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Polyamines (PAs), including putrescine (Put), spermidine (Spd), and spermine (Spm), are essential polycations with wide-ranging roles in cellular functions. PA levels decline with age, making exogenous PA supplementation, particularly Spd, an intriguing prospect. Previous research in honey bees demonstrated that millimolar Spd added to their diet increased lifespan and reinforced oxidative resilience. The present study is aimed to assess the anti-aging effects of spermidine supplementation at concentrations of 0.1 and 1 mM in honey bees, focusing on autophagy and associated epigenetic changes. Results showed a more pronounced effect at the lower Spd concentration, primarily in the abdomen. Spd induced site-specific histone 3 hypoacetylation at sites K18 and 27, hyperacetylation at K9, with no change at K14 in the entire body. Additionally, autophagy-related genes (ATG3, 5, 9, 13) and genes associated with epigenetic changes (HDAC1, HDAC3, SIRT1, KAT2A, KAT6B, P300, DNMT1A, DNMT1B) were upregulated in the abdomens of honey bees. In conclusion, our findings highlight profound epigenetic changes and autophagy promotion due to spermidine supplementation, contributing to increased honey bee longevity. Further research is needed to fully understand the precise mechanisms and the interplay between epigenetic alterations and autophagy in honey bees, underscoring the significance of autophagy as a geroprotective mechanism. [ABSTRACT FROM AUTHOR]

Over the past decades, the number of managed honey bee Apis mellifera L. (Hymenoptera: Apidae) colonies have been decreasing. The majority of losses occur during winter, suggesting that overwintering honey bees are more susceptible to adverse factors. We focused on the oxidative status of overwintering honey bees, particularly at the beginning (November) and end (March) of the wintering period. Colonies from three locations with different anthropogenic influences were selected: Belgrade, an urban zone, Zajača, an industrial zone, and Susek, a rural area. We measured levels of malondialdehyde (MDA), as a marker of lipid peroxidation, as well as the expression and activity of select antioxidative enzymes: superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST). Our results show that enzyme activity and gene expression of antioxidative enzymes are influenced by both sample location and the time of sampling. The majority of analyzed genes had significantly reduced expression, at the end of the overwintering period when higher activities of antioxidative enzymes were also recorded. Among the analyzed parameters, SOD activity and gene expression of microsomal GST isoforms were more affected by local environmental conditions, suggesting the complex role of these enzymes in antioxidative defense and detoxification. The higher MDA levels observed at the end of overwintering for all three locations likely reflects accumulated oxidative damage which could be associated with the aging process, brood rearing and/or the onset flying activity. [ABSTRACT FROM AUTHOR]

The European corn borer (ECB, Ostrinia nubilalis Hbn.) is a major pest in temperate regions of Europe and North America. Fifth instar ECB larvae enter diapause before winter and gradually develop cold hardiness. Here we investigated the combined influence of diapause phase and low temperature on sugar and polyol content in ECB larvae. Larvae in mid-diapause or diapause termination were acclimated at 5 °C, −3°C or −16 °C, and sugar and polyol content was measured using GC–MS. Control GC–MS measurements were conducted on untreated non-diapausing larvae. We detected differences in polyol (glycerol, sorbitol, myo -inositol) and sugar (trehalose, fructose, glucose) levels in diapausing versus non-diapausing larvae. Glycerol and trehalose were the most abundant of all analyzed cryoprotective compounds in diapausing larvae. Exposure of diapausing larvae to decreasing temperatures induced changes in polyol and sugar levels that depended on the phase of diapause. In mid-diapause larvae, decreasing temperatures induced a significant increase in glycerol and a decrease in sorbitol and myo -inositol. In larvae at diapause termination, polyol content was lower and less influenced by decreasing temperatures. In contrast, sugar levels were lower in larvae at mid-diapause versus diapause termination. Exposure of larvae to −16 °C induced a significant increase in the levels of all detected sugars. In particular, glucose levels were significantly higher in larvae at diapause termination following exposure to −16 °C. We propose that this shift toward sugar synthesis following low temperature exposure in larvae at diapause termination is a consequence of NADPH dependent polyol synthesis, and may be a mechanism for preservation of carbon reserves needed for post-diapause development. [ABSTRACT FROM AUTHOR]

In this study, laboratory bioassays were performed to investigate the impact of sublethal concentrations of Cu (CuCl 2 : 1000, 100, 10 mg L −1 ), Pb (PbCl 2 : 10, 1, 0.1 mg L −1 ) and Cd (CdCl 2 : 0.1, 0.01, 0.001 mg L −1 ) on honey bee redox status and the activity of the main antioxidative enzymes and their gene expression. Our results show that exposure to these metals led to significant changes of gene expression, the levels of enzyme activity and redox status, but the effects are metal and dose dependent. In general, exposure of 48 h to given concentrations of Cu, Cd and Pb did not change the activity of antioxidative enzymes and the level of lipid peroxidation, with the exception of decreased activity of catalase at the lowest concentration of cadmium. Only lead produced increases in glutathione and thiol groups. Expression of genes for catalase and superoxide dismutase changed with exposure to cadmium and copper, whilst lead induced only expression of superoxide dismutase genes. The results from this study provide basic data for future research regarding the impacts of metal pollution on Apis mellifera and will be an important step towards a comprehensive risk assessment of the environmental stressors on honey bees. [ABSTRACT FROM AUTHOR]

