Changes In Protein Metabolism Research Articles

Molecular analysis of human tick-bitten skin yields signatures associated with distinct spatial and temporal trajectories - A proof-of-concept study

Tick-associated diseases present challenges due to tridirectional interactions among host-specific responses, tick toxins and salivary proteins as well as microbes. We aimed to uncover molecular mechanisms in tick-bitten skin samples (cases) and contralateral skin samples (controls) collected simultaneously from the same participants, using spatial transcriptomics. Cases and controls analysed using NanoString GeoMx Digital Spatial Profiler identified 274 upregulated and 840 downregulated differentially expressed genes (DEGs), revealing perturbations in keratinization and immune system regulation. Samples of skin biopsies taken within 72 h post tick-bite DEGs had changes in protein metabolism and viral infection pathways as compared to samples taken 3 months post tick-bite, which instead displayed significant perturbations in several epigenetic regulatory pathways, highlighting the temporal nature of the host response following tick bites. Within-individual signatures distinguished tick-bitten samples from controls and identified between-individual signatures, offering promise for future biomarker discovery to guide prognosis and therapy.

Acute effects of testosterone on whole body protein metabolism in hypogonadal and eugonadal conditions: a randomized, placebo-controlled, crossover study.

Chronic testosterone (T) substitution and short-term T administration positively affect protein metabolism, however, data on acute effects in humans are sparse. This study aimed to investigate T's acute effects on whole body protein metabolism in hypogonadal and eugonadal conditions. We designed a randomized, double-blind, placebo-controlled, crossover study, including 12 healthy young males. Whole body protein metabolism was evaluated during 1) eugonadism, and after medically induced hypogonadism, with application of a gel on each trial day containing either 2) placebo, 3) T 50 mg, or 4) T 150 mg; under basal (5-h basal period) and insulin-stimulated conditions (3-h clamp). The main outcome measure was a change in net protein balance. The net protein loss was 62% larger in the placebo-treated hypogonadal state compared with the eugonadal state during the basal period (-5.5 ± 3.5 µmol/kg/h vs. -3.4 ± 1.2 µmol/kg/h, P = 0.038), but not during the clamp (P = 0.06). Also, hypogonadism resulted in a 25% increase in whole body urea flux (P = 0.006). However, T did not result in any significant changes in protein breakdown, synthesis, or net balance during either the basal period or clamp (all P > 0.05). Protein breakdown was reduced during clamp compared with the basal period regardless of gonadal status or T exposure (all P ≤ 0.001). In conclusion, the application of transdermal T did not counteract the negative effects of hypogonadism with no effects on protein metabolism within 5 h of administration. Insulin (during clamp) mitigated the effects of hypogonadism. This study is the first to investigate acute protein metabolic effects of T in hypogonadal men.NEW & NOTEWORTHY In a model of medically induced hypogonadism in male volunteers, we found increased whole body urea flux and net protein loss as an expected consequence of hypogonadism. Our study demonstrates the novel finding that the application of transdermal testosterone had no acute effects on whole body protein metabolism under eugonadal conditions, nor could it mitigate the hypogonadism-induced changes in protein metabolism. In contrast, insulin (during clamp) mitigated the effects of hypogonadism on protein metabolism.

The role of acute phase inflammatory proteins in the pathogenesis of Metabolic syndrome in obese horses

Metabolic syndrome and obesity in horses are closely related processes that are accompanied by chronic inflammation. Our research aimed to establish the normative indicators of acute phase inflammation proteins in obese horses. We conducted a study on twenty horses, ten of which were in the control group and ten were in the experimental group, showing signs of obesity. The Henneke scoring system was used to assess the body condition. This system ranks animals from one to nine, with one being exhausted and nine being very fat. Serum concentrations of circulating immune complexes, seromucoids, C-reactive protein, haptoglobin, total protein, and its fractions were determined. It was found that significant changes in protein metabolism occur in animals with obesity, namely: the content of circulating immune complexes, seromucoids, C-reactive protein, haptoglobin, and globulins significantly increase, indicating the development of inflammatory processes in horses due to obesity. The localization of these processes is associated both with obesity in animals and possibly with laminitis, the development of which is one of the pathogenetic links of metabolic syndrome

