Biomarkers Of Energy Metabolism Research Articles

Analysis of the season-dependent component in the evaluation of morphological and biochemical blood parameters in Shetland ponies of both sexes during exercise.

Determination of morphological and biochemical blood indices facilitates assessment of the health and welfare of horses, their nutrient demand, the effects of training already undertaken, and the horses' suitability for exercise. Identification of the season-dependent components and the effects of sex and exercise on changes in frequently referenced haematological and biochemical parameters was the main goal of the current study. The blood morphology of 21 healthy adult Shetland ponies (11 mares and 10 stallions) aged 6.5 ± 1.4 years from the central Pomeranian region in Poland was analysed. Blood samples were taken once per season for one year. No statistically significant season-dependent differences were found in the blood morphology parameters in either mares or stallions before or after exercise. Beta-coefficient results revealed the strength and type of the relationship of red blood cell distribution width (RDW) and granulocyte count (GRA) with the season, of red blood cell count (RBC), haematocrit, mean corpuscular volume and mean platelet volume with the sex, and of RDW, white blood cell count, GRA and RBC with the exercise factor. Biomarkers demonstrating the relationship between aerobic and anaerobic levels of energy metabolism in the blood did not show any sex dependency in regression analysis. The sex-independence of energy metabolism biomarkers may indicate the universality of these parameters. Both seasonality itself and its combination with the exercise factor took part in the formation of effective adaptive reactions for maintenance of morphological blood indices in the ponies during exercise.

Abstract 102: Imaging Markers Of Energy Metabolism In The Post-stroke Mouse Brain

Introduction: Cerebral ischemia results in a lesion comprised of dying cells surrounded by a band of tissue containing cells at risk of dying, called the penumbra. Understanding the status of energy metabolism in the stroke is necessary to get a clearer picture of the formation of the lesion, and may provide insights into potential therapeutic targets. The detection and distribution of energy metabolism biomarkers, particularly in ischemic brain tissue, presents significant challenges for conventional laboratory techniques due to the labile nature of these markers and the lack of unique chemical or spectroscopic handles. Fourier Transform infrared (FTIR) spectroscopic imaging can be used to map the distribution of different biochemical parameters, such as lipids, proteins, aggregated proteins, glycogen, lactate, pyruvate, ATP/ADP, NADH and glutamate. Methods: We employed the photothrombotic stroke model in adult mice to produce a permanent focal stroke lesion. Brain tissue was collected at multiple time points (1h to 4 weeks post-stroke). FTIR imaging was employed to map the distribution of unique biochemical fingerprints for over a dozen key metabolic markers. Immunohistochemistry was also performed on adjacent tissues to correlate biomarkers revealed by FTIR with the distribution of astrocytes, myelin and macrophages. Results: During the first week poststroke, the lesion shows decreased lipid esters, protein, ATP/ADP, NADH content while showing an increase in glutamate, aggregated protein, lactate and pyruvate. Glycogen can be seen to accumulate in the penumbra starting at 1 day post-stroke until 1 week post-stroke, which correlates with the appearance of astrocytes around the border of the stroke lesion. From 1 week to 4 weeks post-stroke, we observe an increase in lipid esters in the lesion, as well as a decrease in glycogen, lactate and pyruvate, while an increase in ATP/ADP and NADH can be observed. Conclusion: We are showing for the first time novel images of multiple energy metabolism biomarkers across a range of post-stroke time points in a mammalian brain. This type of imaging will be important in studying post-stroke interventions, particularly drugs that are known to alter energy metabolism.

Feeding a Saccharomyces cerevisiae fermentation product before and during a feed restriction challenge on milk production, plasma biomarkers, and immune function in Holstein cows.