In this study, an ¹H-NMR -based metabolomic approach was used to investigate the biochemical mechanisms of diapause and cold hardiness in diapausing larvae of the European corn borer Ostrinia nubilalis. Metabolomic patterns in polar hemolymph extracts from non-diapausing and diapausing larvae of O. nubilalis were compared. Analysis indicated 13 metabolites: 7 amino acids, glycerol, acetate, citrate, succinate, lactate and putrescine. Results show that diapausing larvae display different metabolomic patterns compared to active non-diapausing larvae, with predominant metabolites identified as glycerol, proline and alanine. In specific diapausing larvae initially kept at 5 °C then gradually chilled to -3 °C and -16 °C, alanine, glycerol and acetate were predominant metabolites. ¹H-NMR spectroscopy provides new insight into the metabolomic patterns associated with cold resistance and diapause in O. nubilalis larvae, suggesting distinct metabolomes function in actively developing and diapausing larvae. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the effect of dietary supplements of lead acetate and aluminosilicates on oxidative status in brain, kidney and duodenum of broiler chickens. In this regard, activity of antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPx), pyrogallol peroxidase (PPx) and lipid peroxidation was determined in examined organs. The experiment was performed with eighty-four broiler chickens of both sexes randomly allotted to four diets including the control group, the Pb group containing 500 mg lead acetate/kg diet, the ATN (Antitoxic nutrient) group with 5 g mixture of aluminosilicates (zeolite and montmorillonite)/kg diet and the Pb + ATN group containing 500 mg lead acetate and 5 g mixture of aluminosilicates/kg diet, in a 3-week feeding experiment. Each of these groups consisted of seven replications with 3 chickens per replication. Results suggest that aluminosilicates alone did not provoke any adverse effects and did not disturb normal biochemical and physiological homeostasis in the broilers. The dietary intake of lead acetate induced oxidative stress and promoted increase in level of malondialdehid (MDA), a lipid peroxidation marker, in all examined organs. The lead intake induced increase in CAT activity in all organs, while it decreased the activity of GPx and PPx in the brain and duodenum but did not affect that in the kidneys. The activity of SOD increased in the kidneys and duodenum but did not significantly change in the brain. The combined data showed that the chickens fed aluminosilicates received significant protection against the effects of lead acetate for most parameters measured which remained at control level. [ABSTRACT FROM AUTHOR]

This study investigated the effects of dietary supplementation with hydrated aluminosilicate (antitoxic nutrient-ATN), based on zeolitic ore (of > 90% clinoptilolite), bentonite (of > 83% montmorillonite), and small amounts of activated charcoal, on performance, hematological, serum, and liver biochemical parameters, as well as organ weights and meat quality in broiler chickens. The study included 300 1-day-old broilers of both sexes, which were assigned at random to 2 groups based on treatment: no dietary supplementation (control) and supplementation with 5 g/kg of ATN (ATN group). Dietary supplementation with 5 g/kg of ATN in broiler diets significantly increased weight gain only during the 1st and 3rd weeks; however, the effect on weight gain and the feed conversion ratio was not significant (P > 0.05) for the overall study period. Most hematological, serum, and liver biochemical indices were unaffected by the dietary treatment. Supplemental ATN significantly increased serum amylase and lactate dehydrogenase activity (P < 0.05). Spleen, proventriculus, ventriculus, and ileum weights were significantly higher (P < 0.05) in the ATN group, whereas the weights of the other measured organs were not affected by the dietary treatment. Significantly higher (P < 0.05) protein and ash, and significantly lower fat content was observed in the breast meat of chicks in the ATN group. Ash content was also significantly higher (P < 0.05) in drumstick meat. The results of this study demonstrate that supplementation with 5 g/kg of hydrated aluminosilicate influenced serum traits, organ weights, and the chemical composition of broiler chicken meat. [ABSTRACT FROM AUTHOR]

