Relative Protein Content Research Articles

Mucin Colocalizes with Influenza Virus and Preserves Infectivity in Deposited Model Respiratory Droplets.

The stability of influenza virus in respiratory particles varies with relative humidity (RH) and protein content. This study investigated the decay, or loss of infectivity, of influenza A virus (IAV) in 1-μL respiratory droplets deposited on a surface with varying concentrations of mucin, one of the most abundant proteins in respiratory mucus, and examined the localization of virions within droplets. IAV remained stable at 0.1% and 0.5% mucin in phosphate-buffered saline (PBS) over 4 h at 20%, 50%, and 80% RH, with a maximum decay of 1.2 log10/mL. In contrast, in pure PBS droplets, the virus decayed by at least 2.6 log10/mL after 4 h at 50% and 80% RH. Mucin's protective effect was independent of its concentration, except at 80% RH after 4 h. Confocal microscopy of the particles revealed that at 20% and 50% RH, mucin led to thicker coffee rings and dendritic patterns where virions colocalized with mucin. At 80% RH, no morphological difference was observed between PBS-only and mucin-containing droplets, but virions still colocalized with mucin in the center of droplets with 0.5% mucin. Analysis by digital droplet PCR showed that mucin helped maintain virus integrity. To our knowledge, this is the first study to localize influenza virus in model respiratory droplets. The results suggest that mucin's colocalization with virions in droplets may protect the virus from environmental stressors, enhancing its stability.

FTIR microscopic study on biomolecular alterations in placental tissues of intrahepatic cholestasis of pregnancy.

Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific liver disorder that typically leads to severe pregnancy outcomes. Although genetic, endocrine, and environmental factors are involved in the etiology of ICP, the role of metabolic disorders remains unclear. Here we report an examination of the biomolecular alterations in placental tissues of women with ICP and healthy pregnant women at a molecular level. By integrating FTIR microspectroscopy with advanced multivariate statistical analysis and semiquantitative methods, including PCA, OPLS-DA and peak area ratio calculations, the biomolecular alterations were revealed. Specifically, alterations in lipid conformations and increased lipid acyl chain unsaturation in the ICP group were associated with bile acids participating in the regulation of lipid metabolism. Additionally, a reduction in the relative α-helix content of proteins compared to β-sheet structures was associated with changes in apoptosis-related proteins. Furthermore, the nucleic acid content relative to lipids and proteins was elevated in the ICP group. Our study underscores the potential of FTIR microspectroscopy as a powerful tool for investigating the underlying biochemical mechanisms related to ICP.

Evaluating TAB2, IKBKB, and IKBKG Gene Polymorphisms and Serum Protein Levels and Their Association with Age-Related Macular Degeneration and Its Treatment Efficiency.

Background and Objectives: Age-related macular degeneration (AMD) is the leading cause of blindness, affecting millions worldwide. Its pathogenesis involves the death of the retinal pigment epithelium (RPE), followed by photoreceptor degeneration. Although AMD is multifactorial, various genetic markers are strongly associated with the disease and may serve as biomarkers for evaluating treatment efficacy. This study investigates TAB2 rs237025, IKBKB rs13278372, and IKBKG rs2472395 variants and their respective serum protein concentrations in relation to AMD occurrence and exudative AMD treatment response to anti-VEGF treatment. Materials and Methods: The case-control study involved 961 individuals, and they were divided into three groups: control, early AMD, and exudative AM patients. Genotyping of selected SNPs were conducted using a real-time polymerase chain reaction method (RT-PCR). Based on the clinical OCT and BCVA data, patients with exudative AMD were categorized into one of two groups: responders and non-responders. The data obtained were analyzed using the "IBM SPSS Statistics 29.0" software program. Results: Our study revealed that TAB2 rs237025 allele A was identified as a risk factor for early and exudative AMD development. The same associations remained only in females with exudative AMD but not in males, suggesting gender-specific pathogenetic pathways in exudative AMD. Analysis of IKBKB rs13278372 or serum IKBKB protein associations with early or exudative AMD occurrence in the Lithuanian population revealed no significant associations. On the other hand, we found that each A allele of IKBKB rs13278372 was associated with a worse response to anti-VEGF treatment (OR = 0.347; 95% CI: 0.145-0.961; p = 0.041). These results suggest a potential marker for future studies evaluating anti-VEGF treatment for exudative AMD patients. IKBKG rs2472395 was a protective variant for early AMD in males and for exudative AMD in females only. Also, IKBKG protein concentration was lower in exudative AMD relative to the control group (median (IQR): 0.442 (0.152) vs. 0.538 (0.337), p = 0.015). Moreover, exudative AMD patients who carry the GG genotype of IKBKG rs2472394 exhibited significantly reduced serum IKBKG concentrations compared to the controls (median (IQR): 0.434 (0.199) vs. 0.603 (0.335), p = 0.012), leading to the hypothesis that the IKBKG rs2472394 variant might play a role in protein concentration differences and exudative AMD development. Conclusions: Our study identified the TAB2 rs237025 allele A as a significant risk factor for both early and exudative AMD, with gender-specific associations observed in females with exudative AMD, suggesting distinct pathogenetic pathways. While IKBKB rs13278372 and serum IKBKB protein levels showed no significant association with AMD development, the A allele of IKBKB rs13278372 was associated with a worse response to anti-VEGF treatment, indicating its potential as a marker for treatment outcomes. Additionally, the IKBKG rs2472395 variant was found to be protective for early AMD in males and exudative AMD in females, and lower IKBKG protein levels were associated with exudative AMD, particularly in patients with the GG genotype of IKBKG rs2472394, suggesting its role in protein concentration and disease progression. These findings highlight genetic markers that may contribute to AMD pathogenesis and treatment response.

