Shellfish allergy is a major global health concern, with prawns representing the most common trigger. Black tiger prawn (Penaeus monodon, BTP) and white leg prawn (Litopenaeus vannamei, WLP) dominate global consumption. Here, label-free shotgun proteomics combined with in-silico allergenicity prediction was applied to identify and compare the allergen profiles of BTP and WLP. Raw prawn extracts were analyzed by LC-MS/MS, and allergens were identified and predicted using AllerCatPro. The relative abundance of allergens was assessed using iBAQ%, and the differential abundance was evaluated using LFQ intensity. Although allergens accounted for only ∼4% of identified proteins, they represented 34-38% of total protein abundance, highlighting their immunological relevance. Myosin light chain was the most abundant allergen, followed by arginine kinase, sarcoplasmic calcium-binding protein, and tropomyosin in both species. Four low-abundance proteins were identified as novel allergen candidates. Notable differences in allergen profiles between the two species were observed at the level of allergen orthologs, isoforms, and variants, with nine uniquely identified in BTP and seven in WLP. Together, these findings reveal species-specific variations with implications for improved diagnostic strategies, therapeutic approaches, and allergen detection methods for shellfish allergy.

Allergic reactions to fish pose complex food safety challenges, driven by species diversity and underexplored intraspecies variability. This study comparatively examines allergen profiles in Malabar red snappers (Lutjanus malabaricus, n = 39) across body sizes, anatomical regions and production origins using SDS-PAGE, immunoblotting and quantitative mass spectrometry. Protein profiles varied greatly by fish size and muscle region, but not by origin. Smaller fish contained higher levels of major allergen parvalbumin and creatine kinase, while larger fish exhibited elevated levels of heat-labile allergens enolase, aldolase, and glyceraldehyde-3-phosphate dehydrogenase. Parvalbumin levels were highest in head, followed by belly, dorsal, and tail. Greatest variation was observed for the three heat-stable allergens - parvalbumin, tropomyosin, and collagen. Minimal origin-dependent differences affected 2 of 11 registered fish allergens. We established an integrated proteomics workflow for systematic allergenicity assessments to uncover intraspecies variability and provide foundational knowledge for understanding intraspecies variability to improve food safety strategies.

Type 2 diabetes and obesity are commonly accompanied by metabolic dysfunction-associated steatotic liver disease (MASLD), increasing the risk of developing metabolic dysfunction-associated steatohepatitis (MASH) and fibrosis. The early stages of MASLD are characterized by dysfunctional lipid metabolism, including remodelling of the hepatic lipidome. In this context, reductions in hepatic phosphatidylserine (PS) have been associated with increased hepatic steatosis, inflammation and fibrosis. In this study, we investigated the impact of dietary PS supplementation on liver function and systemic metabolic homeostasis in mice with hepatic steatosis and MASH. Taking advantage of the MUP-uPA mouse model, including wild-type (WT) mice with hepatic steatosis and MUP-uPA mice with MASH and fibrosis, we show that PS supplementation reduces hepatic triglyceride accumulation, inflammation and fibrosis in male MUP-uPA mice. Supporting these data, PS supplementation suppresses fibrogenic gene expression in LX-2 hepatic stellate cells. We further show that PS supplementation improves glycaemic control and insulin sensitivity in male and female mice, which is associated with enhanced insulin signalling in muscle and liver, despite a pronounced suppression of glycolysis, glucose oxidation and glycogen breakdown in liver, muscle and/or adipose tissue. Metabolic flux analysis suggests a shift in substrate utilization, favouring fatty acid metabolism, particularly in muscle, while further pointing to marked improvements in mitochondrial function and oxidative capacity. These findings indicate that PS exerts multifaceted benefits by improving both MASH and whole-body glucose homeostasis, independent of conventional oxidative glucose metabolism. Our results support further investigation into dietary PS as a potential complementary strategy for MASH and glycaemic control.

