Changes In Protein Content Research Articles

Investigation of the Changes in Concentrations of Vitamin D-Binding Protein and Lactoferin in Plasma and Peritoneal Fluid of Patients with Endometriosis.

An evaluation of the association between the concentrations of vitamin D-binding protein and lactoferrin in the plasma and peritoneal fluid may facilitate the elucidation of molecular mechanisms in endometriosis. Vitamin D-binding protein and lactoferrin concentrations were measured by ELISA in plasma and peritoneal fluid samples from 95 women with suspected endometriosis as classified by laparoscopy into groups with (n = 59) and without endometriosis (n = 36). There were no differences (p > 0.05) in the plasma and peritoneal fluid concentrations of vitamin D-binding protein and lactoferrin between women with and without endometriosis. In women with endometriosis, there was a significant correlation between plasma and peritoneal fluid vitamin D-binding protein concentrations (r = 0.821; p = 0.000), but there was no correlation between lactoferrin concentrations in those compartments (r = 0.049; p > 0.05). Furthermore, in endometriosis, lactoferrin was found to correlate poorly with vitamin D-binding protein (r= -0.236; p > 0.05) in plasma, while in the peritoneal fluid, the correlation between those proteins was significant (r = 0.399; p = 0.002). The characteristic properties of vitamin D-binding protein and lactoferrin and the associations between their plasma and peritoneal fluid concentrations found in women with endometriosis may provide a novel panel of markers to identify high-risk patients in need of further diagnostic measures.

Using Plasma NfL and GFAP to Monitor Response to Anti-CD20 Treatment in MS (P1-3.010)

<h3>Objective:</h3> To compare changes in plasma concentrations of Neurofilament light (NfL) and Glial Fibrillary Acidic Protein (GFAP) in individuals with relapsing multiple sclerosis (RMS) vs progressive multiple sclerosis (PMS), treated with anti-CD20 immunotherapy (rituximab or ocrelizumab) <h3>Background:</h3> Blood biomarkers, especially NfL, can augment clinical and radiographic data in MS treatment monitoring. Few studies have investigated longitudinal change in NfL or GFAP in PMS patients taking anti-CD20s or compared changes in these biomarkers between RMS and PMS. <h3>Design/Methods:</h3> Subjects were selected by MS diagnosis, anti-CD20 treatment for at least 6 months, and presence of multiple blood samples in our Center’s Biorepository. Demographic/clinical information was extracted by chart review. SIMOA plasma assays of NfL and GFAP were conducted at baseline and follow-up (between 3 and 12 months) on Quanterix SR-X. Biomarker concentrations were log transformed. Summary statistics and longitudinal regression analyses, correcting for age, were generated. <h3>Results:</h3> 37 subjects and 67 samples were analyzed (n=7 had no follow-up samples between 3 and 12 months). 64.9% were female. RMS (n=19) had mean(SD) age 38.7(9.2). PMS (n=18) had mean age 56.1(10.4). Age-adjusted mean concentrations of NfL and GFAP did not differ significantly between RMS and PMS at baseline. Geometric mean NfL declined between baseline (6.8, 95%CI 4.2–11.0) and follow-up (4.9, 95%CI 3.3–7.3) in RMS but stayed stable in PMS. Geometric mean GFAP increased between baseline (52.4, 95%CI 35.2–77.9) and follow-up (60.8, 95%CI 40.4–91.4) in PMS but stayed stable in RMS. The changes for both NfL and GFAP differed statistically significantly between MS types. <h3>Conclusions:</h3> With anti-CD20 treatment, plasma NfL levels decreased in RMS by 27.6% while staying stable in PMS; GFAP levels increased in PMS by 16% while remaining stable in RMS. Results correlate with clinical evidence showing greater benefit of anti-CD20 immunotherapy in RMS compared to PMS. Future studies will include more subjects, longer observation, and different immunotherapies. <b>Disclosure:</b> Dr. Gross has received personal compensation in the range of $5,000-$9,999 for serving as an Expert Witness for AP Expert Group. The institution of Stefan Sillau has received research support from Alzheimer’s Association. The institution of Stefan Sillau has received research support from Hewitt Family Foundation; State of Colorado. The institution of Stefan Sillau has received research support from PCORI. The institution of Stefan Sillau has received research support from NINR. The institution of Stefan Sillau has received research support from Michael J. Fox Foundation. The institution of Stefan Sillau has received research support from Department of Defense. The institution of Stefan Sillau has received research support from Colorado Department of Public Health and Environment. The institution of Stefan Sillau has received research support from Benign Essential Blepharospasm Research Foundation. Stefan Sillau has a non-compensated relationship as a Statistician with Novartis that is relevant to AAN interests or activities. Stefan Sillau has a non-compensated relationship as a Statistician with Biogen that is relevant to AAN interests or activities. Sean Selva has nothing to disclose. Mrs. Ritchie has nothing to disclose.