Periods of decreased feed intake may disrupt function of the intestinal barrier. Feeding NutriTek® (NTK; Diamond V, Cedar Rapids, IA), a postbiotic from S. cerevisiae fermentation (SCFP), improved health and supported anti-inflammatory functions. We investigated the effects of feeding NTK to cows before and during a period of feed restriction (FR) designed to model periods of intestinal barrier dysfunction. In total, 16 multiparous cows (97.1 ± 7.6 DIM; n = 8/group) were fed a control diet (CON) or CON plus 19 g/d NTK for 9 wk (Phase 1; P1) and then were subjected to an FR challenge for 5 d, during which they were fed 40% of their ad libitum intake from the 7 d prior to FR. Milk yield (MY) and DMI were collected daily. During FR, milk was collected daily for composition, blood daily to measure plasma biomarkers and to measure monocyte and neutrophil phagocytosis and oxidative burst on d 1, 3, and 5. Data were analyzed using a mixed model in SAS 9.4. All data were subjected to repeated measures ANOVA. Dietary treatment (TRT), Day, and their interaction (TRT × Day) were considered as fixed effects and cow as the random effect. For analysis of P1, data collected during a 7-d adaptation phase were used as a covariate. During P1, NTK cows tended to have greater DMI and had greater fat, ECM and FCM yields, and feed efficiency (ECM/DMI and FCM/DMI). Protein yield tended to be greater in NTK compared with CON cows. A tendency for greater monocyte phagocytosis was detected with NTK. However, during FR, feeding NTK led to lower MY and lactose yield and tended to lower solids percentage. While NTK cows tended to have reduced neutrophil oxidative burst than CON cows during FR (NTK: 26.20%, CON: 36.93%), there was no difference in phagocytosis (NTK: 7.92%, CON: 6.31%). Plasma biomarkers of energy metabolism, liver function, inflammation, and oxidative stress during the FR period did not differ. Overall, results suggested that feeding NTK increased the yield of FCM, ECM, feed efficiency and milk components prior to FR.

Serum concentrations of folic acid and cobalamin and energy metabolism of ewes as a function of the energy density of the diet, peripartum period, and pregnancy toxemia.

The aim of this study was to determine the serum levels of folic acid, cobalamin, and indicators of energy metabolism during the peripartum period of ewes submitted to different diets and with or without pregnancy toxemia. Forty Santa Inês ewes were divided into three groups, a group of 20 animals fed only roughage-diet with low energy density (G1), a group of 10 ewes submitted to diet containing roughage and concentrate-diet with high energy density (G2), and a group of 10 ewes with diagnosis of pregnancy toxemia (PT) (G3). In the ewes of groups G1 and G2, blood samples were collected -30, -15, -7days pre-partum, on the day of partum, and 7, 15, and 30days postpartum; blood samples from G3 animals were collected during the veterinary clinic care. Significant variations were observed in the plasma concentration of glucose and serum folic acid, cobalamin, fructosamine, NEFA, and β-hydroxybutyrate in the groups fed diets with different energy levels, with and without PT (P < 0.05). A positive correlation of folic acid and cobalamin was observed with plasma glucose and serum NEFA concentrations, in addition to negative correlation of glucose with NEFA and β-hydroxybutyrate and positive correlation between the latter biomarkers of energy metabolism. The ewes diagnosed with PT showed higher levels of folic acid, NEFA, and β-hydroxybutyrate (P < 0.05). Ewes fed diet containing roughage and concentrate present higher serum levels of folic acid, cobalamin, and glucose, in the pre-partum period and in the lactation period. The results obtained contribute to better understanding of the application of blood concentration tests for folic acid and cobalamin in the energy metabolism and in PT in ewes.

UPLC-MS-based plasma metabolomics for identifying energy metabolism biomarkers of maintenance in growing pigs.

The purpose of this study is to explore the potential plasma metabolism biomarkers reflecting the maintenance status of growing pigs. The repeated measurement design was used in this experiment, and six barrows (28.6 ± 0.5 kg BW) were selected and kept in metabolism crates. The feeding level in growing pigs close to ad libitum was 2400 kJ ME/kg BW0.6 ·day-1 during Day 1 to Day 7, while a feeding level of 782 kJ ME/kg BW0.6 ·day-1 was provided as energy requirement for maintenance during Day 8 to Day 14. Plasma samples of each pig were collected from the anterior vena cava on the morning of Day 8 and Day 15. The metabolites of plasma were determined by high-resolution mass spectrometry using a metabolomics approach. Results showed that metabolomics analysis between ad libitum-fed state and maintained status revealed differences in 16 compounds. Identified compounds were enriched in metabolic pathways related to linoleic acid metabolism, tryptophan metabolism, and alanine, aspartate and glutamate metabolism. In conclusion, linoleic acid metabolism, tryptophan metabolism, alanine, aspartate and glutamic acid metabolism pathways played a major regulatory role in the maintenance status of growing pigs. The potential metabolism biomarkers of maintenance in growing pigs were linoleic acid, glutamine and tyrosine.

The promising role of probiotics/prebiotics/synbiotics in energy metabolism biomarkers in patients with NAFLD: A systematic review and meta-analysis.

