Tissue Protein Content Research Articles

Dual Effects of Beta-Hydroxy-Beta-Methylbutyrate (HMB) on Amino Acid, Energy, and Protein Metabolism in the Liver and Muscles of Rats with Streptozotocin-Induced Type 1 Diabetes.

Beta-hydroxy-beta-methyl butyrate (HMB) is a unique product of leucine catabolism with positive effects on protein balance. We have examined the effects of HMB (200 mg/kg/day via osmotic pump for 7 days) on rats with diabetes induced by streptozotocin (STZ, 100 mg/kg intraperitoneally). STZ induced severe diabetes associated with muscle wasting, decreased ATP in the liver, and increased α-ketoglutarate in muscles. In plasma, liver, and muscles increased branched-chain amino acids (BCAAs; valine, isoleucine, and leucine) and decreased serine. The decreases in mass and protein content of muscles and increases in BCAA concentration were more pronounced in extensor digitorum longus (fast-twitch muscle) than in soleus muscle (slow-twitch muscle). HMB infusion to STZ-treated animals increased glycemia and serine in the liver, decreased BCAAs in plasma and muscles, and decreased ATP in the liver and muscles. The effects of HMB on the weight and protein content of tissues were nonsignificant. We concluded that fast-twitch muscles are more sensitive to STZ than slow-twitch muscles and that HMB administration to STZ-treated rats has dual effects. Adjustments of BCAA concentrations in plasma and muscles and serine in the liver can be considered beneficial, whereas the increased glycemia and decreased ATP concentrations in the liver and muscles are detrimental.

C1QTNF6 Overexpression Acts as a Predictor of Poor Prognosis in Bladder Cancer Patients.

Background Bladder cancer is one of the most common urinary malignancies. This study is aimed at providing some promising molecular biomarkers for bladder cancer (BC) by investigating the correlation between C1QTNF6 expression and clinical characteristics as well as prognosis in patients with bladder cancer. Methods Sequencing profiles of C1QTNF6 mRNA in BC patients were collected to evaluate the distinctive gene expression, between normal bladder mucosa and BC, according to the TCGA and GEO databases. The association between C1QTNF6 expression and the clinical features as well as the disease prognosis was evaluated using two independent cohorts. The expression of C1QTNF6 in normal bladder and BC cells was examined by western blotting and PCR, so the underlying molecular mechanism could be further investigated. Results C1QTNF6 mRNA levels were found to be differentially expressed in two independent public cohorts, including the TCGA database and GSE13507 dataset from GEO. The protein and RNA levels of C1QTNF6 in BC cells were both elevated when compared to normal bladder cell lines. High C1QTNF6 expression was detected in advanced T/M stages, pathological grade, and AJCC stage when compared to the low C1QTNF6 expression group. The underlying mechanism related to this differential expression could be explained by cell migration and invasion assays, where bladder cancer cells 5637 and T24 had a significant reduction on migration and invasion ability upon knockdown of C1QTNF6 expression. The low C1QTNF6 expression group presented a more prominent OS advantage over the high-expression group in both TCGA and GSE13507 cohorts. Moreover, the protein content in tissues was further validated using the HPA database and TMA. Survival analyses also indicated that the high C1QTNF6 expression group had an unfavorable OS when compared to the low-expression group. Conclusions High C1QTNF6 expression may serve as a predictor of poor prognosis in bladder cancer patients, and the underlying mechanism is possibly associated with changes on cancer cell migration and invasion ability.

Management of gummosis in citrus with potassium phosphite

This work aimed to evaluate the potassium phosphite-based commercial product, Phytogard®, as a complementary tool for the management of gummosis in citrus. Seedlings of tangerine ‘Sunki’ were sprayed at concentrations 0; 0.5; 2 and 5 mL L-1 of Phytogard® and subsequently inoculated with zoospores of Phytophthora nicotianae. The disease incidence was reduced by 84% in plants sprayed at the concentration 0.5 mL L-1 and the plants sprayed at concentrations of 2 and 5 mL L-1 showed zero disease incidence. There was increased an production of fresh matter of shoot and roots in plants sprayed and inoculated with the pathogen compared to unsprayed plants. Plants sprayed with Phytogard® and inoculated with the pathogen showed lower values for total phenols, enzyme activity for phenylalanine ammonia-lyase and peroxidase and for total protein content in root tissues compared to non-inoculated plants. There was higher activity of the enzyme β 1.3-glucanase in root tissues of plants inoculated with the pathogen that received the product at the concentration of 2 mL L-1. The results of this study showed that the potassium phosphite-based product Phytogard® has potential for the control of Phytophthora nicotianae in seedlings of tangerine ‘Sunki’. However, it is not possible to conclude that this control occurs through resistance induction.