Current Concepts of Molecular Mechanisms of Age-Related Changes in Eye Lens

Age-related cataract is the leading cause of vision loss in old people worldwide. According to the World Health Organization it accounts for 47.8 % of the total number of ocular pathologies in people over 50 years old. Despite the rapid development of cataract surgery technology, surgery remains a challenge due to its cost and the increasing number of patients. Literature review is devoted to current concepts of pathogenesis and molecular mechanisms of age-related changes in eye lens. There are the three main theories of cataractogenesis: oxidative stress; the impact of quinoid substances, which are formed due to the impairment of aromatic amino acid metabolism and the activation of aldo reductase enzyme with subsequent accumulation of sorbitol, reactive oxygen species (ROS) generation, dysfunction of Na+/K+ channels and calcium deregulation causing lens epithelial cells apoptosis. Theories of pathogenesis are linked and based on the development of age-related changes in protein metabolism (the majority of nuclear α-crystallins are insoluble), glucose metabolism (non-enzymatic glycosylation of proteins), lipid metabolism, enzyme activity and the loss of membrane potential of cells ( the increase of Na+ and Ca2+ level and the decrease of K+ level). Key element of all theories of age-related changes in eye lens is the aggregation of high molecular weight proteins covalent-bonded of disulfide linkages. Based on molecular mechanisms of cataractogenesis, the development of pathogenetically oriented medical methods of correction of the age-related changes in lens is carried on. This review provides information on results of experimental and clinical studies which demonstrate the anti-cataract effect of Pirenoxine 0, 005 %.

Protein Metabolism Changes and Alterations in Behavior of Trace Amine-Associated Receptor 1 Knockout Mice Fed a High-Fructose Diet

Trace amines and their receptors are a family of G protein-coupled receptors widely distributed in the central nervous system and periphery. The trace amine-associated receptor 1 (TAAR1) plays a significant role as a therapeutic target for schizophrenia, depression, diabetes, and obesity. In this study, TAAR1 knockout mice and WT groups were tested in conditions of a high-fructose diet. The consumption of a high-fructose diet may be due to the influence on the metabolism processes by dopamine in the brain, neuromotor function, and level of anxiety of TAAR1 knockout mice. During a comparative analysis of behavioral, biochemical, and morphological parameters, significant differences were found between liver and biochemical parameters, the regulation of protein metabolism (AST/ALT ratio, creatine kinase activity, urea), and alterations in behavior. An elevated plus maze analysis showed the influence of fructose and genetic factors on the level of anxiety. A new marker of the grooming microstructure (depression ratio) was tested, which showed high efficiency as a marker of depression-like behavioral changes and a possible association with dopamine-dependent regulation of protein metabolism. These results confirm a possible association of the TAAR1 gene knockout with an increase in catabolic reaction levels by AST/ALT-dependent and possible dopamine-mediated protein metabolism regulation and depression-like behavior.

Proteomic Analysis of Roots Response to Potassium Deficiency and the Effect of TaHAK1-4A on K+ Uptake in Wheat.

Potassium (K+) is essential for plant growth and stress responses. A deficiency in soil K+ contents can result in decreased wheat quality and productivity. Thus, clarifying the molecular mechanism underlying wheat responses to low-K+ (LK) stress is critical. In this study, a tandem mass tag (TMT)-based quantitative proteomic analysis was performed to investigate the differentially abundant proteins (DAPs) in roots of the LK-tolerant wheat cultivar "KN9204" at the seedling stage after exposure to LK stress. A total of 104 DAPs were identified in the LK-treated roots. The DAPs related to carbohydrate and energy metabolism, transport, stress responses and defense, and post-translational modifications under LK conditions were highlighted. We identified a high-affinity potassium transporter (TaHAK1-4A) that was significantly up-regulated after the LK treatment. Additionally, TaHAK1-4A was mainly expressed in roots, and the encoded protein was localized in the plasma membrane. The complementation assay in yeast suggested that TaHAK1-4A mediates K+ uptake under extreme LK conditions. The overexpression of TaHAK1-4A increased the fresh weight and root length of Arabidopsis under LK conditions and improved the growth of Arabidopsis athak5 mutant seedlings, which grow poorly under LK conditions. Moreover, silencing of TaHAK1-4A in wheat roots treated with LK stress decreased the root length, dry weight, K+ concentration, and K+ influx. Accordingly, TaHAK1-4A is important for the uptake of K+ by roots exposed to LK stress. Our results reveal the protein metabolic changes in wheat induced by LK stress. Furthermore, we identified a candidate gene potentially relevant for developing wheat lines with increased K+ use efficiency.