Acrylamide (AA) toxicity is associated with oxidative stress. During detoxification, AA is either coupled to gluthatione or biotransformed to glycidamide by the enzyme cytochrome P450 2E1 (CYP2E1). The aim of our study was to examine the hepatotoxicity of AA in vivo and in vitro. Thirty male Wistar rats were treated with 25 or 50 mg/kg b.w. of AA for 3 weeks. Qualitative and quantitative immunohistochemical evaluation of inducible nitric oxide synthase (iNOS), CYP2E1, catalase (CAT), superoxide dismutase 1 (SOD1), and SOD2 expression in liver was carried out. Bearing in mind that the liver is consisted mainly of hepatocytes, in a parallel study, we used the rat hepatoma cell line H4IIE to investigate the effects of AA at IC20 and IC50 concentrations on the redox status and the activity of CAT, SOD, and glutathione-S-transferase (GST), their gene expression, and CYP2E1 and iNOS expression. Immunohistochemically stained liver sections showed that treatment with AA25mg induced a significant decrease of CYP2E1 protein expression (p < 0.05), while treatment with AA50mg led to a significant increase of iNOS protein expression (p < 0.05). AA treatment dose-dependently elevated SOD2 protein expression (p < 0.05), while SOD1 protein expression was significantly increased only at AA50mg (p < 0.05). CAT protein expression was not significantly affected by AA treatments (p > 0.05). In AA-treated H4IIE cells, a concentration-dependent significant increase in lipid peroxidation and nitrite levels was observed (p < 0.05), while GSH content and SOD activity significantly decreased in a concentration-dependent manner (p < 0.05). AA IC50 significantly enhanced GST activity (p < 0.05). The level of mRNA significantly increased in a concentration-dependent manner for iNOS, SOD2, and CAT in AA-treated H4IIE cells (p < 0.05). AA IC50 significantly increased the transcription of SOD1, GSTA2, and GSTP1 genes (p < 0.05), while AA IC20 significantly decreased mRNA for CYP2E1 in H4IIE cells (p < 0.05). Obtained results indicate that AA treatments, both in vivo and in vitro, change hepatocytes; drug-metabolizing potential and disturb its redox status. [ABSTRACT FROM AUTHOR]

Dietary supplementation has been proposed as a sustainable way to improve the health and resilience of honey bees (Apis mellifera , L.), as the decline in their numbers in recent decades has raised scientific, environmental, and economic concerns. Spermidine, a natural polyamine, has been shown to be a promising substance for honey bee supplementation, as its health-promoting effects have been demonstrated in numerous studies and in different organisms. As already shown, supplementation with spermidine at a certain concentration prolonged lifespan, reduced oxidative stress, and increased antioxidative capacity in honey bees. The aim of the present study was to investigate whether spermidine supplementation affects gene expression and/or enzyme activity of antioxidative and detoxification enzymes and immune response markers in honey bee workers. The different gene expression and enzyme activity patterns observed in abdominal and head tissues in response to spermidine supplementation suggest tissue-specific and concentration-dependent effects. In addition, the immune response markers suggest that spermidine has the ability to boost honey bee immunity. The observed changes make a valuable contribution to understanding the molecular mechanisms by which spermidine may exert its beneficial effects on the bee's health and lifespan. These results support the idea of the use of spermidine supplementation to promote bee health and resilience to environmental stressors, emphasizing that the dose must be carefully chosen to achieve a balance between the pro- and antioxidant effects of spermidine. [ABSTRACT FROM AUTHOR]

Flooding stress nowadays is one of the major stressors for plants under climate change. This kind of stress may cause severe depression of the plant's growth through inhibition of photosynthesis and oxidative cell damage as well as changes in cell respiration. The present work aimed to study the effect of flooding stress on oxidative and antioxidative parameters in leaves of two maize hybrids (ZP 555 and ZP 606). Leaves of maize plants at the stage of three fully developed leaves were harvested after 6, 24, 72, and 144 h of applied flooding stress. Leaves were used for determination of physiological (the content of photosynthetic pigments and soluble proteins), oxidative stress parameters (the content of malondialdehyde (MDA) and H2O2) as well as antioxidants (the total polyphenols content, and activity of antioxidative enzymes [catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1), and Class III peroxidases (POX, EC, 1.11.1.7)]). Results indicated that flooding stress-induced time-dependent changes of measured parameters and those hybrids differ in response to stress. The noticeable difference between hybrids was detected in the H2O2 and MDA content. An increase in the activity of SOD, POX and polyphenols content, with the most pronounced changes in POX activity and polyphenols concentration, could minimize the cellular damage caused by flooding. The results of the present study suggest that a more robust antioxidative metabolism is essential under flooding stress and could be a protective strategy against oxidative damage induced by flooding in ZP 606 maize plants compared to ZP 555 plants. [ABSTRACT FROM AUTHOR]

The European corn borer (Ostrinia nubilalis, Hbn.), enters diapause, a strategy characterized by arrest of development and reproduction, reduction of metabolic rate and the emergence of increased resistance to challenging seasonal conditions as low sub-zero winter temperatures. The aim of this study was to investigate the potential role of inorganic elements in the ecophysiology of O. nubilalis, analysing their content in the whole body, hemolymph and fat body, both metabolically active, non-diapausing and overwintering diapausing larvae by ICP-OES spectrometer following the US EPA method 200.7:2001. O nubilalis as many phytophagous lepidopteran species maintain a very low extracellular sodium concentration and has potassium as dominant cation in hemolymph of their larvae. Changes in hemolymph and the whole body sodium content occur already at the onset of diapause (when the mean environmental temperatures are still high above 0 ºC) and remain stable during the time course of diapause when larvae of this species cope with sub-zero temperatures, it seems that sodium content regulation is rather a part of diapausing program than the direct effect of exposure to low temperatures. Compared to non-diapausing O. nubilalis larvae, potassium levels are much higher in the whole body and fat body of diapausing larvae and substantially increase approaching the end of diapause. The concentration of Ca, Mg, P and S differed in the whole body, hemolymph and fat body between non-diapausing and diapausing larvae without a unique trend during diapause, except an increase in their contents at the end of diapause. [ABSTRACT FROM AUTHOR]