Influence of graded level of dietary protein with equated level of limiting amino acids on growth, feed utilization, body indices and nutritive profile of snubnose pompano, Trachinotus blochii (Lacepede, 1801) reared in low saline water

An experiment was conducted to delineate the effect of varying level of dietary protein with balanced level of limiting amino acids such as lysine and methionine on growth, feed utilization, body indices and digestive enzymology of snubnose pompano, Trachinotus blochii reared in low saline water. There were four experimental diets formulated with 350 (CP350), 400 (CP400), 450 (CP450) and 500 g/kg (CP500) crude protein and each with 100 g/kg crude fat and were assigned to four treatments, CP350, CP400, CP450 and CP500 respectively. Totally eighty four fingerlings with the body weight in the range of 16−18 g were randomly distributed in four treatment groups with each of three replicates. Fish were fed to apparent satiation twice daily with the assigned experimental diet. At the end of 13 weeks of feeding trial, weight gain percentage (WG%) and specific growth rate (SGR) of fish fed with CP400 and CP450 diets were significantly higher than those of fish fed with CP350 and CP500 diets. Growth rate on metabolic body weight (GRMBW), daily growth coefficient (DGC) and average daily growth (ADG) were higher in CP400 group and lower in CP350 group. The viscerosomatic index (VSI), hepatosomatic index (HSI), gastrosomatic index (GSI) and intra-peritoneal fat ratio (IPFR) showed an increasing trend as the crude protein content of the experimental diet increases and the maximum value was witnessed in CP500 treatment. Crude lipid and protein content of whole body tissue was found highest in CP400 and lowest level was noticed in CP350 group. The maximum intestinal protease activity was noticed in CP400 group. In conclusion, analysis of second degree polynomial regression revealed that the optimum dietary protein level could be in the range of 395.3–426.7 g/kg for maximum growth of juvenile snubnose pompano.

Spatial heterogeneity and temporal dynamics of protein-degrading activity and life-history traits in threespine stickleback Gasterosteus aculeatus

The paper studies the spatial heterogeneity and temporal dynamics of life-history traits, parasite invasion, and biochemical variables (e.g., tissue protein content and calpain activity) in adult and juvenile threespine stickleback Gasterosteus aculeatus. Samples were collected on three spawning grounds in Kandalaksha Bay, on the White Sea, located within a few kilometers of each other. These sites featured differences in temperature regime, water exchange, feed organism composition, predation load, aquatic vegetation, and spawner density. Whereas we found no parasite-related or size heterogeneity in adult fish on these grounds at the beginning of the spawning season, muscle protein reserves and protease (calpain) activity patterns in adults showed significant spatial heterogeneity, which increased as spawning progressed and was particularly pronounced in females. Sticklebacks expend much energy during spawning and are worn out by the end of the spawning period. To maintain individual viability, skeletal muscle proteins degrade, and the resulting amino acids are oxidized to produce energy. Protein-degrading calpains play a key role in these processes, and also mediate intense protein metabolism in juvenile stickleback, showing higher values in individuals growing under more favorable conditions. Thus, by studying the heterogeneous life-history traits and biochemical characteristics of adult and juvenile stickleback, we discovered the effects of environmental factors on their physiology, biochemical variables, and growth.

Seasonal effects of heavy metals on the date mussel Lithophaga lithophaga (Mollusca:Bivalvia) at Eastern harbor, Alexandria, Egypt

The edible mussel Lithophaga lithophaga is considered as one of the most important human food sources in Alexandria, Mediterranean Sea. The present study is designed to determine the seasonal bioaccumulation levels of Cd, Co and Pb in the whole soft tissues as well as different tissues of Lithophaga lithophaga. Results revealed that the seasonal bioaccumulation levels of Cd, Co and Pb in date mussel were below the permissible limits or other reported values from other regions of the Mediterranean. On the other hand, the order of metals accumulation level in different tissues was as follows: digestive gland>remaining soft tissues>gonads. The present study confirmed the role of digestive gland as a concentration center for heavy metals. Moreover, total protein content and stress protein responses of the whole soft tissues were evaluated. The total protein content was arranged in the following order: summer>autumn>spring>winter. Five novel stress proteins appeared in summer. The histological and ultrastructural studies of the digestive gland of Lithophaga lithophaga collected in summer and spring showed marked histopathological alternations.