N-3 Fatty Acid Supplementation Impacts Protein Metabolism Faster Than it Lowers Proinflammatory Cytokines in Advanced Breast Cancer Patients: Natural 15N/14N Variations during a Clinical Trial.

While clinical evidence remains limited, an extensive amount of research suggests a beneficial role of n-3 polyunsaturated fatty acid supplementation in cancer treatment. One potential benefit is an improvement of protein homeostasis, but how protein metabolism depends on proinflammatory cytokines in this context remains unclear. Here, using the natural abundance of the stable isotopes of nitrogen as a marker of changes in protein metabolism during a randomized, double-blind, controlled clinical trial, we show that protein homeostasis is affected way faster than proinflammatory cytokines in metastatic breast cancer patients supplemented with n-3 polyunsaturated fatty acids. We provide some evidence that this response is unrelated to major changes in whole-body substrate oxidation. In addition, we demonstrate that more fatty acids were impacted by metabolic regulations than by differences in their intake levels during the supplementation. This study documents that the percentage of patients that complied with the supplementation decreased with time, making compliance assessment crucial for the kinetic analysis of the metabolic and inflammatory responses. Our results highlight the time-dependent nature of metabolic and inflammatory changes during long-chain n-3 fatty acid supplementation.

Migratory disposition alters lean mass dynamics and protein metabolism in migratory white-throated sparrows (Zonotrichia albicollis).

Migratory birds seasonally increase fat stores and the capacity to use fat to fuel long-distance migratory flights. However, lean mass loss also occurs during migratory flights and, if adaptive, should exhibit seasonal changes in the capacity for protein metabolism. We conducted a photoperiod manipulation using captive white-throated sparrows (Zonotrichia albicollis) to investigate seasonal changes in protein metabolism between the nonmigratory "winter" condition and after exposure to a long-day "spring" photoperiod to stimulate the migratory condition. After photostimulation, birds in the migratory condition rapidly increased fat mass and activity of fat catabolism enzymes. Meanwhile, total lean mass did not change, but birds increased the activity of protein catabolism enzymes and lost more water and lean mass during water-restricted metabolic testing. These data suggest that more protein may be catabolized during migratory seasons, corresponding with more water loss. Counter to predictions, birds in the migratory condition also showed an approximately 30-fold increase in muscle expression of sarcolipin, which binds to sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) and uncouples Ca2+ transport from ATP hydrolysis. Our documented changes to protein catabolism enzymes and whole animal lean mass dynamics may indicate that protein breakdown or increased protein turnover is adaptive during migration in songbirds.

Effect of thymogen immunocorrected boar castration on biochemical indicators

The article presents the results of the dynamics of indicators of protein, nitrogen and carbohydrate metabolism during castration of boars in industrial production against the background of immunocorrection with thymogen. The studies were carried out on 14 boars (aged 5 months), selected by random sampling from 38 animals in compliance with the principle of paired analogues and divided into two groups ‒ control and experimental ones (n=7 in each). All boars were castrated by an open method with preoperative medical support according to the methodology adopted in the farm. Animals of the experimental group were additionally injected with thymogen for a five-day course. During the experiment, the content of C-reactive and total proteins, albumin, globulin, urea, creatinine, glucose was determined in the blood serum. Albumin-globulin ratio was determined by the calculation method. According to the results of the experiment, it was found that castration of boars was accompanied in both groups at all stages of the experiment by an increase in the content of C-reactive protein, changes in protein metabolism, mainly in the catabolic phase of the postoperative period ‒ a decrease in the concentration of total protein, globulin below the norm in the control group with similar dynamics of indicators in the experimental group, but within the reference intervals. Further dynamics were positive in the direction of normalization at the anabolic phase. The dynamics of urea and creatinine in both groups had a wave-like character, not going beyond the normative values, the level of urea increased with a subsequent decrease, creatinine – vice versa. Their final levels were lower than the original ones.The use of thymogen largely leveled the intensity of changes in the catabolic phase of the pathological process and contributed to the normalization of the biochemical profile at the final stage of the experiment.

Features of changes in protein metabolism in patients with polytrauma depending on the timing of surgical treatment of injuries of the musculoskeletal system

The evaluation of the peculiarities of protein metabolism changes depending on the terms of surgical treatment of the loco-motor system injuries was petformed in patients with polytrauma. It was shown that plasma fibrinogen increase, posttraumatic metabolic azotemia, i.e. increase of blood urea and creatinine levels, decrease of total protein level were present in all patients. In the same time disproteinemia, i.e. decrease of albumin levels and increase of globulin fractions (more significantly alpha-1 and alpha-2 globulins) was noted. In contrast to the delayed treatment early surgical intervention was accompanied by redistribution of the fraction composition of proteins in a form of the earlier normalization of the relative quantity of alpha-1 and alpha-2 globulins. This is an evidence of the plastic exchange restoration and possibilities of compensatory responses of the organism.