Forage quantity and protein concentration changes across a forest-savanna gradient with management implications for white-tailed deer

White-tailed deer (Odocoileus virginianus) hunting is an important economic activity associated with the management of forests and rangelands in the USA, with over $12.9 billion dollars of related annual expenditures. Reducing tree cover through thinning and prescribed fire both have the potential to increase the quantity and quality of deer forage. We evaluated the long-term impacts of eight different combinations of fire return intervals and tree harvest on forage productivity and protein content of the forage. Based on management regime, study units ranged from savanna to closed-canopy forest. Aboveground net primary production (ANPP) of six functional groups (grass, panicum, forb, legume, woody, sedge) of understory vegetation was measured in October 2019 and 2020 using destructive sampling. Samples for foliar crude protein (CP) concentration were collected in spring, summer, and fall of 2020. Total understory ANPP ranged from 2.9 to 466.3 g m−2 and was up to 566% greater in savanna systems maintained by frequent fire (return interval of three years or less) than in non-burned forest treatments. Annual burning resulted in ANPP dominated by herbaceous plants composed mostly of fire-tolerant grasses (e.g., Andropogon gerardii, Schizachyrium scoparium). Longer fire return intervals or no fire resulted in roughly equal ANPP from understory woody and herbaceous species. Crude protein concentrations were up to 45.7% greater in the woodland and forest units than in the savanna units for seven of the eleven species sampled. The greater CP in the forests was most noticeable in the summer when deer needs for quality forage are substantial. Increased protein concentrations of understory species in the forests, but greater ANPP in the savannas indicate that managing for a mix of savanna and woodland could be ideal for balancing forage quantity with increased forage protein.

Effects of functional oil blend on performance, blood metabolites, organ biometry and intestinal morphometry in nursery piglets

This study was conducted to assess the effects of functional oil (FO) blend on performance, blood metabolites, organ biometry and intestinal morphometry in piglets. A total of 128 crossbreed piglets (Landrace × Large White, 64 uncastrated males and 64 females, 21 d of age, and 6.79 ± 1.76 kg BW) were allocated in a randomized complete block design with two dietary treatments: a FO-free (FOF) diet or a diet based on added FO (1,500 mg/kg of diet with castor oil plus cashew nutshell oil). Piglets fed FO showed higher (p ≤ 0.05) average daily feed intake, daily body weight gain and final body weight after 23 d of study. For the total period, piglets fed FO showed greater (p = 0.007) feed conversion ratio. On d 23, higher serum total protein (p = 0.026) and globulin (p = 0.050) concentration, lower liver (p = 0.042) and stomach (p = 0.074) weight, and greater (p = 0.082) villi height (VH) in duodenum were observed in piglets fed FO. Nonetheless, piglets fed FOF showed greater (p = 0.054) ileal VH, but greater (p = 0.004) crypt depth (CD) in jejunum. Piglets fed FO showed higher VH to CD ratio in jejunum (p = 0.068) and duodenum (p = 0.074) on d 23 and 37, respectively. Based on the results, FO blend improved the performance of weaned piglets; however, it negatively affected the feed conversion ratio in the total period. Moreover, FO blend promoted changes in total protein concentrations and improvements in digestive and absorptive capacity assessed through VH to CD ratio, with a significant reduction in organs.

Spatiotemporal Live-Cell Analysis of Photoreceptor Outer Segment Membrane Ingestion by the Retinal Pigment Epithelium Reveals Actin-Regulated Scission.