Protein metabolism, physicochemical properties and chemical composition of muscle tissue in Large White weaners

We presented the blood serum biochemical parameters, physicochemical properties and chemical composition of muscle tissue in Large White weaners of British origin. We found that serum biochemical parameters in five months weaners were within the physiological norm (the content of total protein was 71.28 g/l (?V = 8.16%), creatinine concentration was 155.62 µmol L-1 (?V = 14.13%). Studies of physicochemical properties and chemical composition of samples of Longissinus dorsi muscle of Large White young pigs show that the number of high quality samples in terms of moisture holding capacity, % is 12.0%, colour intensity, units of ext.×1000 – 16.0%, tenderness, s – 12.0% and fat content, % – 16.0%. We revealed significant correlations between moisture holding capacity and protein content (r = -0.484, tr = 2.65), moisture holding capacity and hygroscopic moisture content (r = 0.402, tr = 2.11). The correlation between total protein content in blood serum and fat content in muscle tissue was -0.257 (tr = 1.28), whereas the correlation between serum total protein content and calcium content in muscle tissue was 0.375 (tr = 1.94).

Albinism in plants - far beyond the loss of chlorophyll: Structural and physiological aspects of wild-type and albino royal poinciana (Delonix regia) seedlings.

The partial or complete loss of chlorophylls, or albinism, is a rare phenomenon in plants. In the present study, we provide the first report of the occurrence in albino Delonix regia seedlings and describe the morpho-physiological changes associated with albinism. Wild-type (WT) and albino seedlings were characterized. Leaflets samples were processed following common procedures for analysis with light, scanning and transmission electron microscopy. The chlorophyll a fluorescence parameters and the carbohydrate, lipid and soluble protein content were also determined in leaf and cotyledon samples of both albino and WT seedlings. Albino seedlings showed reduced growth. They also had lower chlorophyll and protein content in foliar tissues than WT seedlings, in addition to lower concentrations of lipids and carbohydrates stored in cotyledons. The chloroplasts of albino seedlings were poorly developed, with an undefined internal membrane system and the presence of plastoglobules. Wild-type seedlings had a uniseriate and hypoestomatic epidermis. The mesophyll was dorsiventral, consisting of a layer of palisade parenchyma and two to four layers of spongy parenchyma. In albino seedlings, the spongy parenchyma was compact, with few intercellular spaces, and the thickness of the mesophyll was larger, resulting in increased thickness of the leaf blade. Albino seedlings had higher stomatal density and number of pavement cells, although the stomata had smaller dimensions. In addition to the partial loss of chlorophylls, albino D.regia showed changes at physiological and structural levels, demonstrating the crucial nature of photosynthetic pigments during the development and differentiation of plant leaf tissues/cells.

Development of technology for canning of ringed seal meat

Results of studies on technology of natural canned meat of ringed seal (akiba) are presented. The frozen seal meat was used for canning without subcutaneous fat that was removed after slaughter. Protein content in the muscle tissue was 22.50 ± 0.71 %, fat content — 8.20 ± 0.20 %, minerals — 1.20 ± 0.10 %, its energy value was 167.4 ± 5.6 kcal/100 g. After defrosting, pieces of the seal meat were kept in fresh water at temperature of 15–18 oC for 2 to 3 hours, then onion and flavorings were added. The spiced meet was packed to cans № 6 and sterilized under temperature of 120 oC. The finished canned food had a meaty taste and smell, without extraneous smacks and smells. The broth was transparent with brown tint and small amount of suspended protein substances. Protein content in the finished canned food was 17.0 %, fat content 7.0 %, protein composition was close to the standard protein by amount and ratio of essential amino acids. Monounsaturated fatty acids dominated in lipids of the canned food (58.08 % of total lipids), mainly oleic (18:1 n-9) and palmitooleic (16:1 n-7) acids; portion of polyunsaturated fatty acids was 22.17 %, including omega-3 fatty acids (18.56 % of PUFA). High content of iron (6.6 mg/100 g), zinc (2.3 mg/100 g) and selenium (80 μg/100 g) was detected in the canned food that partially or completely maintained daily requirements of human body for these elements.