How different successive elaboration methods affect Hermetia illucens meals? Macronutrients, in vitro protein digestibility, oxidative status and hygienic-sanitary quality

Abstract Insects are an emerging protein alternative in aquaculture. To obtain an easy-to-handle feed with an improved nutritional quality, longer shelf life and better hygienic-sanitary properties, the insects must go through a multi-step process including sacrifice, drying and defatting. This work aims to evaluate the effect of the interaction of different successive methods of processing of Hermetia illucens larvae proximate composition, digestibility, and microbial loading. The examined methods were slaughtering (blanching or freezing), drying (oven drying or freeze drying) and defatting (mechanical press or supercritical CO2 extraction). Our results showed that the combination of freeze-drying and mechanical press defatting resulted in the highest residual fat content (), that could negatively affect the relative protein content of the meals. Despite this, with respect to in vitro amino acid digestibility, the least favorable results were obtained from the samples dried by lyophilization and defatted by supercritical CO2 extraction (), independently of the slaughter method. Regarding the oxidative status, the slaughter by freezing conserved a higher level of antioxidant defenses versus the blanching method (), although only the freeze drying method ensured a lower level of lipid oxidation in this type of sacrificing method. The applied processes resulted in a decrease in the presence of pathogenic bacteria compared to the raw insect in all examined groups. The mesophilic aerobic microorganism content was heavily affected by the temperature applied (), while the enterobacteria were also affected by the lyophilization, in addition to temperature. Salmonella spp was eliminated from all meals, except for the combination of freezing (sacrifice), freeze drying (drying) and supercritical CO2 extraction (defatting). Sulfite-reducing clostridia were not present in any of the prepared insect meals.

Changes in photoperiod during the dry period impact colostrum production in Holstein and Jersey cows.

Multiparous Holstein cows exposed to short-day photoperiod (SDPP) of 8 h of light per day during their dry period produced up to 3.2 kg more milk per day compared with cows exposed to long-day photoperiod (LDPP) of 16 h of light per day; it is unknown if a similar response would be observed for Jersey cow milk production. The objective of this study was to determine the effect of photoperiod during the dry period on subsequent colostrum and milk production in Holstein and Jersey cattle. Holstein and Jersey cows (n = 33) were dried off 60 d before their due date and randomly assigned to SDPP (Holstein, n = 9; Jersey, n = 8) or LDPP (Holstein, n = 8; Jersey, n = 8) until calving. Cows were weighed at the time of enrollment (d 0) and were housed in an enclosed barn at 20°C and exposed to 250 to 450 lx during periods of light and <10 lx during periods of darkness. At calving, colostrum volume was weighed and tested for relative protein concentration with a Brix refractometer and a sample was collected for component analysis (fat, protein, lactose, SNF) via infrared spectroscopy, as well as IgA, IgG, IgG1, IgM, lactoferrin, and SCS analysis. After calving, cows were returned to the freestall barn and exposed to ambient photoperiod and temperature. Milk production data were collected for 15 wk postcalving. Data were analyzed using PROC MIXED in SAS (SAS 9.4; SAS Institute Inc., Cary, NC) with treatment, breed, and d 0 weight as fixed effects. PROC MIXED with repeated measures was used to evaluate the relationship of day length and breed with mature milk volume, fat, and protein production. Random effects included replicate, lactation number, genetic inbreeding percentage, previous lactation mature equivalent 305-d protein production, and calf sex. For colostrum, Brix score, colostral protein, fat, IgA, and IgM were increased in Jersey cows compared with Holstein cows. Total colostrum weight, SNF, lactose, lactoferrin, IgG, IgG1, and SCS did not differ by breed or treatment. Postcalving, ECM production was increased in Holstein cows compared with Jersey cows but unaffected by photoperiod treatment. Conversely, milk protein percentage was increased for Jersey cows relative to Holstein cows but was unaffected by photoperiod treatment. Milk fat increased in LDPP Holstein cows compared with SDPP Jersey cows during the first week of lactation, which is likely due to the transition from colostrum to mature milk production. Overall, photoperiod did not affect colostrum production, but differences by breed were detected. Photoperiod during the dry period did not affect mature milk production or protein, but milk fat percentage was affected by photoperiod × breed. Therefore, altered lighting during the dry period does not unfavorably affect colostrum or milk production in Jersey or Holstein cows.