The photoreceptor outer segment (OS) is the phototransductive organelle in the vertebrate retina. OS tips are regularly ingested and degraded by the adjacent retinal pigment epithelium (RPE), offsetting the addition of new disk membrane at the base of the OS. This catabolic role of the RPE is essential for photoreceptor health, with defects in ingestion or degradation underlying different forms of retinal degeneration and blindness. Although proteins required for OS tip ingestion have been identified, spatiotemporal analysis of the ingestion process in live RPE cells is lacking; hence, the literature reflects no common understanding of the cellular mechanisms that affect ingestion. We imaged live RPE cells from mice (both sexes) to elucidate the ingestion events in real time. Our imaging revealed roles for f-actin dynamics and specific dynamic localizations of two BAR (Bin-Amphiphysin-Rvs) proteins, FBP17 and AMPH1-BAR, in shaping the RPE apical membrane as it surrounds the OS tip. Completion of ingestion was observed to occur by scission of the OS tip from the remainder of the OS, with a transient concentration of f-actin forming around the site of imminent scission. Actin dynamics were also required for regulating the size of the ingested OS tip, and the time course of the overall ingestion process. The size of the ingested tip is consistent with the term "phagocytosis." However, phagocytosis usually refers to engulfment of an entire particle or cell, whereas our observations of OS tip scission indicate a process that is more specifically described as "trogocytosis," in which one cell "nibbles" another cell.SIGNIFICANCE STATEMENT The ingestion of the photoreceptor outer segment (OS) tips by the retinal pigment epithelium (RPE) is a dynamic cellular process that has fascinated scientists for 60 years. Yet its molecular mechanisms had not been addressed in living cells. We developed a live-cell imaging approach to investigate OS tip ingestion, and focused on the dynamic participation of actin filaments and membrane-shaping BAR proteins. We observed scission of OS tips for the first time, and were able to monitor local changes in protein concentration preceding, during, and following scission. Our approach revealed that actin filaments were concentrated at the site of OS scission and were required for regulating the size of the ingested OS tip and the time course of the ingestion process.

Inflammatory phenotype, clinicopathologic variables, and effects of long-term methylene blue in dogs with hereditary methemoglobinemia caused by cytochrome b5 reductase deficiency

To determine whether dogs with cytochrome b5 reductase (CYB5R) deficiency have a constitutive proinflammatory phenotype, characterize hematologic and serum chemistry results, and describe changes in methemoglobin (MetHb) levels and serum C-reactive protein (CRP) concentrations after long-term per os (PO) methylene blue (MB) therapy. 21 client-owned dogs (CYB5R deficient, n = 10; healthy controls, 11). In this prospective, case-control study, methemoglobin levels were measured using a blood gas analyzer with co-oximetry. Plasma tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10) concentrations were measured using a canine-specific multiplex bead-based assay. Serum CRP concentrations were measured with a canine-specific commercial ELISA kit. Serum CRP concentration and MetHb levels were measured in 6 dogs with CYB5R deficiency after ≥ 60 days of PO MB therapy. As expected, MetHb levels were higher in dogs with CYB5R deficiency compared to controls (P < .001). Plasma TNF-α, IL-6, IL-10, and serum CRP concentrations were no different between CYB5R-deficient and control dogs. Dogs with CYB5R deficiency had lower absolute lymphocyte (P = .005) and eosinophil counts (P = .04) and higher alanine transaminase (P = .04) and alkaline phosphatase activity (P = .02) than controls, but these changes were not clinically relevant. Methemoglobin levels decreased after PO MB therapy (P = .03). These results suggest that otherwise healthy dogs with CYB5R deficiency do not have a constitutive proinflammatory phenotype and clinically relevant abnormalities in hematologic and serum chemistry panels are not expected. Dogs with decreased quality of life attributed to methemoglobinemia from CYB5R deficiency might benefit from PO MB therapy.

High-Speed Dental Instruments: An Investigation of Protein-Contaminated Dental Handpieces with the Bicinchoninic Acid Assay in Dental Offices in Styria, Austria.

Due to permanent contact with bodily secretions such as blood and saliva, the dental workplace poses a high risk of infection for patients as well as for personnel. High-speed dental instruments are still considered one of the major hygienic risks, as the high-speed rotation of the attachments leads to the retraction of infectious material from patients' oral cavities. The aim of this study was to investigate the extent to which dental handpieces are contaminated after use. Spray-water samples were taken from different handpieces used in seven dental offices and protein concentrations were measured photometrically. In the first part of the study, samples were collected from each handpiece before and after the treatment of the patients. Additionally, the changes in protein concentration after consecutive treatments in which the same high-speed dental instrument was used were investigated. The results demonstrated measurable protein concentrations in 91.2% of a total of 398 samples, and 96.4% of the spray-water samples taken after treatment showed a discrepancy from the initial measured protein concentration. In 68.4% an increase in protein concentration was observed, whereas in 27.9% a decrease was measured. In conclusion, the internal contamination of high-speed dental instruments frequently occurs in daily usage and consequently may lead to the transmission of infectious agents by flushing the contaminated water out of the spray water tubes. Moreover, it must be pointed out that internal cleansing of handpieces is insufficient and that a final mechanical disinfection is indispensable.