The ubiquitin-specific protease (USP) USP9X is a human deubiquitinase (DUB) with a large number of described targets and cellular roles. In cancer, USP9X is found as an oncogene or as a tumour suppressor depending on context, and its utility as a target for cancer therapy remains unclear. We here describe WEHI-092, a piperazine-based USP9X-specific small-molecule inhibitor, which binds to a unique region in the USP9X Fingers-subdomain, distinct from known DUB-inhibitor binding sites. Using proteomics and ubiquitinomics, we show that USP9X targets distinct substrates compared to USP7, yet the substrate profile of USP9X varies significantly across cancer cell lines. We reveal a core set of 17 proteins commonly regulated by USP9X in most cell lines, which we consider as proximal biomarkers for USP9X inhibition. Consistent with proteomics, we show in unrelated cell lines that WEHI-092 treatment arrests the cell cycle in metaphase without inducing cell death. This explains growth suppression in long-term clonogenic assays in most cancer cell lines, and positions USP9X inhibitors as a new class of selective mitotic poisons.

Preeclampsia, a severe pregnancy-induced disorder unique to humans, affects ∼2%-8% of pregnancies globally. Strong evidence supports placental dysfunction as central to preeclampsia; however, there is inadequate understanding of the precise pathogenesis of preeclampsia. In this study, we present a comprehensive multi-omics analysis of early pregnancy placental biopsies (chorionic villus samples) from pregnancies that later developed preterm/term preeclampsia, compared to normotensive controls. Using an integrative multivariate approach, we uncovered distinct molecular signatures associated with preeclampsia. Preterm preeclampsia was strongly associated with dysregulated lipoprotein metabolism, while term preeclampsia exhibited alterations in inflammatory pathways, Notch/Kit signaling, and ribosome assembly. These results challenge the prevailing notion that term preeclampsia is unrelated to early placental pregnancy dysfunction. To validate our findings, we focused on melanophilin, a gene downregulated in the early pregnancy placenta of term preeclampsia. Melanophilin expression was reduced during cytotrophoblast syncytialization; however, excessive loss disrupted syncytiotrophoblast function, triggering the production of factors known to drive preeclampsia. Our study provides critical insights into the early pregnancy aberrations underlying preterm and term preeclampsia, paving the way for the development of predictive biomarkers and targeted preventative treatments. This work represents a significant step toward unraveling the complex etiology of preeclampsia and improving maternal and perinatal health outcomes.

Seafood allergy is complex due to extensive species diversity, posing major challenges in food safety assessments, clinical diagnosis and dietary management. However, the absence of established workflows to resolve allergenomes limits correlations between allergen abundance, clinical sensitisation, and consumer risk. Mass spectrometry (MS)-based proteomics overcomes limitations of conventional immunoassay allergen detection by enabling unbiased protein identification and quantification, including allergen isoforms and low-abundance proteins within complex matrices. An integrated workflow combining immunological analyses, liquid chromatography-MS/MS proteomics, and bioinformatics was developed to characterise allergenomes across eight commonly consumed Asia-Pacific fish species. Comprehensive allergen profiles were established using in silico allergenicity predictions with AllerCatPro, combined with immunological validations using allergen-specific antibody and pooled patient sera. Across eight species, 529-1012 protein groups were identified, including all 11 fish muscle allergens, with pronounced interspecies differences in allergen composition and isoform distribution. The major fish pan-allergen parvalbumin was the most abundant allergen of most species and varied in abundance by up to 7-fold. Mackerel displayed a distinct low-parvalbumin profile with enriched metabolic allergens. Tissue heating induced a consistent shift toward enrichment of heat-stable and tissue-retained allergens, particularly parvalbumin, tropomyosin, and collagen. In contrast, heating of raw extracts generated more variable and species-specific retention of selected proteins, including heat-labile metabolic enzymes. In silico analysis predicted 14 proteins with strong allergenicity evidence for further validation. Fish allergenomes are species-specific and processing-dependent, positioning quantitative proteomics as a powerful platform for improved molecular risk assessment, and the development of representative diagnostic and food safety reference materials.