Orange peel biochar: An effective amendment to improve the maize resilience by regulating the soil enzymatic activities, nutrient uptake, and ionic homeostasis under salinity stress

The conversion of organic waste into biochar offers innovative solutions for soil remediation. Globally, salinization is a major threat to agricultural productivity, significantly contributing to soil degradation. The application of orange peel (OP) waste could provide a better solution for sustainable crop productivity. The study investigated the influence of OP and orange peel biochar (OPB) at varying doses (0.5 %, 1 %, and 2 %) on soil enzymes and maize growth under different salinity levels (1.66, 4, and 8 dS m–¹). The results indicate that OPB was more effective than OP, with promising outcomes even at lower rates. At 8 dS m–1, 1 % OPB significantly enhanced shoot length (30.7 %), root length (21.9 %), grain yield (3-fold), relative water content (14.2 %), protein content (32.1 %), chlorophyll value (51.9 %), carotenoid content (31.5 %), and grain N, P, and K contents (27.5 %, 54.0 %, and 17.8 %), while significantly reducing Na contents in roots (16.6 %) and shoots (27.8 %) compared to the control. Furthermore, 1 % OPB was an optimal rate for enhancing soil enzymatic activities, including urease (55.4 %), acid phosphatase (37.8 %), and alkaline phosphatase (47.4 %), outperforming the control at 8 dS m–1. The structural equation model (SEM) revealed that improving soil enzymatic activities was the main mechanism accelerated by OPB to improve the maize growth and yield. Adding 1 % OPB to soil is an efficient and effective approach for alleviating the toxic effects of salinization to improve maize growth, physiology, and yield. These findings will advance OP recycling as a cost-effective solution for sustainable agriculture and mitigating salinity stress.

A Proteomics-Based Approach for Prediction of Different Cardiovascular Diseases and Dementia.

Many studies have explored whether individual plasma protein biomarkers improve cardiovascular disease risk prediction. We sought to investigate the use of a plasma proteomics-based approach in predicting different cardiovascular outcomes. Among 51 859 UK Biobank participants (mean age, 56.7 years; 45.5% male) without cardiovascular disease and with proteomics measurements, we examined the primary composite outcome of fatal and nonfatal coronary heart disease, stroke, or heart failure (major adverse cardiovascular events), as well as additional secondary cardiovascular outcomes. An exposome-wide association study was conducted using relative protein concentrations, adjusted for a range of classic, demographic, and lifestyle risk factors. A prediction model using only age, sex, and protein markers (protein model) was developed using a least absolute shrinkage and selection operator-regularized approach (derivation: 80% of cohort) and validated using split-sample testing (20% of cohort). Their performance was assessed by comparing calibration, net reclassification index, and c statistic with the PREVENT (Predicting Risk of CVD Events) risk score. Over a median 13.6 years of follow-up, 4857 participants experienced first major adverse cardiovascular events. After adjustment, the proteins most strongly associated with major adverse cardiovascular events included NT-proBNP (N-terminal pro B-type natriuretic peptide; hazard ratio [HR], 1.68 per SD increase), proADM (pro-adrenomedullin; HR, 1.60), GDF-15 (growth differentiation factor-15; HR, 1.47), WFDC2 (WAP four-disulfide core domain protein 2; HR, 1.46), and IGFBP4 (insulin-like growth factor-binding protein 4; HR, 1.41). In total, 222 separate proteins were predictors of all outcomes of interest in the protein model, and 86 were selected for the primary outcome specifically. In the validation cohort, compared with the PREVENT risk factor model, the protein model improved calibration, net reclassification (net reclassification index +0.09), and c statistic for major adverse cardiovascular events (+0.051). The protein model also improved the prediction of other outcomes, including ASCVD (c statistic +0.035), myocardial infarction (+0.023), stroke (+0.024), aortic stenosis (+0.015), heart failure (+0.060), abdominal aortic aneurysm (+0.024), and dementia (+0.068). Measurement of targeted protein biomarkers produced superior prediction of aggregated and disaggregated cardiovascular events. This study represents an important proof of concept for the application of targeted proteomics in predicting a range of cardiovascular outcomes.