Biomarkers of Drug Resistance in Temporal Lobe Epilepsy in Adults.

Temporal lobe epilepsy (TLE) is the most common type of focal epilepsy in adults. Experimental and clinical data indicate that neuroinflammation and neurodegeneration accompanying epileptogenesis make a significant contribution to the chronicity of epilepsy and the development of drug resistance in TLE cases. Changes in plasma and serum concentrations of proteins associated with neuroinflammation and neurodegeneration can be predictive biomarkers of the course of the disease. This study used an enzyme-linked immunosorbent assay of the following plasma proteins: brain-derived neurotrophic factor (BDNF), tumor necrosis factor alpha (TNFa), and high-mobility group protein B1 (HMGB1) in patients with mesial TLE to search for biomarkers of the disease. The objective of the study was to examine biomarkers of the neuroinflammation and neurodegeneration of plasma: BDNF, TNFa, and HMGB1. The aim of the study was to identify changes in the concentration of circulating pro-inflammatory and neurotrophic factors that are prognostically significant for the development of drug resistance and the course of TLE. A decrease in the concentration of BDNF, TNFa, and HMGB1 was registered in the group of patients with TLE compared with the control group. A significant decrease in the concentration of HMGB1 in patients with drug-resistant TLE was observed. Aberrations in plasma concentrations of BDNF, TNFa, and HMGB1 in patients with TLE compared with the controls have been confirmed by earlier studies. A decrease in the expression of the three biomarkers may be the result of neurodegenerative processes caused by the long course of the disease. The results of the study may indicate the acceptability of using HMGB1 and TNFa as prognostic biological markers to indicate the severity of the disease course and the risk of developing drug resistance.

Identifying the dataset to define the optimal timing of histopathological examination for central nervous system toxicity in MPTP-induced Parkinson's disease monkey model.

Determining the optimal timing for histopathological examination following exposure to a test article is crucial for assessing neurotoxicity. However, no study has focused on identifying an ideal dataset to define the optimal timing for histopathological examination of central nervous system (CNS) toxicity in monkeys. Therefore, this study aimed to define a predictive endpoint that would guide us in selecting the optimal timing for histopathological examination of CNS toxicity in monkeys. Four cynomolgus monkeys were administered 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intravenously at a dosage of 0.6 mg/kg twice at 1-week intervals. Necropsies were performed 1 week after the final dose. The Parkinsonian rating (PR) score and temporal changes in neurofilament light chain and glial fibrillary acidic protein concentrations in the cerebrospinal fluid (CSF) and serum were evaluated and compared with the histopathological findings in the brain. The PR score of all animals administered MPTP increased from days 10 to 11, with some degree of individual variability. Microscopically, all animals showed axonal swelling and vacuolation, with or without microgliosis in the nigrostriatal bundle. However, substantial neurodegenerative findings were observed only in animals with high PR scores at necropsy. A slight increase in CSF biomarker levels at necropsy was also observed in animals with high PR scores. However, their correlation with microscopic findings in these animals was unclear. These data suggest that comprehensive clinical observations, such as PR score alone or combined with other CSF biomarkers, could be further evaluated as potential indicators for triggering anatomic CNS evaluations in monkeys following toxic insults.

Water loss from silicone tubing and effect on protein concentration during drug product manufacturing

Silicone tubing is used in various unit operations during drug product (DP) manufacturing. Hold of protein formulations in silicone tubing over time may have an impact on product quality, particularly protein concentration. This study evaluated the change in protein concentration of a test monoclonal antibody (mAb) formulation over various hold times in silicone tubing as a function of tubing internal diameter (ID) and wall thickness. It was hypothesized that the rate of water diffusion through the semi-permeable membrane is a function of the tubing ID and wall thickness. The weight and protein concentration of various formulation-filled tubings over time was measured. The weight of water lost varied linearly with the change in protein concentration. It was observed to be independent of mAb type, formulation composition, and initial protein concentration for a given tubing ID and wall thickness. The effect of formulation water activity on the water loss rate was investigated. A mechanistic diffusion-based model was developed that predicts the change in tubing weight and therefore protein concentration over various hold times for a given formulation and tubing. Overall, this study suggests that water loss from silicone tubing affects protein concentration and should be monitored during DP process development and manufacturing.