Polyamines are small, polycationic molecules with amino groups that are present in most living organisms. Studies indicate that polyamines increase general protein synthesis and are essential for efficient translation. While progress has been made in understanding the role of polyamines in translation in bacteria and mammals, their contribution and mode of action in plants remain largely unexplored. In a previous study, we found that putrescine (Put) and the pathogen-associated molecular pattern (PAMP) from bacterial flagellin (flg22) transcriptionally induced ribosome biogenesis in plants. Here we examined the impact of polyamines (Put and spermine, Spm) and flg22 on ribosome complex formation in Arabidopsis. Our results indicate that polyamines, flg22 and their combinations increase the abundance of actively translating polysomes. Riboproteomic analyses revealed that polyamines and flg22 trigger differential changes in the accumulation of ribosomal proteins, which are structurally confined in response to Put. Importantly, Put was found binding to non-translating and actively translating ribosomes, suggesting that this polyamine has a role in functional aspects of translation, such as stabilization and/or remodeling of polysomal complexes. Additional global proteomics analyses in polyamine biosynthesis mutants revealed that lower Put availability triggers changes in proteins associated with ribonucleoprotein complex binding and biogenesis. Overall, our findings highlight the effect of polyamines and flg22 on shaping the ribosomal protein composition of actively translating ribosomes in plants.

The rabies virus (RABV) phosphoprotein (P protein) has multiple functions, including acting as the essential non-catalytic cofactor of the viral polymerase (L protein) for genome replication and transcription; the principal viral antagonist of the interferon (IFN)-mediated innate immune response; and the chaperone for the viral nucleoprotein (N protein). Although P protein is known to undergo phosphorylation by cellular kinases, the location and functions of the phosphorylation sites remains poorly defined. Here, we report the identification by mass-spectrometry (MS) of residues of P protein that are modified by phosphorylation in mammalian cells, including several novel sites. Analysis of P protein with phospho-mimetic and phospho-inhibitory mutations of three novel residues/clusters that were commonly identified by MS (Ser48, Ser183/187, Ser217/219/220) indicate that phosphorylation at each of these sites does not have a major influence on nuclear trafficking or antagonistic functions toward IFN signalling pathways. However, phosphorylation of Ser48 in the N-terminus of P protein impaired function in transcription/replication and in the formation of replication structures that contain complexes of P and N proteins, suggestive of altered interactions of these proteins. The crystal structure of P protein containing the S48E phospho-mimetic mutation indicates that Ser48 phosphorylation facilitates the binding of residues 41–52 of P protein into the RNA-binding groove of non-RNA-bound N protein (N0), primarily through the formation of a salt bridge with Arg434 of N protein. These data indicate that Ser48 modification regulates the cycling of P-N0 chaperone complexes that deliver N protein to RNA to enable transcription/replication, such that enhanced interaction due to S48E phospho-mimetic mutation reduces N protein delivery to the RNA, inhibiting subsequent transcription/replication processes. These data are, to our knowledge, the first to implicate phosphorylation of RABV P protein in conserved replication functions of the P gene.

The eukaryotic ribosome is highly modified by protein methylation, yet many of the responsible methyltransferases remain unknown. Here, we identify SET and MYND domain-containing protein 5 (SMYD5) as a ribosomal protein methyltransferase that catalyzes trimethylation of RPL40/eL40 at lysine 22. Through a systematic mass spectrometry-based approach, we identify 12 primary sites of protein methylation in ribosomes from K562 cells, including at RPL40 K22. Through in vitro methylation of synthetic RPL40 using fractionated lysate, we then identify SMYD5 as a candidate RPL40 K22 methyltransferase. We show that recombinant SMYD5 has robust activity toward RPL40 K22 in vitro and that active site mutations ablate this activity. Knockouts of SMYD5 in K562 cells show a complete loss of RPL40 K22 methylation and decreased polysome levels. We show that SMYD5 does not methylate histones in vitro, and by systematic analysis of its recognition motif, we find that SMYD5 requires a KXY motif for methylation, explaining its lack of activity toward histones.