This study was undertaken in order to investigate the biological activity of chemically characterized Carum carvi essential oil (EO) and its effect on oxidative stress parameters in stored product pests. Gas chromatography-mass spectrometry depicted carvone (68.22 ± 0.62%) and limonene (21.80 ± 0.54%) as the major compounds of said EO. Different concentrations of EO were tested for fumigant activity and showed prominent results following a timeresponse relationship in both tested species. The changes in the antioxidative defense system, catalase (CAT), superoxide dismutase (SOD), and glutathione-S-transferase (GST) activities, as well as the level of lipid peroxidation (MDA) and the content of reduced glutathione (GSH) were estimated in time-dependent response. The results demonstrated that C. carvi could have a significant role in the future, as an environmentally friendly alternative for the management of stored product pests. [ABSTRACT FROM AUTHOR]

Metallothioneins are ubiquitous proteins important in metal homeostasis and detoxification. However, they have not previously been identified in honey bees or other Hymenoptera, where metallothioneins could be of ecophysiological and ecotoxicological significance. Better understanding of the molecular responses to stress induced by toxic metals could contribute to honey bee conservation. In addition, honey bee metallothionein could represent a biomarker for monitoring environmental quality. Here we identify and characterize a metallothionein gene in Apis mellifera (AmMT). AmMT is 1,680 bp long and encodes a 48 amino acids protein with 15 cysteines and no aromatic residues. A metal response element upstream of the start codon, coupled with numerous cis‐regulatory elements indicate the functional context of AmMT. Molecular modelling predicts several transition metal binding sites, and comparative phylogenetic analysis revealed five putative metallothionein proteins in three other hymenoptera species. AmMT was characterized by cloning the full‐length coding sequence of the putative metallothionein. Recombinant AmMT was found to increase metal tolerance upon overexpression in Escherichia coli supplemented with Cd, Cu or Pb. Finally, in laboratory tests on honey bees, gene expression profiles showed a dose‐dependant relationship between Cd, Cu and Pb concentrations present in food and AmMT expression, while field experiments showed induction of AmMT in bees from an industrial site compared to those from an urban area. These studies suggest that AmMT has metal binding properties in agreement with a possible role in metal homeostasis. Further functional and structural characterization of metallothionein in honey bees and other Hymenoptera are necessary. [ABSTRACT FROM AUTHOR]

Oxidative stress is one of the principle mechanism of acrylamide-induced toxicity. Acrylamide is metabolized by cytochrome P450 2E1 (CYP2E1) to glycidamide or by direct conjugation with glutathione. Bearing in mind that up to now the effects of acrylamide on oxidative stress status and CYP2E1 level in endocrine pancreas have not been studied we performed qualitative and quantitative immunohistochemical evaluation of inducible nitric oxide synthase (iNOS), superoxide dismutase 1 (SOD1), superoxide dismutase 2 (SOD2), catalase (CAT) and CYP2E1 expression in islets of Langerhans of rats subchronically treated with 25 or 50 mg/kg bw of acrylamide. Since the majority of cells (>80%) in rodent islets are beta cells, in parallel studies, we employed the Rin-5F beta cell line to examine effects of acrylamide on redox status and the activity of CAT, SOD and glutathione-S-transferase (GST), their gene expression, and CYP2E1, NF-E2 p45-related factor 2 (Nrf2) and iNOS expression. Immunohistochemically stained pancreatic sections revealed that acrylamide induced increase of iNOS and decrease of CYP2E1 protein expression, while expression of antioxidant enzymes was not significantly affected by acrylamide in islets of Langerhans. Analysis of Mallory-Azan stained pancreatic sections revealed increased diameter of blood vessels lumen in pancreatic islets of acrylamide-treated rats. Increase in the GST activity, lipid peroxidation and nitrite level, and decrease in GSH content, CAT and SOD activities was observed in acrylamide-exposed Rin-5F cells. Level of mRNA was increased for iNOS, SOD1 and SOD2, and decreased for GSTP1, Nrf2 and CYP2E1 in acrylamide-treated Rin-5F cells. This is the first report of the effects of acrylamide on oxidant/antioxidant parameters and CYP2E1 expression in pancreatic endocrine cells. [ABSTRACT FROM AUTHOR]