Search for a way to improve the serum albumin binding function and the functional state of the liver when hypertension combined with non-alcoholic fatty liver disease

Recently, there has been a significant increase in interest in research on hypertension (HT), primarily due to its high prevalence. The interest in studying this problem is also exacerbated by the often insufficient effectiveness of existing treatments. The effect of concomitant pathologies on HT, in particular non-alcoholic fatty liver disease (NAFLD), remains poorly understood. The aim of the study – to evaluate the changes in the serum albumin binding function (SABF) and its relationship with the biochemical parameters of the blood when HT and HT combined with NAFLD and to suggest ways of medical correction of the detected disorders. Material and methods. 76 individuals with stage 2 HT with degree 2–3 arterial hypertension were examined. They were divided into two groups. Group 1 included 28 patients with HT without concomitant diseases who received basic hypertension therapy, and group 2 included patients with concomitant NAFLD. The latter in turn was divided into two subgroups: 2a – 27 patients who in addition to basic HT therapy received additional Antral hepatoprotector 200 mg three times a day for 2 months, and 2b – 21 patients who received only basic HT therapy. All of them underwent a standard clinical examination, as well as SABF, protein fractions, and liver function indicators. The comparison group consisted of 25 healthy individuals, comparable in age and sex. Results and Discussion. Patients in group 1 showed moderate changes in the functional state of the liver, but they did not exceed the norm, patients in group 2 – a significant decrease in SABF, as well as protein metabolism (decrease in total protein, albumin, albumin-globulin ratio and increase globulins) and liver function (increased activity of aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltranspeptidase, thymol levels, alkaline phosphatase and total bilirubin). After treatment, the majority of patients in subgroup 2 had a statistically significant increase in SABF and a quantitative improvement in protein fractions and functional state of the liver. In subgroup 2-b, where hepatoprotective treatment was not performed, significant changes in most indicators did not occur. The results may be related to the positive effect of the drug on the liver, which leads to improved functional status of hepatocytes and their protein-synthesizing ability. In subgroup 2 b, where hepatoprotective treatment was not performed, significant changes in most indicators did not occur. The results may be related to the positive effect of the drug on the liver leading to improved functional status of hepatocytes and their protein-synthesizing ability. Conclusions. Changes of the functional state of the liver are observed when HT without concomitant pathology. HT with NAFLD is accompanied by a significant decrease in SABF, changes in protein metabolism and the functional state of the liver. Prescribing Antral to such patients helps to increase SABF, normalize protein metabolism and improve the functional state of the liver.

THE EFFECT OF PLACENTAL CRYOEXTRACT ON THE STATE OF PROTEIN-LIPID METABOLISM IN THE GASTRIC MUCOSA IN EXPERIMENTAL STRESS-INDUCED ULCERS

Introduction. Peptic ulcer is one of the most prevalent diseases of the gastrointestinal tract. Stress factor is considered to have the highest impact on the development of ulcers, as it is present in almost all cases of onset and exacerbation of this disease. Acute stress ulcers occur with severe injuries, acute diseases of various organs, shock, a sharp drop in blood pressure, oxygen deficiency of body tissues, liver, kidney and others. Given the above facts, in the correction of stress-induced lesions of the gastric mucosa (GM) therapeutically, the target should consider not only reducing the aggression factors of gastric juice, but also the normalization of changes in protein and carbohydrate metabolism in GM. In this aspect, our attention was drawn to the domestic biotechnological preparation of placental cryoextract (CEP), which has a range of valuable biological effects. The aim is to study the effect of cryopreserved placenta extract on the state of protein-lipid metabolism in the gastric mucosa in a model of water-immobilization stress in rats. Materials and methods of research. The studies were performed on 28 nonlinear laboratory male rats weighing 200–220 g. Stress-induced gastric ulcer was modeled under water-immobilization stress (WIS) in rats according to the method of Takagi et al. To obtain the homogenate, the GM was perfused with cold (+ 4°C) buffer solution and homogenized at 3000 rpm (teflon/glass). The content of oxidatively modificated proteins (OMP) in GM was determined by Dubinina spectrophotometric method. The content of total lipids in the GM was determined spectrophotometrically by color reaction with sulfophosphovaniline reagent. Phospholipids (PL) were fractionated by the method of Svetashev and Vaskovsky. Research results. Evaluation of changes in protein metabolism in GM showed that the level of total protein in rats, which were preventively administered CEP before WIS, was almost comparable with that of intact rats, i.e., 50.1 ± 1.7 μg/mg tissue and 51.1 ± 1.3 μg/mg of tissue, respectively, which indicated the elimination of disorders of protein homeostasis with the introduction of the studied cryoextract. Studies of changes in total lipids and PL showed that the content of PL of animals treated with CEP was 26.9 ± 0.9%, which was not significantly different from that of intact animals (30.5 ± 0.9%) and, at the same time, it was by 7.3% higher (p < 0.001) as compared with animals who were administered esomeprazole. Conclusions. Prophylactic five-day administration of CEP to WIS leads to normalization of all evaluated indices, in particular, to increase of the total protein level by 29.0% (p <0.01), decrease of oxidatively modified proteins level by 20.6% (p <0.01), and the 2.3-fold increase (p <0.001) of the level of phospholipids in the total lipids pool.