BACKGROUND:Although stress is a known antecedent of psychiatric illness, the effect of stressors in animal models of obsessive-compulsive disorder (OCD) has not been comprehensively examined. METHOD:We investigated the effects of chronic early-life social isolation stress (SIS) in the SAPAP3 knockout (KO) mouse model of obsessive-compulsive (OC)-like behaviour. We assessed the behavioural responses to 6 weeks of social isolation stress (starting from 5 weeks of age), focusing on measures of anxiety-like and compulsive-like behaviours. We also determined proteomic expression profiles in the striatum and prefrontal cortex (PFC) in SAPAP3 KO mice and wild-type (WT) controls. RESULTS:As expected, we found that SAPAP3 KO mice exhibited an obsessive-compulsive-like phenotype, grooming excessively and displaying anxiety-like behaviour when compared to WT mice. Furthermore, we discovered that social isolation stress worsened compulsive grooming in male SAPAP3 KO mice, and exacerbated anxiety-like behaviour in both male and female SAPAP3 KO animals. Finally, we also revealed molecular changes closely associated with these behavioural responses to stress, with evidence of male-predominant stress-induced changes to the proteomic profile in the PFC of SAPAP3 KO animals (male-SIS WT vs KO comparison revealed 19 differentially expressed proteins, whereas the female-SIS WT vs KO comparison showed 5 differentially expressed proteins). Notably, we found minimal changes to the proteome in the striatum of SAPAP3 KO mice. CONCLUSION:Our findings show that SAPAP3 KO mice are susceptible to the deleterious effects of social isolation stress, and suggest that the PFC may be a particularly stress-sensitive node relevant to obsessive-compulsive-like behaviour.

BackgroundCurrent diagnostic methods have limitations in early prediction of dementia.ObjectiveDevelop an early screening tool to identify persons at high risk of dementia for early intervention.MethodsWe examined amyloid-β protein precursor (AβPP) and its fragments in urine from cognitively normal controls (CNs) and patients with mild cognitive impairment (MCI) or Alzheimer's disease (AD)-dementia using western blotting with different antibodies and developed a colloidal gold lateral flow-based qualitative strip.ResultsCompared with CNs, the amounts of various AβPP fragments with molecular weights of approximately 14, 28, 56, and 68 kDa in patients were greater. We therefore used the strip to detect urine Aβ-containing AβPP fragments and evaluated its potential in multiple aged cohorts from 11 cities in Jiangsu Province, China (n = 4418); Sichuan Provincial People's Hospital (Chengdu, China; n = 408); and the Australian Imaging, Biomarker and Lifestyle Study (AIBL; n = 367). Unitizing Aβ-binding phagocytosis-promoting peptides, the strips showed increasing positivity (9.5-16.9%) with ageing in the Jiangsu cohort and good clinical performance in the Chengdu cohort (κ = 0.704). Significant differences between CNs and patients were found in the AIBL cohort with negative Aβ-PET, those with the slope of Aβ-PET change <1 centiloid per year, those under 75 years of age, or those with a body mass index of 25-30.ConclusionsOur data indicate the high potential of urine AβPP fragments as biomarkers for MCI and dementia at an early stage and warrant further longitudinal studies.

