Protocol for affinity purification-mass spectrometry interactome profiling in larvae of Drosophila melanogaster.

Many techniques exist for the identification of protein interaction networks. We present a protocol that relies on an affinity purification-mass spectrometry (AP-MS) approach to detect proteins that co-purify with a tagged bait of interest from Drosophila melanogaster larval muscles using the GAL4/upstream activating sequence (UAS) expression system. We also describe steps for the isolation and identification of protein complexes, followed by streamlined bioinformatics analysis for rapid and reproducible results.

Secretome Analysis of LMBC 162 Cultivated with Seeds Reveals CAZymes for Degradation of Lignocellulosic Biomass.

The analysis of the secretome allows us to identify the proteins, especially carbohydrate-active enzymes (CAZymes), secreted by different microorganisms cultivated under different conditions. The CAZymes are divided into five classes containing different protein families. is a thermophilic ascomycete, a source of many glycoside hydrolases and oxidative enzymes that aid in the breakdown of lignocellulosic materials.

Mycobacterium abscessus extracellular vesicles increase mycobacterial resistance to clarithromycin in vitro.

Nontuberculous Mycobacteria (NTM) are a group of emerging bacterial pathogens that have been identified in cystic fibrosis (CF) patients with microbial lung infections. The treatment of NTM infection in CF patients is challenging due to the natural resistance of NTM species to many antibiotics. Mycobacterium abscessus is one of the most common NTM species found in the airways of CF patients.

Intracellular iron accumulation facilitates mycobacterial infection in old mouse macrophages.

Aging has a significant impact on the immune system, leading to a gradual decline in immune function and changes in the body's ability to respond to bacterial infections. Non-tuberculous mycobacteria (NTM), also known as atypical mycobacteria or environmental mycobacteria, are commonly found in soil, water, and various environmental sources. While many NTM species are considered opportunistic pathogens, some can cause significant infections, particularly in individuals with compromised immune systems, such as older individuals.

Role of the Mycobacterium tuberculosis ESX-4 Secretion System in Heme Iron Utilization and Pore Formation by PPE Proteins.

Mycobacterium tuberculosis () is transmitted through aerosols and primarily colonizes within the lung. The World Health Organization estimates that kills ~1.4 million people every year.

Integrative proteomics and metabolomics profiling to understand the biochemical basis of beef muscle darkening at a slightly elevated pH.

Previous studies investigated the biochemical basis of dark-cutting conditions at elevated muscle pH (above 6), but the molecular basis at slightly above normal pH (between 5.6 and 5.8) is still unclear.

Dark-cutting beef mitochondrial proteomic signatures reveal increased biogenesis proteins and bioenergetics capabilities.

Mitochondria remain active in postmortem muscles and can influence meat color via oxygen consumption. Previous studies have shown that dark-cutting compared with normal-pH beef has greater mitochondrial protein and DNA content per gram of muscle tissue. However, the mechanism regulating mitochondrial content in dark-cutting vs.

'Walking in the shoes of our patients': a scoping review of healthcare professionals learning from the simulation of patient illness experiences.

Background: Health professionals who have experienced ill-health appear to demonstrate greater empathy towards their patients. Simulation can afford learners opportunities to experience aspects of illness, but to date, there has been no overarching review of the extent of this practice or the impact on empathic skills.

Objective: To determine from the evidence-what is known about simulation-based learning methods of creating illness experiences for health professions and the impact on their empathic skills.

Hyperglycemia-induced effects on glycocalyx components in the retina.

Purpose: Diabetic retinopathy is a vision-threatening complication of diabetes characterized by endothelial injury and vascular dysfunction. The loss of the endothelial glycocalyx, a dynamic layer lining all endothelial cells, contributes to several microvascular pathologies, including an increase in vascular permeability, leukocyte plugging, and capillary occlusion, and may drive the progression of retinopathy. Previously, a significant decrease in glycocalyx thickness has been observed in diabetic retinas.

Changes in composition and levels of hemolymph proteins during metamorphosis of Manduca sexta.

The tobacco hornworm, Manduca sexta, is a lepidopteran model species widely used to study insect biochemical processes. Some of its larval hemolymph proteins are well studied, and a detailed proteomic analysis of larval plasma proteins became available in 2016, revealing features such as correlation with transcriptome data, formation of immune complexes, and constitution of an immune signaling system in hemolymph. It is unclear how the composition of these proteins may change in other developmental stages.

Changes in glycolytic and mitochondrial protein profiles regulates postmortem muscle acidification and oxygen consumption in dark-cutting beef.

Dark-cutting beef is a condition in which beef fails to have a characteristic bright-red color when the cut surface is exposed to oxygen. However, the mechanistic basis for this occurrence is not clear. Protein expression profiles were compared between dark-cutting and normal-pH beef using LC-MS/MS-based proteomics.

'Sorry doctor….I didn't hear that….': phenomenological analysis of medical students' experiences of simulated hearing impairment through virtual reality.

Introduction: Hearing impairment is a common condition that can have a significant impact on an individual. Ineffective communication between such individuals and doctors remains an important barrier. There is a need to provide medical students with a deeper understanding of such challenges.