BackgroundNonalcoholic fatty liver disease (NAFLD) is a chronic liver disease with a high prevalence worldwide, seriously harming human health, and its pathogenesis remains unclear. In recent years, increasing evidence has indicated that intestinal microbiota plays an important role in the occurrence and development of NAFLD. The regulation method of probiotics/prebiotics/synbiotics can alter the intestinal microbiota and has been suggested as an option in the treatment of NAFLD.MethodsFive databases of PubMed, Embase, the Cochrane Library, clinicaltrails.gov, and China National Knowledge Infrastructure were searched initially, and then the eligible studies were screened. Finally, the data of included studieswere extracted, combined and analyzedResultsA total of 29 randomized controlled trials involving 2,110 patients were included in this study. The results showed that using probiotics/prebiotics/synbiotics in the intervention group could reduce the levels of glucose (SMD = −0.23, 95% CI [−0.45, −0.01], P = 0.04), HOMA-IR (SMD = −0.47, 95% CI [−0.63, −0.31], P < 0.00001) and insulin (SMD = −0.46, 95% CI [−0.76, −0.16], P = 0.002) in sugar metabolism; in terms of lipid metabolism, the levels of TC (SMD = −0.62, 95%CI [−0.87, −0.36], P < 0.00001), and LDL-C (SMD = −0.57, 95%CI [−0.85, −0.28], P < 0.00001) were decreased; and the level of ALB was decreased in protein metabolism (SMD = −0.34, 95%CI [−0.61, −0.06], P = 0.02).ConclusionsBased on the current evidence, probiotics/prebiotics/synbiotics may improve energy metabolism biomarkers in the NAFLD population, but these effects still need to be confirmed by further research.Systematic Review Registrationhttps://www.crd.york.ac.uk/PROSPERO/#aboutpage.

Exposure to the Insecticide Sulfoxaflor Affects Behaviour and Biomarkers Responses of Carcinus maenas (Crustacea: Decapoda).

Simple SummarySulfoxaflor is an insecticide for which there are few studies regarding its toxicity to non-target organisms. The present study aimed to investigate the acute and sub-lethal effects of sulfoxaflor on Carcinus maenas by addressing survival, behaviour, and biomarkers. Sulfoxaflor affected feed intake and motricity of C. maenas. From the integrated analysis of endpoints, with the increase in concentrations of sulfoxaflor, after seven days, one can notice a lower detoxification capacity, higher lipid peroxidation, and higher motricity effects and lower feed intake. This study aims to contribute to the understanding of the negative impacts of sulfoxaflor on green crabs and increase knowledge of this pesticide toxicity to non-target coastal invertebrates.Sulfoxaflor is an insecticide belonging to the recent sulfoximine class, acting as a nicotinic acetylcholine receptor (nAChRs) agonist. There are few studies regarding sulfoxaflor’s toxicity to non-target organisms. The present study aimed to investigate the acute and sub-lethal effects of sulfoxaflor on Carcinus maenas by addressing survival, behaviour (feed intake and motricity), and neuromuscular, detoxification and oxidative stress, and energy metabolism biomarkers. Adult male green crabs were exposed to sulfoxaflor for 96 h and an LC50 of 2.88 mg L−1 was estimated. All biomarker endpoints were sampled after three (T3) and seven (T7) days of exposure and behavioural endpoints were addressed at T3 and day six (T6). Sulfoxaflor affected the feed intake and motricity of C. maenas at T6. From the integrated analysis of endpoints, with the increase in concentrations of sulfoxaflor, after seven days, one can notice a lower detoxification capacity (lower GST), higher LPO levels and effects on behaviour (higher motricity effects and lower feed intake). This integrated approach proved to be valuable in understanding the negative impacts of sulfoxaflor on green crabs, while contributing to the knowledge of this pesticide toxicity to non-target coastal invertebrates.

Long-term ovarian hormone deprivation alters functional connectivity, brain neurochemical profile and white matter integrity in the Tg2576 amyloid mouse model of Alzheimer's disease

Premenopausal bilateral ovariectomy is considered to be one of the risk factors of Alzheimer's disease (AD). However, the underlying mechanisms remain unclear. Here, we aimed to investigate long-term neurological consequences of ovariectomy in a rodent AD model, TG2576 (TG), and wild-type mice (WT) that underwent an ovariectomy or sham-operation, using in vivo MRI biomarkers. An increase in osmoregulation and energy metabolism biomarkers in the hypothalamus, a decrease in white matter integrity, and a decrease in the resting-state functional connectivity was observed in ovariectomized TG mice compared to sham-operated TG mice. In addition, we observed an increase in functional connectivity in ovariectomized WT mice compared to sham-operated WT mice. Furthermore, genotype (TG vs. WT) effects on imaging markers and GFAP immunoreactivity levels were observed, but there was no effect of interaction (Genotype × Surgery) on amyloid-beta-and GFAP immunoreactivity levels. Taken together, our results indicated that both genotype and ovariectomy alters imaging biomarkers associated with AD.