Type 2 diabetes is a chronic metabolic disorder characterised by insulin resistance and sustained hyperglycaemia, and is a major cause of blindness, kidney failure, heart attacks and stroke. Our team has recently identified hexosaminidase A (HEXA) as an endocrine factor secreted by the liver that regulates sphingolipid metabolism in skeletal muscle. Specifically, HEXA converts GM2 to GM3 gangliosides within cell-surface lipid rafts. Remodelling of ganglioside composition by HEXA enhances IGF1 signalling in skeletal muscle, increasing muscle glucose uptake and improving blood glucose control. We produced a long-acting HEXA-FC fusion protein (murine HEXA and the fragment crystallisable [FC] region from IgG1) and evaluated the effects of chronic bi-weekly HEXA-FC administration (1 mg/kg body weight) on glycaemic control in C57BL/6 mice with diet-induced obesity and insulin resistance and the db/db mouse model of severe type 2 diabetes. Outcome measures included glucose and insulin tolerance, including a stable isotope-labelled GTT and assessment of tissue-specific glucose disposal, as well as proteomics analysis to define changes in skeletal muscle metabolism. Chronic administration of a long-acting recombinant HEXA-FC fusion protein led to improvements in random blood glucose, fasting blood glucose and glucose tolerance, driven by increased glucose disposal into skeletal muscle, effects that were associated with enhancement of IGF1 signalling in muscle. Given that skeletal muscle is a primary site of insulin resistance in individuals with type 2 diabetes, HEXA-FC protein therapy may open new avenues for therapeutic advancement in type 2 diabetes.

RNA viruses encode multifunctional proteins to overcome limited genomic capacity and mediate diverse processes in viral replication and host cell modulation. The rabies virus P gene encodes full-length P1 protein and the truncated isoform, P3, which acquires phenotypes absent from P1, including interactions with cellular membrane-less organelles (MLOs) formed by liquid-liquid phase separation (LLPS). This gain-of-function suggests that isoform multifunctionality arises not only from discrete functions of protein modules/domains, but also from conformational regulation involving interactions of the globular C-terminal domain and N-terminal intrinsically disordered regions (IDRs). The precise mechanisms underlying gain-of-function, however, remain unresolved. Here, we compare the structure and function of P1 and P3, identifying isoform-specific long-range intra-protomer interactions between the IDRs and C-terminal domain that correlate with conformational states, LLPS behavior, and subcellular localization. Mutations in P3 that alter MLO interactions correspondingly modulate these interactions. P1 and P3 can interact with similar/overlapping sets of MLO-associated proteins and have similar LLPS capacity, but only P3 binds RNA, and this interaction correlates with gain-/loss-of-function mutations. Our findings reveal that conformational differences in isoforms regulate LLPS behavior and contribute to protein-RNA interactions, which controls access to host LLPS structures, uncovering a previously unrecognized strategy in P protein multifunctionality.

Contractile myofibroblasts immersed in stiffened remodeled extracellular matrix characterize fibrotic lesions in idiopathic pulmonary fibrosis. Lipofibroblasts are lipid droplet-containing interstitial fibroblasts that support functional homeostasis of the developing and adult lung. We show that stiff substrates augment myofibroblast differentiation and extracellular matrix production in vitro under basal conditions and following TGF-β1 (transforming growth factor-β1) incubation when cultured on tissue culture plastic, whereas culture in soft microenvironments (as spheroids or on soft collagen-coated substrate) redirects myofibroblasts to a lipofibroblast-like phenotype (identified by expression of ADRP [adipose differentiation-related protein] and intracellular lipid droplets), with reduced basal α-SMA (α-smooth muscle actin), collagen I, vimentin, and fibronectin expression. The fibrogenic effects of TGF-β1 are prevented in fibroblasts cultured in soft settings. Global proteomics showed similar numbers of TGF-β1-induced differentially expressed proteins in stiff and soft settings (271 and 436, respectively). Of these, only 33 were similarly altered by TGF-β1; 200 were exclusively altered by TGF-β1 in the stiff setting and 365 in the soft setting; 38 showed opposite responses. Reductions in YAP/TAZ, β-catenin, and SMAD expression and their limited nuclear levels in soft settings may explain the "afibrogenic" characteristic of these lipofibroblasts. Thus, in spheroids of lipofibroblasts, TGF-β1 intracellular signaling is redirected and uncoupled from fibrogenesis, including YAP/TAZ, β-catenin, and SMAD. Understanding the proximal causal mechanotransduction signaling networks that are differentially active in soft and stiff microenvironments may reveal novel drug targets for fibrosis treatment.