Covalent Attachment of Functional Proteins to Microfiber Surfaces via a General Strategy for Site-Selective Tetrazine Ligation.

Surface modification of materials with proteins has various biological applications, and hence the methodology for surface modification needs to accommodate a wide range of proteins that differ in structure, size, and function. Presented here is a methodology that uses the Affinity Bioorthogonal Chemistry (ABC) tag, 3-(2-pyridyl)-6-methyltetrazine (PyTz), for the site-selective modification and purification of proteins and subsequent attachment of the protein to trans-cyclooctene (TCO)-functionalized hydrogel microfibers. This method of surface modification is shown to maintain the functionality of the protein after conjugation with proteins of varying size and functionalities, namely, HaloTag, NanoLuc luciferase (NanoLuc), and fibronectin type III domains 9-10 (FNIII 9-10). The method also supports surface modification with multiple proteins, which is shown by the simultaneous conjugation of HaloTag and NanoLuc on the microfiber surface. The ability to control the relative concentrations of multiple proteins presented on the surface is shown with the use of HaloTag and superfolder GFP (sfGFP). This application of the ABC-tagging methodology expands on existing surface modification methods and provides flexibility in the site-selective protein conjugation methods used along with the rapid kinetics of tetrazine ligation.

Identification of salt tolerance of different Chinese kale cultivars under NaCl stress at seedling stage

In order to understand the salt tolerance of different Chinese kale varieties at seedling stage and screen out the varieties with strong salt tolerance at seedling stage, the seeds of 26 Chinese kale varieties were used as materials. The growth indexes, physiological indexes, seedling rate and salt damage index of different Chinese kale varieties were compared and cluster analyzed by means of tissue culture. The results showed that: (1) under 160 mmol∙L−1 NaCl stress, the different Chinese kale varieties at growth indexes, physiological indexes, seedling rate and salt damage index were significantly different, the relative seedling rate of R4, R14 and R32 reached 100.00%, and the salt damage index was the lowest, which was 20.00%; The correlation analysis showed that the relative seedling rate, the relative content of soluble protein, the relative content of soluble sugar and the relative value of Leaf SPAD were all negatively correlated with the salt damage index, and the relative conductivity and the relative content of malondialdehyde were all positively correlated with the salt damage index. (2) The average value of 14 salt tolerance indexes was used to cluster analyze the salt tolerance of 26 Chinese kale varieties. The tested materials were divided into four categories: high salt tolerance, moderate salt tolerance, low salt tolerance and salt sensitivity. R14, R25, R30 and R32 were highly salt tolerant varieties and R24 was salt sensitive varieties, which laid a foundation for further breeding and genetic research of salt tolerant varieties.

Label-Free Optical Biopsy Reveals Biomolecular and Morphological Features of Diabetic Kidney Tissue in 2D and 3D.

Kidney disease, the ninth leading cause of death in the United States, has one of the poorest diagnostic efficiencies of only 10%1. Conventional diagnostic methods often rely on light microscopy analysis of 2D fixed tissue sections with limited molecular insight compared to omics studies. Targeting multiple features in a biopsy using molecular or chemical reagents can enhance molecular phenotyping but are limited by overlap of their spatial and chromatic properties, variations in quality of the products, limited multimodal nature and need additional tissue processing. To overcome these limitations and increase the breadth of molecular information available from tissue without an impact on routine diagnostic workup, we implemented label-free imaging modalities including stimulated Raman scattering (SRS) microscopy, second harmonic generation (SHG), and two photon fluorescence (TPF) into a single microscopy setup. We visualized and identified morphological, structural, lipidomic, and metabolic biomarkers of control and diabetic human kidney biopsy samples in 2D and 3D at a subcellular resolution. The label-free biomarkers, including collagen fiber morphology, mesangial-glomerular fractional volume, lipid saturation, redox status, and relative lipid and protein concentrations in the form of Stimulated Raman Histology (SRH), illustrate distinct features in kidney disease tissues not previously appreciated. The same tissue section can be used for routine diagnostic work up thus enhancing the power of cliniopathological insights obtainable without compromising already limited tissue. The additional multimodal biomarkers and metrics are broadly applicable and deepen our understanding of the progression of kidney diseases by integrating lipidomic, fibrotic, and metabolic data.