Ameliorative Impact of Aloe vera on Cartap Mediated Toxicity in the Brain of Wistar Rats.

Exposure to pesticides can pose a greater threat to multiple organs of nontarget animals. Cartap is a thiocarbamate pesticide, broadly used in agricultural fields. The assessment of neurotoxicity of cartap has not been properly studied in the mammalian systems. The present investigation unveils the toxic effects of cartap in the brain of Wistar rats its amelioration by using aqueous extract of Aloe vera leaves. We have used 4 groups of animals comprising six in each: Group 1- control, Group 2- control with Aloe vera, Group 3- cartap, Group 4- cartap with Aloe vera treated. After 15days of treatment, biochemical investigations were conducted. Wistar rats orally exposed to sublethal doses of cartap, showed significant variations in the levels of prooxidants i.e. MDA and GSH (an oxidative stress marker) and enzymatic antioxidants i.e. SOD, CAT, GST, GPx. The decreased levels of CAT, SOD, GST and increased levels of GPx were detected in the experimental rats treated with cartap. The significant alterations were recorded with the declined activities of LDH and AChE, considered as the biomarker of energy metabolism and altered cholinergic function, respectively. However, the pre-administration of aqueous extract of Aloe vera leaves was found to markedly ameliorate the toxic effects of cartap by shielding the levels of aforesaid oxidative markers near to the control. The ameliorative impact of Aloe vera, might be due to the presence of several antioxidant molecules in it which were able to counter the oxidative stress generated by cartap stress. These results suggested that Aloe vera could be utilized as a possible supplement with the relevant therapeutics in the suitable management of cartap toxicity in association.

Suitability of rumination time during the first week after calving for detecting metabolic status and lactation performance in simmental dairy cows: a cluster-analytic approach

An unsupervised machine learning approach (ML) of rumination time (RT) data was used to evaluate metabolic and lactation performance in a cohort of Simmental dairy cows (29) around parturition. The k-means clustering (unsupervised ML) was used to generate clusters based on RT (Hr-Tags; SCR by Allflex) over 24 h from 1 to 7-d post-calving. Two large clusters were obtained: high (High-RT, n = 13) and low (Low-RT, n = 12). Milk production was recorded until 42 d in lactation and milk samples collected weekly. Blood samples were collected from −21 to +28 d relative to calving. Data were subjected to PROC MIXED of SAS. Compared with Low-RT, High-RT cows had lower plasma BHB (3, 7, and 14 d), haptoglobin (3 and 7 d), and non-esterified fatty acids (1 and 3 d). High-RT cows had greater tocopherol from 3 to 28 d, fructosamine and albumin at 7 d, and retinol at 3 and 7 d after calving than Low-RT cows. Compared with multiparous Low-RT, milk production was greater in multiparous High-RT cows, but no differences were detected for primiparous. Only for multiparous, High-RT cows displayed a greater LFI than Low-Rt cows. The close relationship between RT and the physiological state at the onset of lactation supports the use of RT as an indicator of metabolic and inflammatory adaptations to the negative energy balance of cows after parturition. At the farm level, these outcomes provide information to farmers that can be helpful in management decisions for cow health, complementing traditional methods. HIGHLIGHTS The unsupervised ML applied was able to group cows with different RT increase rates after calving based on differences in plasma biomarkers of energy metabolism, inflammatory response, and liver functionality, particularly in multiparous cows. A quicker increase in RT after calving was associated with a lower inflammatory response, lower lipid mobilisation, and greater milk production. At the farm level, the fine-tuning of specific algorithm in the actual sensors considering the rate of increase of RT after calving can be helpful in management decisions for cow health, complementing traditional methods to better monitor early lactation dairy cows.

Low-Dose Ethanol Has Impacts on Plasma Levels of Metabolites Relating to Chronic Disease Risk in SAMP8 mice.