Synapse-specific changes in Arc and BDNF in rat hippocampus following chronic temporal lobe epilepsy

Expression of immediate early genes (IEGs) in the brain is important for synaptic plasticity, and probably also in neurodegenerative conditions. To understand the cellular mechanisms of the underlying neuropathophysiological processes in epilepsy, we need to pinpoint changes in concentration of synaptic plasticity-related proteins at subsynaptic levels. In this study, we examined changes in synaptic expression of Activity-regulated cytoskeleton-associated (Arc) and Brai Derived Neurotrophic Factor (BDNF) in a rat model of kainate-induced temporal lobe epilepsy (TLE). Western blotting showed reduced concentrations of Arc and increased concentrations of BDNF in hippocampal synaptosomes in chronic TLE rats. Then, using quantitative electron microscopy, we found corresponding changes in subsynaptic regions in the hippocampus. Specifically, we detected significant reductions in the concentrations of Arc in the presynaptic terminal of Schaffer collateral glutamatergic synapses in the stratum radiatum of the CA1 area in TLE, as well as in their adjacent postsynaptic spines. In CA3, there was a significant reduction of Arc only in the presynaptic terminal cytoplasm. Conversely, in CA3, there was a significant increase in the expression of BDNF in the presynaptic terminal, but not in the postsynaptic spine. Significant increase in BDNF concentration in the CA1 postsynaptic density was also obtained. We hypothesize that the observed changes in Arc and BDNF may contribute to both cognitive impairment and increased excitotoxic vulnerability in chronic epilepsy.

Acute and chronic blood serum proteome changes in patients with methanol poisoning.

Twenty-four blood serum samples from patients with acute methanol poisoning (M) from the mass methanol poisoning outbreak in the Czech Republic in 2012 were compared with 46 patient samples taken four years after poisoning (S) (overlap of 10 people with group M) and with a control group (C) of 24 samples of patients with a similar proportion of chronic alcohol abuse. When comparing any two groups, tens to hundreds of proteins with a significant change in concentration were identified. Fifteen proteins showed significant changes when compared between any two groups. The group with acute methanol poisoning showed significant changes in protein concentrations for at least 64 proteins compared to the other groups. Among the most important identified proteins closely related to intoxication are mainly those involved in blood coagulation, metabolism of vitamin A (increased retinol-binding protein), immune response (e.g., increased complement factor I, complement factors C3 and C5), and lipid transport (increased apolipoprotein A I, apolipoprotein A II, adiponectin). For blood coagulation, the most affected proteins with significant changes in the methanol poisoning group were von Willebrand factor, carboxypeptidase N, alpha-2-antiplasmin (all increased), inter-alpha-trypsin inhibitor heavy chain H4, kininogen-1, plasma serine protease inhibitor, plasminogen (all decreased). However, heparin administration used for the methanol poisoning group could have interfered with some of the changes in their concentrations. Data are available via ProteomeXchange with the identifier PXD035726.

Skeletal muscle angiogenic, regulatory, and heat shock protein responses to prolonged passive hyperthermia of the human lower limb.

Passive hyperthermia induces a range of physiological responses including augmenting skeletal muscle mRNA expression. This experiment aimed to examine gene and protein responses to prolonged passive leg hyperthermia. Seven young participants underwent 3 h of resting unilateral leg heating (HEAT) followed by a further 3 h of rest, with the contralateral leg serving as an unheated control (CONT). Muscle biopsies were taken at baseline (0 h), and at 1.5, 3, 4, and 6 h in HEAT and 0 and 6 h in CONT to assess changes in selected mRNA expression via qRT-PCR, and HSP72 and VEGFα concentration via ELISA. Muscle temperature (Tm) increased in HEAT plateauing from 1.5 to 3 h (+3.5 ± 1.5°C from 34.2 ± 1.2°C baseline value; P < 0.001), returning to baseline at 6 h. No change occurred in CONT. Endothelial nitric oxide synthase (eNOS), Forkhead box O1 (FOXO-1), Hsp72, and VEGFα mRNA increased in HEAT (P < 0.05); however, post hoc analysis identified that only Hsp72 mRNA statistically increased (at 4 h vs. baseline). When peak change during HEAT was calculated angiopoietin 2 (ANGPT-2) decreased (-0.4 ± 0.2-fold), and C-C motif chemokine ligand 2 (CCL2) (+2.9 ± 1.6-fold), FOXO-1 (+6.2 ± 4.4-fold), Hsp27 (+2.9 ± 1.7-fold), Hsp72 (+8.5 ± 3.5-fold), Hsp90α (+4.6 ± 3.7-fold), and VEGFα (+5.9 ± 3.1-fold) increased from baseline (all P < 0.05). At 6 h Tm were not different between limbs (P = 0.582; CONT = 32.5 ± 1.6°C, HEAT = 34.3 ± 1.2°C), and only ANGPT-2 (P = 0.031; -1.3 ± 1.4-fold) and VEGFα (P = 0.030; 1.1 ± 1.2-fold) differed between HEAT and CONT. No change in VEGFα or HSP72 protein concentration were observed over time; however, peak change in VEGFα did increase (P < 0.05) in HEAT (+140 ± 184 pg·mL-1) versus CONT (+7 ± 86 pg·mL-1). Passive hyperthermia transiently augmented ANGPT-2, CCL2, eNOS, FOXO-1, Hsp27, Hsp72, Hsp90α and VEGFα mRNA, and VEGFα protein.