Effects of High-Intensity Exercise Training on Adipose Tissue Mass, Glucose Uptake and Protein Content in Pre- and Post-menopausal Women.

The menopausal transition is accompanied by changes in adipose tissue storage, leading to an android body composition associated with increased risk of type 2 diabetes and cardiovascular disease in post-menopausal women. Estrogens probably affect local adipose tissue depots differently. We investigated how menopausal status and exercise training influence adipose tissue mass, adipose tissue insulin sensitivity and adipose tissue proteins associated with lipogenesis/lipolysis and mitochondrial function. Healthy, normal-weight pre- (n = 21) and post-menopausal (n = 20) women participated in high-intensity exercise training three times per week for 12 weeks. Adipose tissue distribution was determined by dual-energy x-ray absorptiometry and magnetic resonance imaging. Adipose tissue glucose uptake was assessed by positron emission tomography/computed tomography (PET/CT) by the glucose analog [18F]fluorodeoxyglucose ([18F]FDG) during continuous insulin infusion (40 mU·m−2·min−1). Protein content associated with insulin signaling, lipogenesis/lipolysis, and mitochondrial function were determined by western blotting in abdominal and femoral white adipose tissue biopsies. The mean age difference between the pre- and the post-menopausal women was 4.5 years. Exercise training reduced subcutaneous (~4%) and visceral (~6%) adipose tissue masses similarly in pre- and post-menopausal women. Insulin-stimulated glucose uptake, assessed by [18F]FDG-uptake during PET/CT, was similar in pre- and post-menopausal women in abdominal, gluteal, and femoral adipose tissue depots, despite skeletal muscle insulin resistance in post- compared to pre-menopausal women in the same cohort. Insulin-stimulated glucose uptake in adipose tissue depots was not changed after 3 months of high-intensity exercise training, but insulin sensitivity was higher in visceral compared to subcutaneous adipose tissue depots (~139%). Post-menopausal women exhibited increased hexokinase and adipose triglyceride lipase content in subcutaneous abdominal adipose tissue. Physical activity in the early post-menopausal years reduces abdominal obesity, but insulin sensitivity of adipose tissue seems unaffected by both menopausal status and physical activity.

ВОЗДЕЙСТВИЕ ПРИРОДНОГО ГАЗА НА АКТИВНОСТЬ ТКАНИ СЕЛЕЗЕНКИ В ОНТОГЕНЕЗЕ

The chronobiological approach and biochemical research methods have revealed the age-related features of the ultradian rhythms of the content of total protein, albumin in rat spleen tissue normal, under the influence of natural gas and against the background of the introduction of a complex of antioxidants (vitamin E and thymalin). During the experiment, the rhythmic activity of the total protein and albumin content in rat spleen tissue during ontogenesis was compared with chronic exposure to sulfur-containing natural gas and the introduction of an antioxidant complex against intoxication. The revealed fluctuations in the content of total protein and albumin can be characterized as ultradian rhythms with periods from 20-40 to 60 minutes. The obtained experimental data indicate a decrease in the stability of spleen tissue with age and depletion of the antioxidant system of rats with aging. The materials of the article are of practical value for the development of methods for correcting the negative effects of oxidative stress in the context of intoxication.

Effects of a Ketogenic Diet Containing Medium-Chain Triglycerides and Endurance Training on Metabolic Enzyme Adaptations in Rat Skeletal Muscle

Long-term intake of a ketogenic diet enhances utilization of ketone bodies, a particularly energy-efficient substrate, during exercise. However, physiological adaptation to an extremely low-carbohydrate diet has been shown to upregulate pyruvate dehydrogenase kinase 4 (PDK4, a negative regulator of glycolytic flux) content in skeletal muscle, resulting in impaired high-intensity exercise capacity. This study aimed to examine the effects of a long-term ketogenic diet containing medium-chain triglycerides (MCTs) on endurance training-induced adaptations in ketolytic and glycolytic enzymes of rat skeletal muscle. Male Sprague-Dawley rats were placed on either a standard diet (CON), a long-chain triglyceride-containing ketogenic diet (LKD), or an MCT-containing ketogenic diet (MKD). Half the rats in each group performed a 2-h swimming exercise, 5 days a week, for 8 weeks. Endurance training significantly increased 3-oxoacid CoA transferase (OXCT, a ketolytic enzyme) protein content in epitrochlearis muscle tissue, and MKD intake additively enhanced endurance training–induced increases in OXCT protein content. LKD consumption substantially increased muscle PDK4 protein level. However, such PDK4 increases were not observed in the MKD-fed rats. In conclusion, long-term intake of ketogenic diets containing MCTs may additively enhance endurance training–induced increases in ketolytic capacity in skeletal muscle without exerting inhibitory effects on carbohydrate metabolism.