Honey bee health has been an important and ongoing topic in recent years. Honey bee is also an important model organism for aging studies. Polyamines, putrescine, spermidine and spermine, are ubiquitous polycations, involved in a wide range of cellular processes such as cell growth, gene regulation, immunity, and regulation of lifespan. Spermidine, named longevity elixir, has been most analysed in the context of aging. One of the several proposed mechanisms behind spermidine actions is antioxidative activity. In present study we showed that dietary spermidine supplementation: (a) improved survival, (b) increased the average lifespan, (c) influenced the content of endogenous polyamines by increasing the level of putrescine and spermidine and decreasing the level of spermine, (d) reduced oxidative stress (MDA level), (e) increased the antioxidant capacity of the organism (FRAP), (f) increased relative gene expression of five genes involved in polyamine metabolism, and (g) upregulated vitellogenin gene in honey bees. To our knowledge, this is the first study on honey bee polyamine levels in reference to their longevity. These results provide important information on possible strategies for improving honey bee health by introducing spermidine into their diet. Here, we offer spermidine concentrations that could be considered for that purpose. [ABSTRACT FROM AUTHOR]

The honey bee (Apis mellifera L.), as an eusocial insect species, is an important model organism in research focusing on ageing and longevity, due to prominent seasonal lifespan plasticity within the worker caste (summer and winter worker bees). In this study, we employed a screening approach to evaluate several molecular parameters, providing comprehensive insights into the antioxidative (superoxide dismutase and catalase activity, reduced glutathione and sulfhydryl group content, total antioxidative capacity), detoxifying (glutathione S -transferase and acetylcholinesterase activity), and immune (phenol oxidase and glucose oxidase activity) status, as well as vitellogenin content, in the summer and winter generation of honey bees, across ageing stages and in two body compartments: the whole abdomen and the head. Summer worker bees were collected weekly for six weeks, while winter bees were collected monthly for five months. The results of our study clearly indicate a reduced overall antioxidative capacity of older groups of worker bees from both generations, while the parameters of immune responsiveness mostly contributed to the separation between the two generations based on season rather than age categories. Detoxification ability appeared to be more susceptible to environmental factors. An age-dependent increase in vitellogenin content was recorded in the abdomen, but without seasonal differences. These findings provide an excellent starting point for further investigations into age-related changes, particularly within the context of honey bee sociality. [Display omitted] • Overall antioxidative capacity is reduced with ageing in summer and winter bees. • Immune responsiveness of bees (proPOx, POx and GOx) showed seasonal variations. • Detoxification ability was more susceptible to environmental factors, not ageing. • Vitellogenin content increased in the abdomen with ageing, but without seasonal differences. [ABSTRACT FROM AUTHOR]

The use of insecticides, especially neonicotinoids (NEOCs), poses a significant threat to honey bees, thus prompting the EU to ban all outdoor uses of clothianidin, imidacloprid, and thiamethoxam. However, due to the persistence of NEOCs, as well as the fact that certain NEOCs are in use in some areas of the world, exposure to them is still an on-going issue. In the present study, we used laboratory acute toxicity test to examine the effects of sublethal concentrations (resulting in consumed doses approximately 50, 20, and 5 times below LD50) of thiacloprid and clothianidin on selected parameters of antioxidative defense (superoxide dismutase, catalase, glutathione S-transferase activities), oxidative status (reduced glutathione, protein thiols, malondialdehyde), neurotoxicity (acetylcholinesterase activity), and immune response (prophenoloxidase and phenoloxidase activities) in honey bees. Results indicated that both neonicotinoids’ sublethal concentrations promote oxidative stress in honey bees and affect their immune defense. NEOCs impair detoxification abilities through reduced glutathione S-transferase activity, as well as disturb immune response due to decreased prophenoloxidase and phenoloxidase activities, which can explain the high toxicity of these substances to honey bees. [ABSTRACT FROM AUTHOR]

Ageing is a complex process common to all living organisms, influenced by different environmental and genetic factors which are difficult to understand. Epigenetic modifications such as DNA methylation, histone posttranslational modification and non-coding RNA affect ageing. Eusocial insects provide an ideal platform for analysing the impact of epigenetic changes on ageing due to their phenotypic plasticity. This study summarizes most of the data published so far on epigenetic changes during ageing in eusocial insects. [ABSTRACT FROM AUTHOR]

Background: Single-walled carbon nanotubes (SWCNTs) have been reported to induce cytotoxicity in different cell lines. Although the mechanisms underlying cytotoxicity are not fully understood, accumulation of reactive oxygen species (ROS) and oxidative damage is considered to be a likely contributing factor. Methods: Human lung carcinoma cells, A549, and human fetal lung fibroblasts, MRC-5 were used to assess the cytotoxicity of SWCNT in the presence and absence of a redox status regulator, N-acetylcysteine (NAC), via the MTT assay. Results: SWCNT induced a nearly three-fold greater loss of viability in A594 vs. MRC-5 cells at ≤250 μg/ml. SWCNT cytotoxicity at higher concentrations was similar for both cell lines, while NAC alone was non-toxic. The cytotoxicity of SWCNT (250 μg/ml) in combination with NAC to A549 cells was significantly decreased at the lowest NAC concentration (1.5 µg/ml), and was similar to NAC treatment alone at that concentration. Higher concentrations of NAC in combination with SWCNT (250 μg/ml) resulted in increased cytotoxicity in both A549 and MRC-5 cells. Conclusion: A549 malignant lung cells are more susceptible to low concentrations of SWCNT vs. normal lung cells, and low concentrations of N-acetylcysteine appear to be cytoprotective, possibly due to its antioxidant properties. [ABSTRACT FROM AUTHOR]