Genetic and Functional Effects of Adiponectin in Type 2 Diabetes Mellitus Development.

Diabetes mellitus (DM) is a common chronic metabolic disease, and the C57BLKsJ-db/db mice are good animal models for type 2 diabetes mellitus (T2DM). In this study, Western blotting and immunohistochemistry (IHC) were employed to examine the protein expression of adiponectin in the liver tissues of T2DM mice with different disease courses (4, 16, and 32 weeks). Adiponectin expression reduced in the liver tissues of T2DM mice in different disease courses. The genotypic and allelic frequencies of the adiponectin gene rs1063538 and rs2241766 single nucleotide polymorphisms (SNPs) in a Taiwanese population (570 T2DM patients and 1700 controls) were investigated. Based on the genetic distribution of the rs2241766 locus, the distribution frequency of the T allele in the T2DM group (72.8%) was higher than in the control group (68.8%). Individuals carrying the G allele had a 0.82-fold greater risk of developing T2DM than individuals carrying the T allele. Differences were evident in the genotypic and allelic distributions (p &lt; 0.05). Enzyme-linked immunosorbent assay (ELISA) was used to measure changes in serum adiponectin protein concentrations in the healthy population and in patients with T2DM. Serum adiponectin concentration in patients with T2DM was lower than in the control group. In summary, adiponectin was determined to be a T2DM susceptibility gene and may be involved in T2DM progression.

Perioperative Changes in Serum Concentration of Glial Fibrillar Acid Protein and Delayed Cognitive Recovery: an Experimental Observational Study

A number of studies have found an association between the increased concentration of glial fibrillar acid protein (GFAP) in blood serum in patients with various types of brain damage (ischemic stroke, traumatic brain injury, neurodegenerative and neuro-oncological diseases), as well as with a rapid decline in cognitive functions in elderly people with initially normal cognitive abilities.The objective: to identify the relationship between delayed cognitive recovery and changes in serum GFAP concentration in the perioperative period in patients operated for various oncological diseases.Subjects and Methods. The study included 30 patients who underwent surgical treatment for prostate cancer, colorectal cancer and pancreatic cancer under combined general anesthesia.The inclusion criteria were the expected duration of the operation over 300 minutes and the age over 60 years. GFAP was determined in plasma by enzyme immunoassay before anesthesia, the next day after surgery and on the 4th–5th day. Neuropsychological testing was performed before surgery and on the 4th–5th postoperative day. Delayed cognitive recovery was defined as a decrease in the composite z-score of more than one standard deviation (SD) compared to the preoperative assessment.Correlation analysis was performed between changes in the composite z-score (in absolute values) and the difference in GFAP concentration between the outcome and the first postoperative day, the outcome and the 4th–5th postoperative day and the first and 4th–5th postoperative days.Results. In five cases (16.6%), a decrease in the composite z-score &gt; 1 SD was revealed indicating a delayed cognitive recovery. In the remaining 25 (83.4%) patients, changes in the composite z-score were less than one standard deviation. The median concentration of GFAP in patients with delayed cognitive recovery was 0.13 [0.1; 0.14] before surgery, 0.12 [0.09; 0.14] the day after surgery and 0.16 [0.05; 0.19] on the 4th–5th day after surgery. In patients without cognitive impairment, the concentration of GFAP was 0.15 [0.125; 0.184] before surgery, 0.15 [0.121; 0.163] 24 hours after surgery and 0.13 [0.079; 0.151] on the 4th–5th day after surgery. The correlation values between changes in the composite z-score and the difference in GFAP concentrations were: between the outcome and the first postoperative day – rs = 0.107, p = 0.37, outcome and the 4th–5th postoperative day – rs = 0.134, p = 0.37, the first and 4th‒5th postoperative days – rs = 0.21, p = 0.37.Discussion. There was no statistically significant difference in GFAP levels between patients with delayed cognitive recovery and patients without cognitive impairment. There was also no correlation between the difference in GFAP concentrations in plasma before surgery and 24 hours after, before surgery and on the 4th–5th day of the postoperative period and the composite z-score.Conclusions. The use of GFAP to predict cognitive decline associated with surgical treatment of colorectal cancer, prostate cancer and pancreatic cancer under general anesthesia is not yet possible.