Ovarian cancer ascites induces skeletal muscle wasting in vitro and reflects sarcopenia in patients.

BackgroundCachexia‐associated skeletal muscle wasting or ‘sarcopenia’ is highly prevalent in ovarian cancer and contributes to poor outcome. Drivers of cachexia‐associated sarcopenia in ovarian cancer remain elusive, underscoring the need for novel and better models to identify tumour factors inducing sarcopenia. We aimed to assess whether factors present in ascites of sarcopenic vs. non‐sarcopenic ovarian cancer patients differentially affect protein metabolism in skeletal muscle cells and to determine if these effects are correlated to cachexia‐related patient characteristics.MethodsFifteen patients with an ovarian mass and ascites underwent extensive physical screening focusing on cachexia‐related parameters. Based on computed tomography‐based body composition imaging, six cancer patients were classified as sarcopenic and six were not; three patients with a benign condition served as an additional non‐sarcopenic control group. Ascites was collected, and concentrations of cachexia‐associated factors were assessed by enzyme‐linked immunosorbent assay. Subsequently, ascites was used for in vitro exposure of C2C12 myotubes followed by measurements of protein synthesis and breakdown by radioactive isotope tracing, qPCR‐based analysis of atrophy‐related gene expression, and NF‐κB activity reporter assays.ResultsC2C12 protein synthesis was lower after exposure to ascites from sarcopenic patients (sarcopenia 3.1 ± 0.1 nmol/h/mg protein vs. non‐sarcopenia 5.5 ± 0.2 nmol/h/mg protein, P < 0.01), and protein breakdown rates tended to be higher (sarcopenia 31.2 ± 5.2% vs. non‐sarcopenia 20.9 ± 1.9%, P = 0.08). Ascites did not affect MuRF1, Atrogin‐1, or REDD1 expression of C2C12 myotubes, but NF‐κB activity was specifically increased in cells exposed to ascites from sarcopenic patients (sarcopenia 2.2 ± 0.4‐fold compared with control vs. non‐sarcopenia 1.2 ± 0.2‐fold compared with control, P = 0.01). Protein synthesis and breakdown correlated with NF‐κB activity (r s = −0.60, P = 0.03 and r s = 0.67, P = 0.01, respectively). The skeletal muscle index of the ascites donors was also correlated to both in vitro protein synthesis (r s = 0.70, P = 0.005) and protein breakdown rates (r s = −0.57, P = 0.04).ConclusionsAscites of sarcopenic ovarian cancer patients induces pronounced skeletal muscle protein metabolism changes in C2C12 cells that correlate with clinical muscle measures of the patient and that are characteristic of cachexia. The use of ascites offers a new experimental tool to study the impact of both tumour‐derived and systemic factors in various cachexia model systems, enabling identification of novel drivers of tissue wasting in ovarian cancer.