In this study the effect of long term exposure to cadmium (Cd) on Ostrinia nubilalis larval growth, development, survival rate and oxidative status was analyzed. Newly hatched first instar - L1 larvae were reared on a Cd contaminated diet until the larvae reached the final, fifth instar - L5 or developed into pupae. In total, six experimental groups, five treatments (concentrations of Cd in fresh diet: Cd I: 0.73, Cd II: 3.70, Cd III: 6.85, Cd IV: 41.71 and Cd V: 77.53 mg kg−1) and a control group (C) were set up. The results of the experiment showed that exposure to higher concentrations of Cd (41.71 and 77.53 mg kg−1) had a significant influence on development and redox status of O. nubilalis larvae: (1) the development rate was strongly reduced resulting in a prolonged pupation time; (2) the survival rate of larvae was prominently lower; (3) bioaccumulation factor (measured in pupae) was reduced which indicated that larvae could accumulate Cd to a certain level; (4) the level of the lipid peroxidation was significantly higher, which points to oxidative damage; (5) the expression of Mtn was significantly up-regulated while Cat and GPx genes down-regulated. In conclusion, long term exposure to dietary Cd in a concentration of 41.7 mg kg−1 and higher, induced oxidative stress and slowed down growth and development of O. nubilalis larvae. Image 1056 • O. nubilalis larvae accumulates Cd from diet during long term exposure. • Cd in diet (≥ 41.71 mg/kg) alters O. nubilalis larvae growth and development. • Cd in diet (41.71 mg/kg) increases MDA level in O. nubilalis larvae. • Cd in diet (0.73; 6.85; 41.71 mg/kg) up-regulates Mtn gene expression. • Cd in diet (≥6.85 mg/kg) down-regulates Cat and GPx gene expression. [ABSTRACT FROM AUTHOR]

The European corn borer, Ostrinia nubilalis Hbn., is a pest Lepidopteran species whose larvae overwinter by entering diapause, gradually becoming cold-hardy. To investigate metabolic changes during cold hardening, activities of four metabolic enzymes – citrate synthase (CS), lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured in whole-body homogenates of pupae, non-diapausing and diapausing larvae acclimated to 5 °C, −3 °C and −16 °C. The highest CS activity was detected in non-diapausing larvae, reflecting active development, while the highest in vitro LDH activity was recorded in diapausing larvae at temperatures close to 0 °C, evidencing a metabolic switch towards anaerobic metabolism. However, in-gel LDH activity showed that production of pyruvate from lactate is triggered by sub-zero temperatures. The activities of both aminotransferases were highest in non-diapausing larvae. Our findings suggest that during diapause and cold hardening the aminotransferases catalyse production of L-alanine, an important cryoprotectant, and L-aspartate, which is closely tied to both transamination reactions and Krebs cycle. The results of this study indicate that, during diapause, the activity of metabolic enzymes is synchronized with exogenous factors, such as temperatures close to 0 °C. These findings support the notion that diapause is metabolically plastic and vibrant, rather than simply a passive, resting state. [ABSTRACT FROM AUTHOR]

Polyamines are small ubiquitous molecules that have various implications in health and disease. Since the diet is an important source of polyamines, the aim of this study was to determine the content of major natural polyamines (putrescine, spermidine, and spermine) in honey bee products used in human nutrition whose botanical origin was previously determined. The total content of polyamines was the highest in bee pollen (125–712 mg/kg), slightly lower in bee bread (288–602 mg/kg), much lower in royal jelly (68.9–121 mg/kg) and negligibly small in honey samples (2.80–10.8 mg/kg). Spermidine predominated in bee pollen, bee bread and honey, and putrescine in royal jelly. Botanical origin did not affect the total content or the proportion of individual polyamines, which were always representative of the bee product. This study shows that a significant content of polyamines in bee products could contribute to their high nutritional value. [Display omitted] • Data represents the content of total and individual polyamines in honey bee products. • Total polyamine content highlighted bee pollen and bee bread as valuable foods. • Spermidine dominates bee pollen, bee bread, honey; putrescine dominates royal jelly. [ABSTRACT FROM AUTHOR]

The spread and occurrence of the oak lace bug Corythucha arcuata out of its natural distribution area across European and Asian countries has been reported during the past decades. The ecological and economic significance of oak stands and the vulnerability of plants to various abiotic and/or biotic factors requires in-depth knowledge of plant-pest interaction. The present study examined the influence of C. arcuata feeding on the photosynthetic characteristics and gas-exchange parameters, mineral nutrient concentrations and defense mechanisms (the activities of some antioxidant enzymes) of leaves of pedunculate oak. The rate of photosynthesis, transpiration and stomatal conductance were lowered by 58.84, 21.66 and 35.71%, respectively, in comparison to non-infested plants. The concentrations of photosynthetic pigments and activities of antioxidant enzymes, catalase and ascorbate peroxidase, were affected by the presence of C. arcuata. To our knowledge this is the first paper providing a report on the physiological responses of Quercus robur plants exposed to C. arcuata infestation. Understanding the impact of pests, such as the invasive species C. arcuata on physiological processes and vitality of young plants and plant responses, could provide a foundation for efficient preservation of oak forests endangered by the oak lace bug. [ABSTRACT FROM AUTHOR]