Compound extreme heat and drought stress alter the spatial gradients of protein and starch in wheat grains

The spatial gradients of protein and starch in wheat grains affected the grain milling characteristics and flour utilization. The increase in compound heat stress and drought stress (HDS) due to global climate change threatens wheat grain yield and quality parameters, but the impacts of extreme climate events on the gradients of protein and starch in wheat grains remain unclear. In this study, two-year, environment-controlled experiments with four heat stress levels (17/27, 21/31, 25/35, and 29/39°C) and three drought stress levels (30 %, 55 %, and 75 % field capacity) were conducted to investigate the effects of HDS on the gradients of protein and starch concentrations within the five grain layers. The results showed that HDS resulted in significantly greater protein concentrations, while resulting in lower starch concentrations in the wheat grain layers. Among the five layers, the endosperm layers exhibited the greatest increase in protein concentration under HDS, but the starch concentration under HDS decreased in the order of husk > aleurone > endosperm layer. HDS unevenly altered the protein and starch concentrations of the five grain layers. There was significant linear relationship between relative protein and starch concentration with accumulated heat degree days (AHDD). With a 1°C·d increase in AHDD, the protein concentration in the five grain layers increased by 1.17–2.19 %, while the starch concentration decreased by 0.62–0.90 %, depending on the drought stress levels. A significant linear relationship was also observed between the relative protein and starch concentrations and evapotranspiration (ET). A 1 mm increase in ET led to a protein concentration decrease of 0.58–0.76 % in P1-P5, with a corresponding starch concentration increase of 0.28–0.46 %, depending on cultivar and treatment stages. Our results indicate that HDS significantly impacts the grain quality parameters for the flour milling process and human diet and will provide important insights into adapting wheat quality to climate change.

Soft X-ray spectromicroscopy of human fibroblasts with impaired sialin function.

Salla disease (SD) is a lysosomal storage disease where free sialic acid (SA) accumulates in lysosomes due to the impaired function of a membrane protein, sialin. Synchrotron radiation-based scanning transmission soft X-ray spectromicroscopy (STXM) was used to analyze both SD patients' fibroblasts and normal human dermal fibroblasts (NHDF) from healthy controls. Both cell lines were also cultured with N-acetyl-d-mannosamine monohydrate (ManNAc) to see if it increased SA concentration in the cells. The STXM technique was chosen to simultaneously observe the morphological and chemical changes in cells. It was observed that free SA did not remain in the lysosomes during the sample processing, leaving empty vacuoles to the fibroblasts. The total cytosol and entire cell spectra, however, showed systematic differences between the SD and NHDF samples, indicating changes in the relative macromolecular concentrations of the cells. The NHDF cell lines contained a higher relative protein concentration compared to the SD cell lines, and the addition of ManNAc increased the relative protein concentration in both cell lines. In this study, two sample preparation methods were compared, resin-embedded thin sections and cells grown directly on sample analysis grids. While the samples grown on the grids exhibited clean, well-resolved spectra not masked by embedding resin, the low penetration depth of soft X-rays hindered the analysis to only the thin region of the microfilaments away from the thick nucleus.

Relationships between HDL subpopulation proteome and HDL function in overweight/obese people with and without coronary heart disease