INFLUENCE OF CHRONIC NON-GASEOUS ACIDOSIS ON BONE TISSUE

The urgency of the problem is due to the high prevalence among the population in all countries of the world and socio-economic losses associated with serious complications of osteoporosis. Chronic non-gaseous acidosis leads to disruption of many metabolic processes, which causes dysfunction of some organs, namely, can lead to decreased kidney function, exacerbate cardiovascular disease, the development of osteoporosis. Different types of non-gaseous acidosis are accompanied by changes in protein metabolism, which lead to the development of osteoporosis.&#x0D; Aim: to study the structural features of the bone tissue of the femur in chronic non-gaseous acidosis.&#x0D; Methods. The experiments were carried out on 60 intact white outbred male rats weighing 120-180 g, which were kept on a balanced diet of vivarium. Chronic non-gaseous acidosis was simulated by daily intragastric administration with a probe for 60 days 20 mmol/kg NH4CL. The control animals were injected with the same amount of tap water. Histological examination and osteometry were used to study the structure, length and thickness of the rat femur. Free, bound, and total oxyproline in rat urine was determined by biochemical method.&#x0D; Results. In chronic non-gaseous acidosis after 6 months there is a tendency to reduce the length and thickness of the femur, but does not differ significantly from the control group. In the urine of rats of the experimental group, the total oxyproline increases mainly due to an increase in free oxyproline, which indicates the predominance of collagen degradation over repair. The histological structure of the bone is disturbed due to the thinning of the bone beams, the intercellular matrix is disorganized, in some places there is its "defibering" with the formation of foci of destruction and cylindrical "lumens". There is a violation of the crystalline structure of bone tissue, its uneven calcification, the formation of cracks in the bone beams.&#x0D; Studied literature sources and our research indicate that chronic non-gaseous acidosis has a pathogenic effect on the histological structure of bone. Acidosis is a stress factor that increases the level of glucocorticoids. Glucocorticoids inhibit type I collagen synthesis and increase the expression of collagenase 3, which promotes collagen degradation. Collagen is a source of amino acids that are the substrate for renal ammoniogenesis. Renal ammoniogenesis is a compensatory mechanism in chronic non-gaseous acidosis, which required for restores the normal ratio between fixed cations and anions blood plasma. Thus, the violation of the organic basis of bones is the body's adaptation to chronic non-gaseous acidosis.&#x0D; Conclusions. Chronic non-gaseous acidosis simultaneously leads to thinning and disorganization of the intercellular matrix, disruption of the crystal structure of bone tissue, which indicates its important role in the development of osteoporosis. In chronic non-gaseous acidosis, the concentration of oxyproline in the urine increases, which is a marker of the destruction of collagen in the organic matrix of bone.

Impact of β−hydroxy-β−methylbutyrate (HMB) on muscle loss and protein metabolism in critically ill patients: A RCT

Muscle wasting deteriorates life quality after critical illness and increases mortality. Wasting starts upon admission to intensive care unit (ICU). We aimed to determine whether β-hydroxy-β-methylbutyrate (HMB), a metabolite of leucine, can attenuate this process. Prospective randomized, placebo-controlled double blind trial. ICU patients depending on mechanical ventilation on day 3 having a functional gastrointestinal tract. They were randomized to HMB (3g/day) or placebo (maltodextrin) from day 4 on for 30 days. magnitude of loss of skeletal muscle area (SMA) of the quadriceps femoris measured by ultrasound at days 4 and 15. body composition, change in protein metabolism assessed by amino acids tracer pulse, and global health at 60 days. Data are mean [95% CI]. Statistics by ANCOVA with correction for confounders sex, age and/or BMI. Thirty patients completed the trial, aged 65 [59, 71] years, SAPS2 score 48 [43, 52] and SOFA 8.5 [7.4, 9.7]. The loss of total SMA was 11% between days 4 and 15 (p<0.001), but not different between the groups (p=0.86). In the HMB group, net protein breakdown (Δ Estimate HMB-Placebo:-153 [-242,-63]; p=0.0021) and production of several amino acid was significantly reduced, while phase angle increased more (0.66 [0.09, 1.24]; p=0.0247), and SF-12 global health improved more (Δ Estimate HMB-Placebo: 27.39 [1.594, 53.19], p=0.04). HMB treatment did not significantly reduce muscle wasting over 10 days of observation (primary endpoint), but resulted in significantly improved amino acid metabolism, reduced net protein breakdown, a higher phase angle and better global health. CLINICALTRIALS. NCT03628365.