Larvae of the European corn borer ( Ostrinia nubilalis Hubn.) were cold acclimated during different phases of diapause to determine if changes in the fatty acid composition lipids occur as part of a programmed diapause strategy, or as a response to low temperatures during winter. Cold acclimation of fifth instar larvae of O. nubilalis during diapause had modest effects further on the readjustments in fatty acid composition of triacylglycerols and phospholipids. Overall, FA unsaturation (UFAs/SFAs ratio) was stable, with the exception of the triacylglycerols fraction after exposure to −3 and −10 °C in mid-diapause (MD) when it significantly increased. Differential scanning calorimetry (DSC) was used to examine phase transitions of total body lipid of cold-acclimated larvae in diapause. Thermal analysis indicated that changes in the melt transition temperatures of whole body total lipids were subtle, but consistent with the modest changes in the level of FA unsaturation observed. We conclude that lipid rearrangements are a function of the endogenous 'diapause program' rather than a direct effect of low temperatures, which proved to have limited impact on lipid changes in diapausing larvae of O. nubilalis. [ABSTRACT FROM AUTHOR]

Abstract: The recent decline in managed honey bee populations, Apis mellifera L. (Hymenoptera: Apidae), has caused scientific, ecological, and economic concern. Research into the formation of reactive oxygen species (ROS), antioxidative defense mechanisms, and oxidative stress can contribute to our understanding of bee survival and conservation of this species. Activities of superoxide dismutase (SOD), catalase (CAT), and glutathione S‐transferase (GST) enzymes together with levels of malondialdehyde (MDA) were measured in summer and winter honey bees sampled from three colonies. One colony was stationary (C1), entering the winter period having accumulated Robinia pseudoacacia L. (Fabaceae) honey, and two were migratory (C2 and C3), entering the winter period with mainly Tilia (Malvaceae) and Brassica (Brassicaceae) honey, respectively. Compared to summer workers, winter worker bees had decreased SOD and GST activity, and MDA level, whereas CAT activity increased in all three colonies. We also demonstrated that seasonality is the main factor responsible for changes in antioxidant enzymes and MDA levels in worker honey bees. Overall, our results indicate a difference between summer and winter worker bees, pointing at a reduced level of antioxidant enzyme defenses during overwintering which may be due to a decrease in production of ROS. The decreased levels of MDA measured in winter honey bees confirm this. As ROS are actively used by insects as a defense mechanism to fight pathogens, we suggest that reduced production of ROS contributes to higher susceptibility of winter honey bees to infections and reduced overwinter survival. [ABSTRACT FROM AUTHOR]

Seasonal changes in the FA composition of triacylglycerols and phospholipids prepared from the whole bodies of non-diapausing and diapausing fifth instar larvae of Ostrinia nubilalis , Hubn. (Lepidoptera: Crambidae) were determined to evaluate the role of these lipids in diapause. Substantial changes in the FA composition of triacylglycerols and phospholipids were triggered by diapause development. This led to a significant increase in the overall FA unsaturation (UFAs/SFAs ratio), attributable to an increase in the relative proportion of MUFAs and the concomitant decrease in PUFAs and SFAs. In triacylglycerols, the significant changes in the FAs composition are the result of an increase in the relative proportions of MUFAs, palmitoleic acid (16:1n-7) and oleic acid (18:1n-9), and a concomitant reduction in the composition of SFAs and PUFAs, mainly palmitic acid (16:0) and linoleic acid (18:2n-6), respectively. Changes in the composition of phospholipids were more subtle with FAs contributing to the overall increase of FA unsaturation. Differential scanning calorimetry (DSC) analysis revealed that the melt transition temperatures of total lipids prepared from whole larvae, primarily attributable to the triacylglycerol component, were significantly lower during the time course of diapause compared with non-diapause. These observations were correlated to the FA composition of triacylglycerols, most likely enabling them to remain functional during colder winter conditions. We conclude that O. nubilalis undergoes remodeling of FA profiles of both energy storage triacylglycerols and membrane phospholipids as an element of its overwintering physiology which may improve the ability to cold harden during diapause. [ABSTRACT FROM AUTHOR]