The effects of low-dose alcohol on experimental animals are unclear. This study examined plasma metabolites in senescence-accelerated mice 8 (SAMP8) given low-dose ethanol, and compared them with aging progress and skeletal muscle strength. Male SAMP8 mice (10-wk-old) were given drinking water containing 0% (control), 1%, 2%, or 5% (v/v) ethanol for 14 wk. Compared with the control group, only mice who consumed 1% ethanol experienced a lower senescence score at 18 and 23 wk, as well as an increased limb grip strength at 21 wk. Plasma metabolites of control, 1% and 2% ethanol groups were analyzed by capillary electrophoresis-time-of-flight mass spectrometry (CE-TOF/MS). Among the 7 metabolites affected by ethanol, notewhorthy is the positive association of the ethanol levels in drinking water with the levels of α-ketoglutarate (antioxidant and anti-inflammatory metabolite) and hippurate (antioxidant and microbial co-metabolite) (p<0.05). Intriguingly, the levels of 2-hydroxyisobutyrate (the biomarker of energy metabolism and microbial co-metabolite) were higher in the 1% ethanol group (p<0.05), but not in the 2% ethanol group as compared to the control. Furthermore, the levels of some of the metabolites affected were correlated with some variables in the grading score of senescence and muscle strength. This study provides a novel insight into how low-dose ethanol in SAMP8 mice modulates the levels of circulating metabolites relating to chronic disease risk.

Effects of resveratrol on alterations in cerebrum energy metabolism caused by metabolites accumulated in type I citrullinemia in rats.

We investigated the in vitro effects of citrulline (0.1, 2.5 and 5.0mM) and ammonia (0.01, 0.1 and 1.0mM), and the influence of resveratrol (0.01mM, 0.1mM and 0.5mM) on pyruvate kinase, citrate synthase, succinate dehydrogenase (SDH), complex II, and cytochrome c oxidase activities in cerebral cortex, cerebellum and hippocampus homogenates of 60-day-old male Wistar rats. Results showed that 2.5 and 5.0mM citrulline decreased pyruvate kinase activity in cerebral cortex and, at a concentration of 5.0mM, increased its activity in hippocampus. Additionally, 5.0mM citrulline increased citrate synthase activity in the cerebellum of rats. Citrulline (5.0mM) reduced complex II and cytochrome c oxidase activities in cerebral cortex and hippocampus. With regard to ammonia, at 0.1 and 1.0mM, decreased complex II activity in cerebral cortex and at 1.0mM decreased its activity in cerebellum and hippocampus. Ammonia (1.0mM) also decreased cytochrome c oxidase activity in cerebral cortex and cerebellum of rats. Resveratrol was able to prevent most of the alterations caused by these metabolites in the biomarkers of energy metabolism measured in the cerebrum of rats. Data suggest that these alterations in energy metabolism, caused by citrulline and ammonia, are probably mediated by the generation of free radicals, which can in turn be scavenged by resveratrol.

Effects of Lonicera japonica extract on performance, blood biomarkers of inflammation and oxidative stress during perinatal period in dairy cows

ObjectiveAn experiment was conducted to evaluate the effects of Lonicera japonica extract (LJE) on milk production, rumen fermentation and blood biomarkers of energy metabolism, inflammation and oxidative stress during the perinatal period of Holstein dairy cows.MethodsEighteen Holstein dairy cows were used in a complete randomized design experiment with 3 dietary treatments and 6 cows per treatment. All cows received the same basal total mixed ration (TMR) including a prepartal diet (1.35 Mcal of net energy for lactation [NEL]/kg of dry matter [DM], 13.23% crude protein [CP]) from −60 d to calving and a postpartal diet (1.61 Mcal of NEL/kg of DM, 17.39% CP) from calving to 30 days in milk (DIM). The 3 dietary treatments were TMR supplemented with LJE at 0 (control), 1 and 2 g/kg DM, respectively. LJE was offered from 21 d before calving to 30 DIM. Dry matter intake (DMI) and milk production were measured daily after calving. Milk and rumen fluid samples were collected on 29 and 30 d after calving. On −10, 4, 14, and 30 d relative to calving, blood samples were collected to analyze the biomarkers of energy metabolism, inflammation and oxidative stress.ResultsCompared with control diet, LJE supplementation at 1 and 2 g/kg DM increased DMI, milk yield and reduced milk somatic cell count. LJE supplementation also decreased the concentrations of blood biomarkers of pro-inflammation (interleukin-1β [IL-1β], IL-6, and haptoglobin), energy metabolism (nonesterified fatty acid and β-hydroxybutyric acid) and oxidative stress (reactive oxygen metabolites), meanwhile increased the total antioxidant capacity and superoxide dismutase concentrations in blood. No differences were observed in rumen pH, volatile fatty acid, and ammonia-N (NH3-N) concentrations between LJE supplemented diets and the control diet.ConclusionSupplementation with 1 and 2 g LJE/kg DM could increase DMI, improve lactation performance, and enhance anti-inflammatory and antioxidant capacities of dairy cows during perinatal period.