Abstract Background Nippostrongylus brasiliensis—a nematode of rodents—is commonly used as a model to study the immunobiology of parasitic nematodes. It is a member of the Strongylida—a large order of socioeconomically important parasitic nematodes of animals. Lipids are known to play essential roles in nematode biology, influencing cellular membranes, energy storage and/or signalling. Methods The present investigation provides a comprehensive, untargeted lipidomic analysis of four developmental stages/sexes (i.e. egg, L3, adult female and adult male stages) of N. brasiliensis utilising liquid chromatography coupled to mass spectrometry. Results We identified 464 lipid species representing 18 lipid classes and revealed distinct stage-specific changes in lipid composition throughout nematode development. Triacylglycerols (TGs) dominated the lipid profile in the egg stage, suggesting a key role for them in energy storage at this early developmental stage. As N. brasiliensis develops, there was a conspicuous transition toward membrane-associated lipids, including glycerophospholipids (e.g. PE and PC) and ether-linked lipids, particularly in adult stages, indicating a shift toward host adaptation and membrane stabilisation. Conclusions We provide a comprehensive insight into the lipid composition and abundance of key free-living and parasitic stages of N. brasiliensis. This study provides lipidomic resources to underpin the detailed exploration of lipid biology in this model parasitic nematode. Graphical Abstract

The prevalence of metabolic dysfunction-associated steatohepatitis (MASH) is increasing at an alarming rate. To date, only one therapy has been provisionally approved for the treatment of MASH and liver fibrosis, and novel strategies are urgently needed. In addition, the frequent coexistence of MASH and type 2 diabetes has further intensified interest in devising comprehensive therapies to simultaneously tackle both diseases. We have recently shown that increasing hepatic and/or circulating levels of hexosaminidase A (HEXA), a lysosomal enzyme that remodels GM2 to GM3 gangliosides within lipid rafts, offers therapeutic benefits for metabolic dysfunction-associated steatotic liver disease (MASLD) and type 2 diabetes. Taking advantage of the MUP-uPA mouse model of MASH, including both wild-type (WT) mice with mild MASH and MUP-uPA mice with severe MASH and fibrosis, we show that biweekly treatment with a long-lasting HEXA-FC analog improves features of MASLD, including hepatic steatosis and hepatocyte ballooning, in mice with mild MASH, as well as glycemic control across both mouse models. Mechanistically, HEXA-FC enhances hepatic fatty acid oxidation and peripheral glucose disposal while not impacting endogenous glucose production. Together, these outcomes suggest that while HEXA-FC treatment may offer therapeutic benefits in mild MASH and insulin resistance, it is ineffective against severe MASH and liver fibrosis. NEW & NOTEWORTHY The prevalence of metabolic dysfunction-associated steatohepatitis (MASH) and type 2 diabetes is increasing. Here, we show that chronic FC-HEXA recombinant protein treatment reduces hepatic lipid accumulation and improves blood glucose control in mice with mild MASH and insulin resistance.