Background and aimsThe structure-function relationships of high-density lipoprotein (HDL) subpopulations are not well understood. Our aim was to examine the interrelationships between HDL particle proteome and HDL functionality in subjects with and without coronary heart disease (CHD). MethodsWe isolated 5 different HDL subpopulations based on charge, size, and apolipoprotein A1 (APOA1) content from the plasma of 33 overweight/obese CHD patients and 33 age-and body mass index (BMI)-matched CHD-free subjects. We measured the relative molar concentration of HDL-associated proteins by liquid chromatography tandem mass spectrometry (LC-MS/MS) and assessed particle functionality. ResultsWe quantified 110 proteins associated with the 5 APOA1-containing HDL subpopulations. The relative molar concentration of these proteins spanned five orders of magnitude. Only 10 proteins were present in >1% while 73 were present in <0.1% concentration. Only 6 of the 10 most abundant proteins were apolipoproteins. Interestingly, the largest (α-1) and the smallest (preβ-1) HDL particles contained the most diverse proteomes. The protein composition of each HDL subpopulation was altered in CHD cases as compared to controls with the most prominent differences in preβ-1 and α-1 particles. APOA2 concentration was positively correlated with preβ-1 particle functionality (ABCA1-CEC/mg APOA1 in preβ-1) (R2 = 0.42, p = 0.005), while APOE concentration was inversely correlated with large-HDL particle functionality (SRBI-CEC/mg APOA1 in α-1+α-2) (R2 = 0.18, p = 0.01). ConclusionsThe protein composition of the different HDL subpopulations was altered differentially in CHD patients. The functionality of the small and large HDL particles correlated with the protein content of APOA2 and APOE, respectively. Our data indicate that distinct particle subspecies and specific particle associated proteins provide new information about the role of HDL in CHD.

Evaluation of Foliar Application of &lt;i&gt;Elusine indica&lt;/i&gt; Extract on Growth, Photosynthesis, and Osmoprotectant Contents in Maize under Drought Stress

To investigate the effects of foliar application of different concentrations of &lt;i&gt;Elusine indica&lt;/i&gt; extract (EIE) on growth, photosynthesis, and osmoprotectant contents in maize under drought stress. The weed powder was extracted using methanol, followed by a solid-liquid extraction procedure. Plants were sprayed with three different concentrations of EIE at 1, 3, and 5 g/L and morphological parameters, chlorophyll, relative water content (RWC), soluble sugar, proline, protein, glutathione (GSH), and malondialdehyde (MDA) contents were determined. The results showed that drought stress led to a decline in morphological characteristics, RWC and soluble sugar and increased proline, protein, GSH, and MDA contents. However, foliar application of EIE significantly improved plant height, fresh and dry weight, chlorophyll content, RWC, soluble sugar, and GSH, while the proline level was diminished compared to drought treatment. Soluble sugar showed a significant positive correlation with fresh and dry weight (&lt;i&gt;r&lt;/i&gt; = 0.742 and 0.783, &lt;i&gt;p&lt;/i&gt; &lt; 0.01) and a strong negative correlation with MDA (&lt;i&gt;r&lt;/i&gt; = -0.459, &lt;i&gt;p&lt;/i&gt; &lt; 0.05). Therefore, this result indicated that EIE can be used as an inexpensive and environmentally friendly biostimulant to help plants enhance tolerance to drought.

Identification of brain-enriched proteins in CSF as biomarkers of relapsing remitting multiple sclerosis

BackgroundMultiple sclerosis (MS) is a clinically and biologically heterogenous disease with currently unpredictable progression and relapse. After the development and success of neurofilament as a cerebrospinal fluid (CSF) biomarker, there is reinvigorated interest in identifying other markers of or contributors to disease. The objective of this study is to probe the predictive potential of a panel of brain-enriched proteins on MS disease progression and subtype.MethodsThis study includes 40 individuals with MS and 14 headache controls. The MS cohort consists of 20 relapsing remitting (RR) and 20 primary progressive (PP) patients. The CSF of all individuals was analyzed for 63 brain enriched proteins using a method of liquid-chromatography tandem mass spectrometry. Wilcoxon rank sum test, Kruskal-Wallis one-way ANOVA, logistic regression, and Pearson correlation were used to refine the list of candidates by comparing relative protein concentrations as well as relation to known imaging and molecular biomarkers.ResultsWe report 30 proteins with some relevance to disease, clinical subtype, or severity. Strikingly, we observed widespread protein depletion in the disease CSF as compared to control. We identified numerous markers of relapsing disease, including KLK6 (kallikrein 6, OR = 0.367, p < 0.05), which may be driven by active disease as defined by MRI enhancing lesions. Other oligodendrocyte-enriched proteins also appeared at reduced levels in relapsing disease, namely CNDP1 (carnosine dipeptidase 1), LINGO1 (leucine rich repeat and Immunoglobin-like domain-containing protein 1), MAG (myelin associated glycoprotein), and MOG (myelin oligodendrocyte glycoprotein). Finally, we identified three proteins—CNDP1, APLP1 (amyloid beta precursor like protein 1), and OLFM1 (olfactomedin 1)—that were statistically different in relapsing vs. progressive disease raising the potential for use as an early biomarker to discriminate clinical subtype.ConclusionsWe illustrate the utility of targeted mass spectrometry in generating potential targets for future biomarker studies and highlight reductions in brain-enriched proteins as markers of the relapsing remitting disease stage.