Effects of triclosan on early development of Solea senegalensis: from biochemical to individual level

Harmful effects of triclosan (TCS) have been reported on several organisms; however, effects on early life stages of marine vertebrates are limited. Therefore, the objective of this work was to assess the effects of TCS during early development of the flatfish Solea senegalensis after initial characterization of cholinesterases (ChEs) and determination of selected biochemical markers baseline levels.Characterization of ChEs and determination of biochemical markers baseline levels of cholinergic activity, energy metabolism and oxidative stress were analysed in sole at 3 days after hatching (dah) and at the onset and end of metamorphosis. To assess TCS effects, fish were exposed during 96h to 30-500 μg L−1 TCS until 3 dah. Fish at 13 dah were exposed during 48h to 200-1,500 μg L−1 TCS and maintained until complete metamorphosis. Effects on survival, malformations, length, metamorphosis progression and biochemical markers were evaluated.The main ChE active form present in sole early life stages is acetylcholinesterase and baseline levels of oxidative stress and energy metabolism biomarkers changed according to fish developmental stage. Triclosan induced malformations (EC50 = 180 μg L−1 at 3 dah), decreased growth (95 μg L−1 at 3 dah; 548 μg L−1 at 24 dah) and affected metamorphosis progression (391 μg L−1 at 17 dah). Impairment of antioxidant system was observed, with TCS affecting catalase at the end of metamorphosis test, however, no oxidative damage on lipids was detected. Glutathione S-transferase was the most sensitive endpoint during early larval test (LOEC = 30 μg L−1).Exposure to TCS affected S. senegalensis at individual and sub-individual levels, both at early larval stage and during the critical period of metamorphosis.

Toxicity of the insecticides spinosad and indoxacarb to the non-target aquatic midge Chironomus riparius

Spinosad and indoxacarb are two relatively new insecticides mainly used in agriculture to control insect pests. However, at their current application rates, non-target aquatic insect species may also be impacted. In this study, larvae of the non-biting midge Chironomus riparius were exposed in the laboratory to both insecticides and their effects evaluated at the organismal level, using standard ecotoxicological tests, and at the biochemical level, by monitoring specific oxidative stress, neuronal, and energy metabolism biomarkers. Chronic exposure to both insecticides compromised growth and emergence of C. riparius. Short-term exposures revealed alterations at biochemical level that might be related to the toxicological targets of both insecticides. Growth and development time were the most sensitive endpoints at individual level for both pesticides, while at the biochemical level, the electron transport system activity was the most sensitive biomarker for spinosad exposure, suggesting an increase in energy demands associated with the activation of defense mechanisms. Glutathione-S-transferase was the most sensitive biomarker for indoxacarb exposure, underlining the role of this enzyme in the detoxification of indoxacarb. Additionally, changes in lactate dehydrogenase and glutathione peroxidase activities were observed for both insecticides, and evidences of oxidative damage were found for spinosad. This study contributes to the growing knowledge on sublethal effects of novel insecticides on non-target aquatic invertebrates and strengthens the usefulness of biochemical biomarkers to support the interpretation of their potentially deleterious effects on aquatic insects near agricultural fields.

Oxidative stress, energy metabolism and molecular responses of earthworms (Eisenia fetida) exposed to low-density polyethylene microplastics.