Hepatic toxicological responses of SiO2 nanoparticle on Oreochromis mossambicus

The present study was carried out to investigate the impact of various concentrations of SiO2 nanoparticles (SiO2 NP) on the commonly available freshwater fish Oreochromis mossambicus. The 96 h median lethal concentration (LC50) of SiO2 NP was found to be between 270−280 ppm. This novel study has demonstrated histological alterations in the hepatic tissues and a dose-dependent depletion of tissue protein content and an elevated transaminases activity in the treated fish, which has facilitated understanding of the impact of SiO2 NP in O. mossambicus.

Sublethal concentrations of triclosan elicited oxidative stress, DNA damage, and histological alterations in the liver and brain of adult zebrafish.

Triclosan (TCS), an antimicrobial agent, has been a pollutant of increasing concern owing to its potential health risk on humans and aquatic animals. The present study seeks to test the hypothesis that TCS could alter the oxidative stress-related parameters in the brain and liver, as well as eliciting DNA damage in hepatocytes of adult zebrafish. On the basis of the 96h LC50 (398.9μg/L), adult zebrafish were separately exposed to 50, 100, and 150μg/L TCS for 30days. The brain and liver tissues from adult zebrafish were excised and assayed for a suite of antioxidant parameters and oxidative stress biomarkers including DNA damage in the liver. The induced effect by TCS on the activity of acetylcholinesterase (AChE) was also analyzed in the brain. Results showed a significant decrease in superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in the brain and liver of adult zebrafish. Also, the contents of the glutathione system (GSH and GSSH), as well as the activity of the glutathione reductase (GR), assayed in the liver, were reduced while the contents of malondialdehyde (MDA) were elevated in the liver. A comet assay revealed dose-dependent DNA damage in zebrafish hepatocytes. The 8-hydroxy-2'-deoxyguanosine (8-OHdG), MDA, and carbonyl protein contents in brain tissues significantly increased. Moreover, the AChE in the zebrafish brain was induced. Apparently, no obvious histological changes in brain tissues of zebrafish were observed compared with those of the control whereas atrophy and necrosis of hepatocytes and increased hepatic plate gap were observed in zebrafish hepatocytes after TCS exposure. The obtained results highlight that sublethal concentrations of TCS may be deleterious to the liver and brain of adult zebrafish upon subchronic exposure.

Tissue-specific bioaccumulation, metabolism and excretion of tris (2-ethylhexyl) phosphate (TEHP) in rare minnow (Gobiocyprisrarus)

Tris (2-ethylhexyl) phosphate (TEHP) is one of the most commonly used organophosphorus flame retardant (OPFR) analogues and is commonly detected in surface water and sediments. Limited information is available about the metabolic pathway or metabolite formation related to TEHP in fish. In this study, rare minnows (Gobiocyprisrarus) were exposed to TEHP in static water for 30 d to investigate the bioaccumulation and metabolite distribution in the fish muscle, liver, kidney, gill, GI-tract, ovary and testis. Based on the estimated kup,parent and kd,parent values, the bioconcentration factors (BCFparent) of TEHP in fish tissues were calculated in the order of kidney > ovary ≈ liver ≈ testis > gill ≈ GI-tract > muscle; this finding was consistent with the results of our previous study on other alkyl-substituted OPFRs. In addition, this study identified the metabolic profiles of TEHP in the liver. TEHP was oxidatively metabolized by the fish to a dealkylated metabolite (di 2-ethylhexyl phosphate; DEHP) and hydroxylated TEHP (OH-TEHP). OH-TEHP further underwent extensive phase II metabolism to yield glucuronic acid conjugates. DEHP was mainly distributed in rare minnow in the following order: liver > GI-tract > kidney ≫ other tissues. However, the metabolite showed lower accumulation potential in fish tissues than TEHP, with metabolite parent concentration factors (MPCFs) for DEHP of less than 0.1 in all the investigated tissues. The BCFparent values of TEHP in various fish tissues were only 9.0 × 10−3-7.2 × 10−4 times its estimated tissue-water partition coefficient (Ktissue-water) values based on tissue lipid, protein and water contents, which indicated the significance of biotransformation in reducing the bioaccumulation potential of TEHP in fish. The toxicokinetic data in the present study help in understanding the tissue-specific bioaccumulation and metabolism pathways of TEHP in fish and highlight the importance of toxicology research on TEHP metabolites in aquatic organisms.