P19 Defining the mechanism of skin barrier restoration by Lactobacillus rhamnosus

Abstract Introduction and aims We aim to investigate the lysate of Lactobacillus rhamnosus GG (LGG), a bacterium previously established as a gut probiotic, and to examine its postulated ability to enhance and restore human skin barrier function. Compromised skin barriers are implicated in various skin conditions including atopic dermatitis. Furthermore, skin barrier function decreases with age leading to dryness and sensitivity. Previous research has shown promising effects of LGG lysates on keratinocytes, including increasing transepidermal electrical resistance, increasing tight junction protein expression, cell migration promotion and mitigation of Staphylococcus aureus toxicity. This study aims to take a closer look at the molecular mechanisms behind the effect of LGG lysates on the skin, including terminal differentiation, lamin dispersal and nuclear degradation in keratinocytes. Methods Normal human epithelial keratinocytes and rat epithelial keratinocytes were grown and treated with the LGG lysate. Western blot analysis was used to determine the relative concentrations of tight junction proteins and other differentiation markers. A phosphoproteomic study was performed to identify key pathways activated by the LGG lysate. A Staphylococcus epidermidis lysate control was used to filter out any nonspecific bacterial effects. Immunostaining of nuclear proteins was used to visualize the effects of LGG lysate on lamin dispersal and nuclear degradation. short hairpin RNA filaggrin knockdown normal human keratinocytes were developed to create a ‘barrier impaired’ model to assess ability of the lysate to ‘rescue’ the skin barrier. Results The LGG lysate was shown to increase tight junction protein expression in normal human keratinocytes and increase keratinocyte terminal differentiation markers. We present Results that show decreased tight junction protein expression in filaggrin knockdown cells. Conclusions These Results show the potential for LGG lysate to enhance and restore skin barrier function and begin to suggest potential mechanisms for this effect.

Comparison of Morphological, Physiological, and Related Gene Expression Responses to Drought Stress in Five Camellia vietnamensis Cultivars

The main production area of Camellia vietnamensis (C. vietnamensis) is in the low mountain and hilly areas of southern China. The low survival rate of seedlings caused by drought is one of the main obstacles restricting the development of the C. vietnamensis industry. An exploration of the key adaptation mechanism of C. vietnamensis to drought stress is important in order to improve its drought resistance. We conducted a study on the morphological, physiological, biochemical, and drought resistance-related genes of five C. vietnamensis cultivars grown in Hainan province under varying degrees of drought stress. The results indicate that drought stress can lead to a decrease in the relative water content and photosynthetic capacity of C. vietnamensis leaves. Compared with the control, the drought damage index, malondialdehyde, relative electrical conductivity, soluble protein, soluble sugar and proline contents of the five C. vietnamensis cultivars increased with drought-stress duration and degree. With increasing drought-stress intensity, the activity of antioxidant enzymes and the content of related metabolites (total polyphenols, total flavonoids, tea saponins) gradually increased, and the expression levels of phenylpropanoid pathway-related genes (Cv4CL1, CvCAD1, CvCAD2, CvPOX1, CvPOX2, CvPOX3) were upregulated. Based on the results of the drought tolerance coefficients, principal component analysis, and hierarchical cluster analysis, we classified five C. vietnamensis cultivars into drought-tolerant cultivars (‘Haida 1’); moderately drought-tolerant cultivars (‘Haida 4’ and ‘Wanhai 4’); and drought-sensitive cultivars (‘Wanhai 3’ and ‘Wanhai 1’). The results of this study provide a theoretical basis for the promotion and cultivation of C. vietnamensis and the selection of drought-resistant cultivars.