Soils are both a sink and a pathway of plastic wastes, but there is a great lack of knowledge regarding their impacts on soil biota. To tackle the mechanisms of toxicity of these contaminants to soil invertebrates, earthworms (Eisenia fetida Savigny, 1826) were exposed during 28days to different concentrations of low-density polyethylene microplastics (62, 125, 250, 500 and 1000mg MPs kg-1 soildw) with sizes ranging between 250 and 1000μm, in an artificial soil. The ecotoxicological responses were evaluated by analysing various oxidative stress biomarkers (catalase, glutathione S-transferase and thiobarbituric acid reactive substances), a biomarker of energy metabolism (lactate dehydrogenase) and overall organism molecular changes by Fourier transform infrared spectrometry (FTIR) and nuclear magnetic resonance (NMR) analyses. Significant effects resulting from an unbalanced oxidative stress system, expressed in terms of thiobarbituric acid reactive substances levels were recorded on earthworms exposed at the three highest concentrations tested. Despite that, no significant changes were recorded on the molecular profiles of earthworms by FTIR-ATR. NMR analysis pointed out for differences from the control, only for earthworms exposed to the lowest concentration of MPs. Considering that stress responses are complex, and involve multiple mechanisms, a cluster analysis taking into account all the parameters assessed, clearly identified two groups of earthworms separated by the concentration of 250mg MPs kg-1 soildw, above each meaningful effects were recorded.

Ecotoxicological evaluation of gilthead seabream (Sparus aurata) exposed to the antibiotic oxytetracycline using a multibiomarker approach

Oxytetracycline (OTC) is an antibiotic widely used in human and veterinary medicines. Since the primary toxicity occurs mainly at molecular/biochemical levels, the study of different biological responses corresponds to a sensitive and crucial approach. The aim of the present study was to assess the toxic effects of OTC in gilthead seabream (Sparus aurata) through the use of multibiomarkers and elucidate about the possible toxicological mechanisms involved. S. aurata were acutely (96 h: 0.04–400 μg/L) and chronically (28 days: 0.0004–4 μg/L) exposed to OTC. Detoxification, antioxidant defense, lipid peroxidation, genotoxicity, neurotransmission and energy metabolism biomarkers were evaluated. OTC impaired the detoxification pathways and caused peroxidative damage and genotoxicity. The relevance of the here-obtained data is high, since significant effects were recorded for levels already reported to occur in the wild, meaning that environmentally-exposed marine organisms (including those cultured at fish farms) are not completely exempt of risks posed by OTC.

Study on denervated rat skeletal muscle by 7.0 T 31P-MR phosphorus spectroscopy

Objective To investigate the changes of 31P-MRS in denervated skeletal muscle at 7.0 T MR system. Methods In the experiment group, a total of 18 male Sprague-Dawley rats aged 6-8 weeks old and weighing 200-250 g were obtained. The right posterior femoral nerve were transected, and the proximal stumps were ligated by using 5-0 nylon stitches to preclude spontaneous repair. A sham surgery (incision and exploration of the nerve) was performed at the same time (n=6). Before rat model established and at varying times after initial surgery (3 d, and 1, 2, 4, 6, and 8 weeks). 31P-MR spectra of rat quadriceps femoris were acquired on 7.0 T Agilent small animal MR imaging system. We quantified the phosphocreatine (PCr), adenosine triphosphate(β-ATP), and inorganic phosphate (Pi) using Creatine phosphate disodium salt(10 mmol/L) as an external standard. The ratios, include PCr/Pi, Pi/β-ATP, PCr/β-ATP, PCr/(PCr+Pi) and Pi/(PCr+Pi) were quantified in jMRUI. All data were analyzed by one way analysis of variance (ANOVA) with posttest inter-group comparisons using Bonferroni test. Comparative analysis methods between the experimental and control group were performed via independent samples t test. P<0.05 was considered as statistically significance. Results The average measured concentrations of β-ATP, PCr, and Pi in control groups were (6.654±0.178) μmol/g, (25.656±0.738) μmol/g, and (1.594±0.096) μmol/g, respectively. There were significant statistically differences in β-ATP, PCr, Pi, PCr/Pi, PCr/PCr+Pi,Pi/PCr+Pi between the study and control group(P<0.05) at any measurement time point after denervation. Pi/β-ATP,PCr/β-ATP and pHi in the experimental group significantly statistically differed from those of the control group (all P<0.05)at each time point except on day 3 after operation. The total concentrations of β-ATP and PCr were reduced by 11.5% and 19.7% respectively on day 3. Thereafter, β-ATP and PCr declined rapidly by 63.1% and 68.8% at week 4 respectively, then decreased slowely by 74.0%和82.3% till week 10. The change of PCr/Pi is similar to β-ATP and PCr, but more remarkable. Pi, Pi/PCr+Pi, Pi/ATP showed a progressively increase till week 10. The intra-cellular pH (pHi) of normal rat muscles was 7.033±0.017, While the pHi gently in experiment rat muscles decreased during the entire experiment. Conclusion 31P-MRS with 7.0 T can quantify the temporal changes of energy metabolism and pHi in normal and denervated rat muscles. It shows that the dysfunction of energy metabolism are progressive with time and that they begin within a short period following the nerve section, the change of β-ATP, PCr, PCr/Pi take place primarily within 4 weeks after denervation. β-ATP, PCr, and PCr/Pi may be potential biomarkers of energy metabolism in the evaluation of denervated muscle atrophy. Key words: Magnetic resonance spectroscopy; Muscular disorders, atrophic; Denervation; Energy metabolism; Intracellular pH