Current antidepressants have limitations due to insufficient efficacy and delay before improvement in symptoms. Polymorphisms of the serotonin transporter (5-HTT) gene have been linked to depression (when combined with stressful life events) and to altered response to selective serotonergic reuptake inhibitors. We have previously revealed the antidepressant-like properties of the iron chelator deferiprone in the 5-HTT knock-out (KO) mouse model of depression. Furthermore, deferiprone was found to alter neural activity in the prefrontal cortex of both wild-type (WT) and 5-HTT KO mice. In the current study, we examined the molecular effects of acute deferiprone treatment in the prefrontal cortex of both genotypes via phosphoproteomics. In WT mice treated with deferiprone, there were 22 differentially expressed phosphosites, with gene ontology analysis implicating cytoskeletal proteins. In 5-HTT KO mice treated with deferiprone, we found 33 differentially expressed phosphosites. Gene ontology analyses revealed phosphoproteins that were predominantly involved in synaptic and glutamatergic signalling. In a drug naive cohort, the analysis revealed 21 differentially expressed phosphosites in 5-HTT KO compared to WT mice. We confirmed the deferiprone-induced increase in Tyrosine hydroxylase serine 40 residue phosphorylation (pTH-Ser40) (initially revealed in our phosphoproteomics study) by western blots, with deferiprone increasing pTH-Ser40 expression in WT and 5-HTT KO mice. As glutamatergic and synaptic signalling are dysfunctional in 5-HTT KO mice (and are the target of fast-acting antidepressant drugs such as ketamine), these molecular effects may underpin deferiprone’s antidepressant-like properties. Furthermore, dopaminergic signalling may also be involved in deferiprone’s antidepressant-like properties.

BACKGROUND:Mass spectrometry-based quantitative proteomics has a demonstrated utility in increasing the diagnostic yield of mitochondrial disorders (MDs) and other rare diseases. However, for this technology to be widely adopted in routine clinical practice, it is crucial to accurately estimate delivery costs. Resource use and unit costs required to undertake a proteomics test were measured and categorized into consumables, equipment, and labor. Unit costs were aggregated to obtain a total cost per patient, reported in 2023 Australian dollars (AUD). Probabilistic and deterministic sensitivity analysis were conducted to evaluate parameter uncertainty and identify key cost drivers. RESULTS:The mean cost of a proteomics test was $897 (US$ 607) per patient (95% CI: $734-$1,111). Labor comprised 53% of the total costs. At $342 (US$ 228) per patient, liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) was the most expensive non-salary component. An integrated analysis pipeline where all the standard analysis are performed automatically, as well as discounts or subsidized LC-MS/MS equipment or consumables can lower the cost per test. CONCLUSIONS:Proteomics testing provide a lower-cost option and wider application compared to respiratory chain enzymology for mitochondrial disorders and potentially other functional assays in Australia. Our analysis suggests that streamlining and automating workflows can reduce labor costs. Using PBMC samples may be a cheaper and more efficient alternative to generating fibroblasts, although their use has not been extensively tested yet. Use of fibroblasts could potentially lower costs when fibroblasts are already available by avoiding the expense of isolating PBMCs. A joint evaluation of the health and economic implications of proteomics is now needed to support its introduction to routine clinical care.

The biological effects of electromagnetic field (EMF) irradiation in the terahertz (THz) range remain ambiguous, despite numerous studies that have been conducted. In this paper, the metabolic response of Escherichia coli K 12 to EMF irradiation was examined using a 1.0 W m-2 incident synchrotron source (SS) in the range of 0.5-18.0 THz for over 90 min of continuous exposure at 25 degrees C. This continuous SS THz exposure induced periodic decreases in the cell growth after 10, 20, and 40 min of exposure compared to a time-matched control; however, the number of viable cells thereafter grew. The physiological status of treated cells immediately after exposure was assessed by using the direct plate counting technique and electron microscopy. Analysis of scanning electron microscopy (SEM) and high-resolution cryogenic transmission electron (cryo-TEM) micrographs showed that approximately 20% of the SS THz-exposed E. coli cells exhibited a deformed outer membrane, membrane perturbations, and leakage of cytosol. The proteome changes in E. coli cells after 18 h postexposure were associated with cellular response to plasma membrane regulation including phospholipid biosynthetic process and osmotic stress. The results of this study highlighted that E. coli cells can promptly activate the fundamental mechanisms in response to prolonged exposure to THz radiation that are evolutionarily developed to withstand other environmental stressors.