Treatment of seeds by cold ambient air plasma: combining impedance measurements with water sorption modeling to understand the impact of seed hydration

In this article, we focus on the plasma seed interaction and more specifically-on the feedback exerted by the seeds on the plasma properties. Dormant Arabidopsis seeds with different water contents (WC), namely 3%DW, 10%DW and 30%DW were exposed to cold ambient air plasma (C2AP) generated in a dielectric barrier device (DBD). It is found that increasing WC enhances the capacitive current of the DBD, generates a greater number of low energy streamers (characterized by current peaks lower than 10 mA) that preferentially interplay with the seeds. Since the resistive and capacitive components of the seeds modify the C2AP electrical properties, impedance measurements (also called LCRmetry) have been carried out to measure their main dielectric parameters before/after plasma exposure (seeds resistance, capacitance, complex relative permittivity, tangent loss and conductivity). It appears that WC significantly changes dielectric losses at low frequencies (<1 kHz) due to polarization relaxation of the polar molecules (i.e. water). LCRmetry further reveals that C2AP does not substantially alter seeds dielectric parameters, i.e. it neither adds or removes significant amounts of new materials, meaning that the relative starch, protein and lipid contents remain essentially unaffected. However, it cannot be discounted that some bulk properties of the Arabidopsis seeds may be modified, especially regarding their porosity. This characteristic could facilitate penetration of plasma-generated reactive oxygen species into the internal seed tissues, leading to the grafting of oxygenated groups. To corroborate this theory, water sorption isotherms have been achieved on Arabidopsis seeds and fitted with four thermodynamic models, including the Brunauer–Emmett–Teller model and the Generalized D’Arcy and Watt model. It is demonstrated that C2AP primarily strengthens water-seed affinity by modifying molecular interactions rather than changing the seed’s moisture layer. This occurs despite a potential decrease in the number of adsorption sites, indicating a significant increase in overall seed hydrophilicity after plasma treatment.

Synergistic detoxification efficiency and mechanism of triclocarban degradation by a bacterial consortium in the liver-gut-microbiota axis of zebrafish (Danio rerio)

Triclocarban (TCC), an emerging organic contaminant, poses a potential threat to human health with long-term exposure. Here, Rhodococcus rhodochrous BX2 and Pseudomonas sp. LY-1 were utilized to degrade TCC at environmental related concentrations for enhancing TCC biodegradation and investigating whether the toxicity of intermediate metabolites is lower than that of the parent compound. The results demonstrated that the bacterial consortium could degrade TCC by 82.0% within 7 days. The calculated 96 h LC50 for TCC, as well as its main degradation product 3,4-Dichloroaniline (DCA) were 0.134 mg/L and 1.318 mg/L respectively. Biodegradation also alleviated histopathological lesions induced by TCC in zebrafish liver and gut tissues. Liver transcriptome analysis revealed that biodegradation weakened differential expression of genes involved in disrupted immune regulation and lipid metabolism caused by TCC, verified through RT-qPCR analysis and measurement of related enzyme activities and protein contents. 16 S rRNA sequencing indicated that exposure to TCC led to gut microbial dysbiosis, which was efficiently improved through TCC biodegradation, resulting in decreased relative abundances of major pathogens. Overall, this study evaluated potential environmental risks associated with biodegradation of TCC and explored possible biodetoxification mechanisms, providing a theoretical foundation for efficient and harmless bioremediation of environmental pollutants.

Physio-Biochemical Insights into the Cold Resistance Variations among Nectarine (Prunus persica (L.) Batsch var. nectarina) Cultivars.

Cold stress occurs in late winter and early spring threatens greatly the nectarine industry. In this study, the semi-lethal low temperature (LT50) and thirteen cold resistance related parameters of five nectarine cultivars, including 'Nonglehong little princess' (LP), 'Luyou No. 5' (LY), 'Nonglehong No. 6' (NL), 'Zhongyou No. 20' (ZY) and 'Qiuhongzhu' (QH), were determined. Based on these parameters, they were categorized into high-(HR, including NL and LP), moderate-(MR, including QH) and low-cold resistant (LR, including ZY and LY) groups. The relative water (RW), proline (PRO), soluble sucrose (SS) and soluble protein (SP) contents, and superoxide dismutase (SOD) and peroxidase (POD) activities of HR cultivars were higher while their relative electronic conductivity (RE), malondialdehyde (MDA) and gibberellin acid (GA3) contents and catalase (CAT) activity were lower than other cultivars during natural overwintering. Redundancy analysis revealed that the lowest temperature in a day (LT) and LT50 significantly explains 69.8% and 10.9% of these physiological variables, respectively. Moreover, GA3 and indoleacetic acid (IAA) contents and CAT activity were positively correlated, while PRO, SS, ABA and RW contents were negatively correlated with both LT and LT50. Our study will be helpful in understanding the cold resistance variations of nectarine germplasm resources.