Proteomic and Metabolomic Analyses of a Tea-Tree Oil-Selected Small Colony Variant.

Tea tree oil (TTO) is hypothesized to kill bacteria by indiscriminately denaturing membrane and protein structures. A small colony variant (SCV) selected with TTO (SH1000-TTORS-1) demonstrated slowed growth, reduced susceptibility to TTO, a diminutive cell size, and a thinned cell wall. Utilizing a proteomics and metabolomics approach, we have now revealed that the TTO-selected SCV mutant demonstrated defective fatty acid synthesis, an alteration in the expression of genes and metabolites associated with central metabolism, the induction of a general stress response, and a reduction of proteins critical for active growth and translation.

Proteomic Profiling of Hsp90 Inhibitors.

Mass spectrometry assays demonstrate that Hsp90 inhibitors alter the expression of approximately one-quarter of the assayable proteome in mammalian cells. These changes are extraordinarily robust and reproducible, making "proteomics profiling" the gold standard for validating the effects of new Hsp90 inhibitors on cultured cells. Proteomics assays can also suggest novel hypotheses regarding drug mechanisms.

Hemolymph proteins of Anopheles gambiae larvae infected by Escherichia coli.

Anopheles gambiae is a major vector of human malaria and its immune system in part determines the fate of ingested parasites. Proteins, hemocytes and fat body in hemolymph are critical components of this system, mediating both humoral and cellular defenses. Here we assessed differences in the hemolymph proteomes of water- and E.

Myceliophthora thermophila M77 utilizes hydrolytic and oxidative mechanisms to deconstruct biomass.

Biomass is abundant, renewable and useful for biofuel production as well as chemical priming for plastics and composites. Deconstruction of biomass by enzymes is perceived as recalcitrant while an inclusive breakdown mechanism remains to be discovered. Fungi such as Myceliophthora thermophila M77 appear to decompose natural biomass sources quite well.

Changes in the Plasma Proteome of Manduca sexta Larvae in Relation to the Transcriptome Variations after an Immune Challenge: Evidence for High Molecular Weight Immune Complex Formation.

Manduca sextais a lepidopteran model widely used to study insect physiological processes, including innate immunity. In this study, we explored the proteomes of cell-free hemolymph from larvae injected with a sterile buffer (C for control) or a mixture of bacteria (I for induced). Of the 654 proteins identified, 70 showed 1.

Escherichia coli persister cells suppress translation by selectively disassembling and degrading their ribosomes.

Bacterial persisters are rare, phenotypically distinct cells that survive exposure to multiple antibiotics. Previous studies indicated that formation and maintenance of the persister phenotype are regulated by suppressing translation. To examine the mechanism of this translational suppression, we developed novel methodology to rapidly purify ribosome complexes from persister cells.

The anticancer drug AUY922 generates a proteomics fingerprint that is highly conserved among structurally diverse Hsp90 inhibitors.

AUY922 is a potent synthetic Hsp90 antagonist that is moving steadily through clinical trials against a small range of cancers. To identify protein markers that might measure the drug's effects, and to gain understanding of mechanisms by which AUY922 might inhibit the proliferation of leukemia cells, we characterized AUY922's impacts on the proteomes of cultured Jurkat cells. We describe a robust and readily assayed proteomics fingerprint that AUY922 shares with the flagship Hsp90 inhibitors 17-DMAG and radicicol.

Allied health students' perceptions of effective clinical instruction.

Students of allied health who have positive clinical education experiences often accept employment offered by their clinical sites. Most allied health professionals are not trained to be educators; to improve student clinical experiences, students were surveyed regarding their learning experiences during clinical internships. This study examined the teaching abilities and professional development training needs of clinical supervisors as identified by student perceptions.

What future health care providers will need to know about child abuse and neglect.

Child abuse and neglect have reached epidemic proportions. State and federal laws mandate that allied health professionals report suspected child abuse and neglect. Academic and continuing education programs could assist allied health professionals to be legally responsible, mandated reporters of child abuse and neglect.

Training health care providers to be educators.

Background: Students in the allied health field who have a positive clinical education experience often accept employment opportunities offered by their clinical site. Most allied health professionals are not trained to be educators. Professional development in the area of teaching could assist the professional to be a more effective teacher and therefore improve the clinical experience.

Novel subunits of the mammalian Hsp90 signal transduction chaperone.

As one of the major cellular chaperones, Hsp90 plays diverse roles in supporting and regulating wild-type and oncogenic signal transduction proteins. Hsp90 function itself is regulated by its various nonsubstrate subunits. To define Hsp90's predominant in vivo functions and the mechanisms for regulating this function, the human Hsp90 interactome was characterized using gel-based proteomics techniques.

Does requiring continuing education units for professional licensing renewal assure quality patient care?

Although most states in the United States require health care professionals to complete continuing education units (CEUs) for licensure renewal, little evidence to date has established a relationship between completing CEUs and clinical competency. Considering the high cost of health care delivery and services, it would be prudent for both managers and consumers of health care to review the costs and benefits of requiring CEUs for professional licensing renewal. This study features an extensive review of the literature to analyze the supportive as well as the opposing views of mandatory CEUs for professional license renewal.