Bradford assay as a high-throughput bioanalytical screening method for conforming pathophysiological state of the animal

Proteins are the essential components of the tissues that play a key role in the body. Its expression in the cell or tissue under a specified set of conditions and at a particular time regulates the different body conditions either as a normal body function or as a disease state. Protein is an important building block of muscles, skin, cartilage, bones and blood. Bradford assay is a reliable advanced and cost-effective protein estimation test for determining the exact concentration of protein in different tissues of the animal. In this study, we have taken a rat suffering from protein deficiency disorder and total protein concentration in the heart, brain, liver, blood and kidney was determined. It was found that the total protein concentration in different tissues of rat i.e., heart, brain, liver, plasma and kidney was found to be 8.39 ± 0.75, 10.46 ± 0.76, 6.74 ± 0.39, 8.12 ± 0.32 mg/g of tissue and 61.27 ± 0.95 mg/mL of plasma respectively (mean ± SEM). As compared to earlier published reports the total protein concentration in different tissues like hear, brain, liver and kidney found much lower to standard value as reported by Beyer, the reason behind obtaining this kind of results may be due to the presence of insufficient amount of the protein content in different tissue of animal as suffering from protein degeneration disorder. The rat was unable to digest and store the protein or catabolism was much faster than
 anabolism.
 Keywords: Anabolism, Bradford assay, Catabolism, Protein estimation.

Compensatory growth of Nile tilapia Oreochromis niloticus , L. subjected to cyclic periods of feed restriction and feeding in a biofloc system

The compensatory growth, productive performance, proximal composition and somatic indices of Nile tilapia (Oreochromis niloticus) cultivated in biofloc were evaluated during a 144-day period under five cyclic regimes of feed restriction and feeding. Five treatment groups, in which the frequency of feed restriction (R) and feeding (F) varied by periods (days) as follows: R1:F3, R3:F9, R6:F18, R8:F24 and R12:F36; each treatment was evaluated in triplicate. The cycles were repeated throughout the culture period. The control group received feed daily. Fish were cultivated in 18 circular tanks (3 m3) at a density of 50 fish/m3 per tank. At the end of the study, the survival of Nile tilapia was greater than 90% in all the treatments. Complete compensation in growth was achieved in R6:F18 and R12:F36. At the end of the feed restriction period, both crude protein and total lipid content of the tilapia muscle tissue taken from fish of the treatment groups were similar to samples of muscle tissue derived from fish of the control group; however, a reduction of more than 40% in somatic indices compared with the control was observed, but these recovered by the end of the feeding phase. The results indicate that cyclic feeding based on 12 days of feed restriction and 36 days of feeding (R12:F36) induced a complete compensation in weight and restoration of energy reserves, with similar measures of productive performance observed when compared to the control treatment during the culture of Nile tilapia in biofloc, and food reduction did not affect proximal composition.

P641 An increased baseline mucosal TNF burden linked to adalimumab non-response: opportunities for therapeutic drug monitoring