Microbial degradation of polystyrene microplastics by a novel isolated bacterium in aquatic ecosystem

The continuous accumulation of microplastics (MPs) in water, soil, and atmosphere will pose a serious threat to the environmental ecological balance. How to reduce and eliminate MPs in the environment has become a hot issue. Here we isolated Bacillus cereus CH6 from lake sediments and investigated its ability to degrade polystyrene microplastics(PS MPs). Bacillus cereus CH6 has short generation time, high activity, and fast substrate utilization. The weight loss was 10.7% by Bacillus cereus CH6 after 50 days of incubation. Moreover, the average degradation rate reached 0.967 mg/d. We further investigated the surface morphology changes and the internal structural changes during the degradation of PS MPs. Finally, the changes of bacterial protein concentration and esterase activity during the PS MPs degradation process were observed from the perspective of biological index changes. These results are expected to provide a new candidate for PS MPs degradation.

Trial of Antisense Oligonucleotide Tofersen for SOD1 ALS

BackgroundThe intrathecally administered antisense oligonucleotide tofersen reduces synthesis of the superoxide dismutase 1 (SOD1) protein and is being studied in patients with amyotrophic lateral sclerosis (ALS) associated with mutations in SOD1 (SOD1 ALS).MethodsIn this phase 3 trial, we randomly assigned adults with SOD1 ALS in a 2:1 ratio to receive eight doses of tofersen (100 mg) or placebo over a period of 24 weeks. The primary end point was the change from baseline to week 28 in the total score on the ALS Functional Rating Scale–Revised (ALSFRS-R; range, 0 to 48, with higher scores indicating better function) among participants predicted to have faster-progressing disease. Secondary end points included changes in the total concentration of SOD1 protein in cerebrospinal fluid (CSF), in the concentration of neurofilament light chains in plasma, in slow vital capacity, and in handheld dynamometry in 16 muscles. A combined analysis of the randomized component of the trial and its open-label extension at 52 weeks compared the results in participants who started tofersen at trial entry (early-start cohort) with those in participants who switched from placebo to the drug at week 28 (delayed-start cohort).ResultsA total of 72 participants received tofersen (39 predicted to have faster progression), and 36 received placebo (21 predicted to have faster progression). Tofersen led to greater reductions in concentrations of SOD1 in CSF and of neurofilament light chains in plasma than placebo. In the faster-progression subgroup (primary analysis), the change to week 28 in the ALSFRS-R score was −6.98 with tofersen and −8.14 with placebo (difference, 1.2 points; 95% confidence interval [CI], −3.2 to 5.5; P=0.97). Results for secondary clinical end points did not differ significantly between the two groups. A total of 95 participants (88%) entered the open-label extension. At 52 weeks, the change in the ALSFRS-R score was −6.0 in the early-start cohort and −9.5 in the delayed-start cohort (difference, 3.5 points; 95% CI, 0.4 to 6.7); non–multiplicity-adjusted differences favoring early-start tofersen were seen for other end points. Lumbar puncture–related adverse events were common. Neurologic serious adverse events occurred in 7% of tofersen recipients.ConclusionsIn persons with SOD1 ALS, tofersen reduced concentrations of SOD1 in CSF and of neurofilament light chains in plasma over 28 weeks but did not improve clinical end points and was associated with adverse events. The potential effects of earlier as compared with delayed initiation of tofersen are being further evaluated in the extension phase. (Funded by Biogen; VALOR and OLE ClinicalTrials.gov numbers, NCT02623699 and NCT03070119; EudraCT numbers, 2015-004098-33 and 2016-003225-41.)

