections and indicates that chlamydial-induced cystitis in the koala is linked to the regulation of reactive oxygen pathways., Competing Interests: The authors have declared that no competing interests exist.

Tuberculosis (TB) remains a major global public health problem that has become a crisis fuelled by HIV and the increasing occurrence of antimicrobial resistance. What has been termed the biosocial nature of TB challenges effective control of the disease. Yet, biosocial interactions involved in the persistence of TB in diverse settings are difficult to systematically account for. The recently developed framework of syndemics provides a way to capture how complex health problems result from the interactions between diseases such as HIV and TB, and harmful social conditions such as unemployment, malnutrition and substance abuse. This article advances the syndemics scholarship by examining health conditions that cluster together with TB in the Russian Federation, by eliciting a set of social processes that precipitate this clustering and exacerbate health outcomes, and by analysing interactions between these health conditions and social processes. To provide an account of this complexity, the article takes a qualitative approach and draws on the perspectives and experiences of people with TB. The results demonstrate emergence of a syndemic of stress, substance abuse, TB and HIV that is sustained by poverty, occupational insecurity, marginalization and isolation. Frictions between the narrow focus of the health care system on TB and the wider syndemic processes in which the lives of many persons with TB are embedded, contribute to poorer health outcomes and increase the risks of developing drug resistance. Finally, the article argues that the large-scale and impersonal forces become embodied as individual pathology through the crucial interface of the ways in which persons experience and make sense of these forces and pathologies. Qualitative research is needed for the adequate analysis of this biosocial complexity in order to provide a solid basis for responses to TB-centred syndemics in various settings.