Abstract Background Various studies report robust associations between serum adalimumab (ADM) exposure and therapeutic outcome, which has led to the implementation of reactive therapeutic drug monitoring. As very little is known how serum levels relate to tissue and faecal ADM levels and to tumour necrosis factor (TNF) levels, we measured serum, tissue and faecal drug and TNF, and questioned how all this is linked to the therapeutic efficacy of ADM. Methods We prospectively recruited paired serum, mucosal biopsies and stools from 15 patients with ulcerative colitis (UC, n = 8) and Crohn’s disease (CD, n = 7) initiating ADM. All received ADM standard dosage. Samples were collected at baseline and week 8 (UC) or month 6 (CD). Endoscopic remission was defined as a Mayo endoscopic sub-score 0 (UC) or complete absence of ulcerations (CD). ADM levels were measured using a homogeneous immunoassay in which the ADM-TNFα complex is formed and captured. TNFα and IL-6 were quantified using an in-house developed immunoassay. Total tissue protein content was quantified in order to normalise the data. Results Pre-treatment tissue TNF and IL6 levels correlated significantly (r = 0.8, p < 0.001) and were lower in ADM remitters (n = 8) in contrast to non-remitters (4.5 vs. 8.6 pg/mg, p = 0.03; 7.1 vs. 122.5 pg/mg, p = 0.05). Pre-treatment faecal TNF was numerically lower in remitters (0.4 vs. 1.0 pg/mg, p = 0.30), but did not correlate with tissue/serum TNF (p = 0.84, p = 0.58). Although ADM did not significantly impact post-exposure serum or faecal TNF and IL6, it did reduce tissue TNF and IL6 (p = 0.02, p = 0.003), both in remitters (median −38.3%, p = 0.04; median −86.0%, p = 0.04) and non-remitters (median −59.5%, p = 0.16; median −84.5%, p = 0.03). Although TNF and IL6 changes did not correlate with ADM exposure or change in calprotectin, the decrease in faecal calprotectin significantly correlated with tissue, but not serum, ADM levels (r = 0.74, p = 0.01). ADM exposure was significantly higher in remitters as compared with non-remitters, both in serum (14.6 vs. 5.9 mg/ml, p = 0.009) and in tissue (53.1 vs. 24.1 ng/mg, p = 0.04). However, serum and tissue ADM levels correlated only moderately (r = 0.5, p = 0.1). Finally, tissue TNF/tissue ADM ratios were significantly higher in non-remitters, both considering pre (p = 0.007) and post TNF (p = 0.03). Faecal loss of ADM could be documented in a single patient only. Conclusion Both in tissue and serum ADM exposure was significantly higher in remitters, though tissue and serum ADM exposure did not perfectly correlate. A higher baseline tissue TNF burden was associated with non-response to ADM, and may help in the differentiation to select the appropriate biological in a given patient: no anti-TNF or intensified dosing in patients with a high baseline tissue TNF burden.

THE EFFECT OF ACUTE HYPEGLYCEMIA ON THE ACTIVITY OF THE ANTIOXIDANT SYSTEM IN THE CENTRAL NERVOUS SYSTEM AND IN THE LIVER OF THE INVERTEBRATE MOLLUSK - LYMNAEA STAGNALIS

Comparison of dynamic changes of biochemical reactions in the liver and CNS of freshwater liver mollusc LYMNAEA STAGNALIS has been conducted. It has been conducted during incubation in the highly concentrated solution of glucose. The indicators of superoxiddismutase (SOD) activity, the level of renewed glutation (G-SH), Se-dependant glutation peroxidase (Se-GP) and TBA-active products were estimated and compared. Also for the tissues of the Central Neural System and tissues samples of liver it was found out by glucoseoxidate method the concentration of glucose in haemolymph of the investigated biological object in the conditions of hyperglycemia. A comparative estimation of the general protein concentration in both tissues was conducted. Resistance of the liver tissue to hyperglycemia and a reverse effect towards CNS was found out.

PH Gradient Mitigation in the Leaf Cell Secretory Pathway Attenuates the Defense Response of Nicotiana benthamiana to Agroinfiltration.

Partial neutralization of the Golgi lumen pH by the ectopic expression of influenza virus M2 proton channel is useful to stabilize acid-labile recombinant proteins in plant cells, but the impact of pH gradient mitigation on host cellular functions has not been investigated. Here, we assessed the unintended effects of M2 expression on the leaf proteome of Nicotiana benthamiana infiltrated with the bacterial gene vector Agrobacterium tumefaciens. An isobaric tags for relative and absolute quantification quantitative proteomics procedure was followed to compare the leaf proteomes of plants agroinfiltrated with either an "empty" vector or an M2-encoding vector. Leaves infiltrated with the empty vector had a low soluble protein content compared to noninfiltrated control leaves, associated with increased levels of stress-related proteins but decreased levels of photosynthesis-associated proteins. M2 expression partly compromised these effects of agroinfiltration to restore soluble protein content in the leaf tissue, associated with restored levels of photosynthesis-associated proteins and reduced levels of stress-related proteins in the apoplast. These data illustrate the cell-wide influence of the Golgi lumen pH homeostasis on the leaf proteome of N. benthamiana responding to microbial challenge. They also underline the relevance of assessing the eventual unintended effects of accessory proteins used to modulate specific cellular or metabolic functions in plant protein biofactories.