Impact of phenolic compounds on free amino acid contents and electrophoretic patterns of the Nile Tilapia, Oreochromis niloticus

The present research was designed to determine the free amino acids (FAAs) composition and electrophoretic protein patterns by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in Oreochromis niloticus exposed to 1/10 LC50-96 hrs of phenol and nonylphenol ethoxylate 9 (NPEO 9) under laboratory condition for 4 weeks, in an attempt to develop sensitive bio-indicators for evaluation of pollution effects. Variations of FAAs were apparent in both treated groups. Significant increases were recorded in all individual amino acids including essential and non-essential ones, except for proline for both treated groups and leucine for phenol exposed fish group where they decreased significantly. The electrophoretic analysis of protein of the skeletal muscle homogenate using SDS-PAGE indicated that the control group yielded 10 fractions of molecular masses between 166 and 23 kDa. A total number of 9 bands were separated from fish of phenol group; they were varied from 168 to 23 kDa. However, 6 bands only were separated from fish of the NPEO 9 group; they were varied from 157 to 28 kDa. The muscle protein extract from fish of the phenol group showed five unique bands (168, 96, 68, 52, and 39 kDa). Moreover, fish that were exposed to NPEO 9 group showed three unique bands (157, 92, and 72 kDa). The highest similarity index S value observed between phenol and control groups was 0.37, followed by 0.33 between NPEO 9 group and the control one. This could be an indicator of changes in protein metabolism due to phenol and NPEO 9 pollution.

LC-MS/MS analysis of lesional and normally looking psoriatic skin reveals significant changes in protein metabolism and RNA processing.

BackgroundPlaque psoriasis is a chronic autoimmune disorder characterized by the development of red scaly plaques. To date psoriasis lesional skin transcriptome has been extensively studied, whereas only few proteomic studies of psoriatic skin are available.AimThe aim of this study was to compare protein expression patterns of lesional and normally looking skin of psoriasis patients with skin of the healthy volunteers, reveal differentially expressed proteins and identify changes in cell metabolism caused by the disease.MethodsSkin samples of normally looking and lesional skin donated by psoriasis patients (n = 5) and samples of healthy skin donated by volunteers (n = 5) were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After protein identification and data processing, the set of differentially expressed proteins was subjected to protein ontology analysis to characterize changes in biological processes, cell components and molecular functions in the patients’ skin compared to skin of the healthy volunteers. The expression of selected differentially expressed proteins was validated by ELISA and immunohistochemistry.ResultsThe performed analysis identified 405 and 59 differentially expressed proteins in lesional and normally looking psoriatic skin compared to healthy control. In normally looking skin of the patients, we discovered decreased expression of KNG1, APOE, HRG, THBS1 and PLG. Presumably, these changes were needed to protect the epidermis from spontaneous activation of kallikrein-kinin system and delay the following development of inflammatory response. In lesional skin, we identified several large groups of proteins with coordinated expression. Mainly, these proteins were involved in different aspects of protein and RNA metabolism, namely ATP synthesis and consumption; intracellular trafficking of membrane-bound vesicles, pre-RNA processing, translation, chaperoning and degradation in proteasomes/immunoproteasomes.ConclusionOur findings explain the molecular basis of metabolic changes caused by disease in skin lesions, such as faster cell turnover and higher metabolic rate. They also indicate on downregulation of kallikrein-kinin system in normally looking skin of the patients that would be needed to delay exacerbation of the disease. Data are available via ProteomeXchange with identifier PXD021673.

Вплив цистицеркозної інвазії на показники протеїнового обміну та клітинний імунітет кролів

The composition of blood proteins depends on the functional state of animals, as well as on various pathologies. The goal of the work was to establish the effect of cysticercosis invasion on the protein metabolism and cellular immunity of rabbits. Rabbits were divided into two groups after visual identification of the larval cysts presence: healthy (control) and diseased (experimental). Biochemical blood tests were performed using reagent sets produced by “Filisit-Diagnostika” (Ukraine). Spontaneous cysticercosis invasion of male rabbits caused changes in protein metabolism. The content of total protein in infected animals was higher by 8.79% (P&lt;0.05) than in control. The increase in the total protein content was due to the globulin fraction, which was 1.50 times (P&lt;0.05) higher in affected animals than in healthy rabbits. This rise occurred as a result of a high content of γ-globulins, both in absolute value by 1.69 times (P&lt;0.05), and in percentage by 1.51 times (P&lt;0.05). The albumins, which are completely synthesized by liver, were lower by 10.08% (P&lt;0.05) in rabbits with cysticercosis. The protein coefficient of experimental animals was lower by 43.89% (P&lt;0.05) compared with the control. The concentration of uric acid in diseased rabbits was lower by 34.09% (P&lt;0.05) than in healthy ones. We have observed a high number of T-, B-lymphocytes and T-helper cells, especially the last two indicators, which increased respectively 1.54 times (P&lt;0.01) and 1.36 times (P&lt;0.05) against the background of a low number of T-suppressors and O-lymphocytes — almost 5 times (P&lt;0.001) in affected animals than in healthy rabbits. In our opinion, such changes in the proteinogram and cellular immunity indicate the intensification of immune reactions in the body of rabbits infected with the Cysticercus pisiformis as a result of inflammatory processes that occur in the liver under the action of a mechanical, toxic effect of the helminths.