PSVIII-21 Modulation of Goat Plasma Proteins Using Different Preparations of Garlic

Abstract The main objective of this study was to evaluate the effect of different garlic preparations on plasma protein concentration and Galectin secretion in goat blood. Inflammatory diseases caused by pathogens including gastrointestinal parasites impact goat gut health and productivity. Changes in plasma protein concentration are indicators of health status with application in inflammatory disease diagnosis and prognosis. Galectins (Gals) are small soluble proteins that contain one or two carbohydrate recognition domains (CRDS). ‘Prototype’ galectin-1, -2, -7, contain one CRD, Galectin 3 is chimeric, and galectin-4, -8, -9 and -12, are tandem repeat galectins with two CRDs. Galectins regulate biologic functions, immunity and disease pathogenesis and can serve as biomarkers. Garlic (Allium sativum) is a medicinal herb that has anti-parasitic and immuno-modulatory effects. Six (N=6) adult female clinically healthy Boer x Spanish goats were used. Blood was collected from the jugular vein and treated with different concentrations of fresh garlic, allicin, garlic barrier, or aged garlic (Sigma aldich). Total plasma protein secretion was evaluated using the BCA assay. Secretion of Gals 1(Prototype), Gal 3 (chimera type) and Gal 9 (tandem-repeat type) determined using commercial ELISA kits as recommended by the manufacturer. All variables were assessed using SAS analysis software (version 9.0, Sas Institute Cary, p≤0.05). Garlic treatment had a dose dependent effect on total plasma protein and Gals secretion. The protein secretion was affected by different types of garlic. Allicin (72.86μg/ml) reduced the concentration of plasma proteins. No change was observed with fresh garlic (156.334μg/ml) and garlic barrier (160.73μg/ml). Garlic barrier treatment increased protein secretion (6.3%) in goat blood. All Gals were detected (Gals 1 &amp;gt;9 &amp;gt;3), with a variable response to Garlic preparation (Garlic barrier &amp;gt; fresh garlic Gals secretion). Garlic modulates protein secretion. Studies are needed to determine the effect of different preparations on leukocytes and molecular targets in goat blood.

Dynamic changes in protein concentrations of keratins in crop milk and related gene expression in pigeon crops during different incubation and chick rearing stages

ABSTRACT 1. The objective of this study was to examine the keratin composition of crop milk, the variation of epithelial thickness and keratin (K) gene expression in samples from young pigeon during incubation and chick rearing. 2. Crop milk was collected from 1-, 3- and 5-day-old squab crops for keratin content analysis. Results showed that K4 accounted for the highest proportion of all detected keratins. 3. In total, 42 pairs of adult pigeons were allocated to seven groups according to different stages to collect crop samples. Gene expression studies showed that the K3 gene expression was maximised at rearing Day 15 (15) and R1 in males and females, respectively. K6a gene level was the greatest at R15 in females, whereas it peaked at incubation Day 4 (I4) in males. The K12, K13, K23 and K80 gene levels were inhibited at the peak period of crop milk formation in comparison with I4. In females, K cochleal expression peaked at I10, whereas it was the greatest at R25 in males. K4 and K14 gene expression was the highest at I10 in females, while K4 and K14 were minimised at I17 and R7 in males, respectively. Gene expressions of K5, K8, K19 and K20 in males and K19 in females were maximised at R1. The K5, K20 and K75 gene levels in females peaked at R7. K75 and K8 expressions in males and females reached a maximum value at R25 and I17, respectively. 4. The epithelial thickness of male and female crops reached their greatest levels at R1 and had the highest correlation with K19. 5. These results emphasised the importance of keratinisation in crop milk formation, and different keratins probably play various roles during this period. The K19 was probably a marker for pigeon crop epithelium development. The sex of the parent pigeon affected keratin gene expression profiles.

Increasing cell size remodels the proteome and promotes senescence

Cell size is tightly controlled in healthy tissues, but it is unclear how deviations in cell size affect cell physiology. To address this, we measured how the cell's proteome changes with increasing cell size. Size-dependent protein concentration changes are widespread and predicted by subcellular localization, size-dependent mRNA concentrations, and protein turnover. As proliferating cells grow larger, concentration changes typically associated with cellular senescence are increasingly pronounced, suggesting that large size may be a cause rather than just a consequence of cell senescence. Consistent with this hypothesis, larger cells are prone to replicative, DNA-damage-induced, and CDK4/6i-induced senescence. Size-dependent changes to the proteome, including those associated with senescence, are not observed when an increase in cell size is accompanied by an increase in ploidy. Together, our findings show how cell size could impact many aspects of cell physiology by remodeling the proteome and provide a rationale for cell size control and polyploidization.