Plasma fructosamine during the transition period and its relationship with energy metabolism and inflammation biomarkers in dairy cows

Plasma concentration of fructosamine (Fr) has been proposed as a way to monitor retrospective undernutrition in dairy cows during the transition period. The objective of this study was to evaluate, under standard feeding conditions, the variation of plasma Fr in dairy cows during the transition phase and its relationship with biomarkers of energy metabolism and inflammatory conditions. The trial was carried out with 24 Italian Friesian dairy cows from the last month of pregnancy through the first 2 months of lactation. Diet characteristics and dry matter intake (DMI) were continuously monitored, and energy balance was evaluated on a weekly basis. Milk yield, body weight, body condition score and health status were also monitored. Blood samples were taken two times a week throughout the experimental period and daily in the week before and after calving. Concentrations of Fr together with biochemical indicators related to energy, protein and mineral metabolism; markers of inflammation; and liver enzymes were measured in plasma. Cows were categorized into three groups according to the average concentration of plasma Fr observed at 28, 35 and 42 days in milk (DIM) (cows with lowest [LF], intermediate [IF] and highest [HF] concentrations of Fr, respectively). The LF cows reached an average maximum milk yield (MY) of 51.7 kg/d, whereas IF and HF cows produced 48.9 and 47.2 kg/d, respectively. During the first month of lactation, the DMI was 17.4, 18.2 and 18.7 kg/d in LF, IF and HF, respectively. In the first month of lactation, Fr was not correlated with MY, but had a positive correlation with DMI (P < 0.05) and energy balance (P < 0.05). In the same period, the average plasma glucose was greater (P < 0.05) in HF (3.71 mmol/L) and lower in LF (3.34 mmol/L). During the first month of lactation, plasma Fr was positively related (P < 0.05) to plasma glucose and albumin concentrations over the previous 1–3 weeks. The Liver Activity Index, used to measure the consequences of inflammation around calving and calculated by using three blood analytes that represent liver-synthesized proteins, indicated greater inflammation in LF cows compared with HF. Our results highlight that plasma Fr during the first 3–4 weeks of lactation summarizes the energy balance during the transition period and that the energy deficit is more influenced by DMI and inflammatory conditions than by MY.

Autofluorescence-based optical biopsy: An effective diagnostic tool in hepatology.

Autofluorescence emission of liver tissue depends on the presence of endogenous biomolecules able to fluoresce under suitable light excitation. Overall autofluorescence emission contains much information of diagnostic value because it is the sum of individual autofluorescence contributions from fluorophores involved in metabolism, for example, NAD(P)H, flavins, lipofuscins, retinoids, porphyrins, bilirubin and lipids, or in structural architecture, for example, fibrous proteins, in close relationship with normal, altered or diseased conditions of the liver. Since the 1950s, hepatocytes and liver have been historical models to study NAD(P)H and flavins as in situ, real-time autofluorescence biomarkers of energy metabolism and redox state. Later investigations designed to monitor organ responses to ischaemia/reperfusion were able to predict the risk of dysfunction in surgery and transplantation or support the development of procedures to ameliorate the liver outcome. Subsequently, fluorescent fatty acids, lipofuscin-like lipopigments and collagen were characterized as optical biomarkers of liver steatosis, oxidative stress damage, fibrosis and disease progression. Currently, serum AF is being investigated to improve non-invasive optical diagnosis of liver disease. Validation of endogenous fluorophores and in situ discrimination of cancerous from non-cancerous tissue belong to the few studies on liver in human subjects. These reports along with other optical techniques and the huge work performed on animal models suggest many optically based applications in hepatology. Optical diagnosis is currently offering beneficial outcomes in clinical fields ranging from the respiratory and gastrointestinal tracts, to dermatology and ophthalmology. Accordingly, this review aims to promote an effective bench to bedside transfer in hepatology.