Hematological parameters and protein metabolism in the blood of pregnant rats under the effect of vanadium citrate

Dose-dependent changes in protein metabolism in the blood and hematological parameters of pregnant rats under the effect of vanadium citrate are presented in the article. The animals were divided into five groups: group I – non-pregnant females, II – pregnant females consuming pure water without additives, III, IV, V – females which during the mating and pregnancy period received the solution of vanadium citrate at concentrations of 0.03, 0.125 and 0.50 μg V/mL water. The research findings show that in pregnant animals of group II, the level of urea and alkaline phosphatase activity increased, meanwhile aspartate aminotransferase activity decreased, as compared to the non-pregnant females of group І. The levels of total protein and albumin decreased; however, the content of β-globulins increased in the pregnant animals of group II, as compared with that in group I. Also, in the rats of group II, there was a decrease in hemolysis time, total content of erythrocytes and hemoglobin, the content of old and mature erythrocytes, while the content of young erythrocytes increased, as compared to group I. The platelet content and thrombocrit in rats of group II increased in comparison with group I. The content of leukocytes and lymphocytes in pregnant animals of group II decreased, while the content of granulocytes increased, in contrast to non-pregnant rats. Under the effect of vanadium citrate at concentrations of 0.03–0.50 μg V/mL, there was a significant increase in the maximum number of prohemolized erythrocytes, the time of maximum hemolysis was delayed by 0.4–0.6 min, as compared with the pregnant rats of group II. This did not affect the time of total hemolysis in rats of groups III and V, as compared with the pregnant animals in group II. Under the effect of vanadium citrate, an increase in the content of young erythrocytes was observed, as compared with group II. The hemoglobin content decreased at the concentration of 0.125 μg V/mL, while at the concentration of 0.50 μg V/mL it increased, as compared to the pregnant animals of group II. Also, under the effect of vanadium citrate there was a decrease in the mean hemoglobin concentration in the erythrocyte. In pregnant animals fed with vanadium citrate solutions, the platelet content and thrombocrit, the relative width of platelet distribution by volume decreased, as compared with the pregnant rats of group II. The content of leukocytes, lymphocytes and granulocytes under the effect of vanadium citrate increased, as compared with the pregnant animals in group II. Under the effect of vanadium citrate at the concentration of 0.03 μg V/mL, the level of albumin, creatinine and aspartate aminotransferase activity increased in blood plasma in comparison with group II. Meanwhile, at the concentration of 0.125 μg V/mL, the relative content of γ-globulins and aspartate aminotransferase activity increased, alkaline phosphatase activity and urea level decreased in comparison with group II. However at the concentration of 0.50 μg V/mL, the relative α- and γ-globulins content and aspartate aminotransferase activity increased, at the same time, the relative β-globulins content and urea level decreased in comparison with group II. Therefore, vanadium citrate normalizes the indicators of protein metabolism during pregnancy, thus it can be considered as a potential dietary drug for the pregnant.

Investigation of the level of endogenous intoxication of the body and the characteristics of the saliva crystallization in poultry with feed toxicosis

In the course of the experiments, it was found that during the reproduction of feed toxicosis in broiler chickens caused by prolonged intake of mycotoxins (T-2 toxin and aflatoxin B1), clinical signs of intoxication were recorded starting from the 5th day of the experiment. Disturbances in the biochemical profile of the bird’s blood were manifested by changes in protein metabolism, as well as in indicators reflecting the functioning of the liver and kidneys. The level of endogenous intoxication in the body of broiler chickens with combined mycotoxicosis was significantly higher than the data of intact control at a spectrophotometer wavelength λ = 254 nm - by 14.8%, at λ = 280 nm - by 36.6%, AWM distribution index (DI) 280/254 increased by 18.9%. Comparison of the saliva facies of healthy chickens and chickens with mycotoxicosis revealed differences between the studied crystallograms.