Abstract: This study compares the composition and biophysical properties of lipids in non-diapausing and diapausing fifth instar larvae of Ostrinia nubilalis Hubn. (Lepidoptera: Crambidae). The majority of fat body lipids in both of these physiological states were comprised of ~90% triacylglycerols (TAGs), whereas the haemolymph contained a more even distribution of all lipid classes. The fatty acid composition and biophysical properties of the fat body lipids differed markedly between non-diapausing and diapausing larvae. Diapause was associated with a dramatic increase in the proportions of palmitoleic acid (16:1n-7) and oleic acid (18:1n-9), with concurrent reductions in palmitic acid (16:0) and linoleic acid (18:2n-6). The increase in the level of unsaturation of the fat body lipids, which caused a marked shift in their phase transitions to lower temperatures, was triggered by diapause rather than low temperatures. Adjustments of fatty acid compositions are likely to be an important component of winter diapause mechanisms, possibly maintaining the fluidity of cell membranes and the functionality of the organism during lower winter temperatures. [Copyright &y& Elsevier]

Cold hardiness in the Arctic Collembola Megaphorura arctica (Tullberg), formerly Onychiurus arcticus, has been the subject of extensive studies over the last decade. This species employs an unusual strategy known as cryoprotective dehydration to survive winter temperatures as low as −25 °C. To expand knowledge of cryoprotective dehydration in M. arctica, the present study investigates how a reduction in ambient temperature affects the fatty acid composition of the total body lipid content along with polar (mainly membrane phospholipids) and nonpolar (mainly triacylglycerols) lipids. Most ectothermic animals compensate for changes in fluidity by regulating fatty acid composition, a process often described as homeoviscous adaptation. In M. arctica, changes in the fatty acid composition of total body lipid content during cold treatment are only moderate, with no clear pattern emerging. However, the levels of unsaturated fatty acids in the polar lipids increase with cold exposure, largely attributable to 16 : 1( n− 7), 18 : 1( n− 9), 18 : 3( n− 6) and 18 : 3( n− 3), whereas unsaturated fatty acid levels in the nonpolar lipids correspondingly decrease. These results suggest a reallocation of fatty acids between the two lipid pools as a response to a temperature reduction of 6 °C. Because of hypometabolism, a characteristic of cold adaptation, such a mechanism could be less energy demanding than de novo synthesis of fatty acids and may comprise part of an adaptive homeostatic response. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) is a non-essential metal that is highly toxic to all living forms, characterized by an extremely high affinity for thiol (SH) groups. The aim of this work was to identify and experimentally verify metallothionein gene and to analyze the role of biological thiols in stress induced by short-term Cd exposure in Ostrinia nubilalis , one of the most important corn pests. The coding region of a metallothionein (MT) gene in O. nubilalis was identified, encoding protein, OnMT1, which contains 46 amino acids, including 12 cysteine residues, and has no aromatic amino acids. Phylogenetic analysis revealed that OnMT1 clustered together with metallothionein from Bombyx mori. Structural bioinformatics analysis strongly suggests that OnMT1 is a metallothionein with affinity for multiple transition metals. Further, in order to elucidate the role of biological thiols, O. nubilalis L5 larvae were exposed to increasing Cd concentrations in diet (6.85, 41.71, 77.35 mg kg-1) during a 48 h period, after which Cd concentration in larvae was measured (3.50, 12.02, 47.37 mg kg-1, respectively). Due to short-term Cd exposure, concentration of free protein SH groups and relative expression of OnMT1 and thioredoxin (Trx) genes was elevated, while the reduced glutathione content remained unchanged. The presented results provide evidence that OnMT1 plays a role in Cd detoxification and homeostasis, and confirm the importance of biological thiols, especially OnMT1 and Trx, in the early response of O. nubilalis to Cd poisoning, indicating interaction between Cd and thiol-linked redox reactions. Insects provide valuable insight into molecular adaptations to metals. [Display omitted] • Little is known about the effects of metals on insects at the molecular level. • A metallothionein gene encoding OnMT1 protein was identified in Ostrinia nubilalis. • There is dose response between Cd concentrations in larvae and diet, after 48 h. • The role of thiols, especially OnMT1 and Trx, in early response to Cd is shown. • Insects provide valuable insight into molecular adaptations to metals. [ABSTRACT FROM AUTHOR]

Honeybee colony losses have been a focus of research in the last years, due to the importance of managed honeybee colonies for economy and ecology. Different unfavorable conditions from the outside environment have a strong impact on the hive health. The majority of losses occur mainly during winter and the exact reason is not completely understood. Only a small number of studies are dealing with content of bioelements, their function and influence on honeybee physiology. The aim of the present study was to determine seasonal and spatial variations in content of bioelements and non-essential elements, in hemolymph and whole body of honeybees originating from three regions with different degrees of urbanization and industrialization. Concentrations of 16 elements were compared: macroelements (Ca, K, Mg, Na), microelements (Cu, Fe, Mn, Zn) and non-essential elements (Al, Ba, Cd, Co, Cr, Ni, Pb, Sr) in samples collected from 3 different environments: Golija (rural region), Belgrade (urban region) and Zajača (industrial region). Content of bioelements and non-essential elements in honeybees was under noticeable influence of the surrounding environment, season and degree of honeybee activity. Hemolymph was proven to be helpful in differentiating air pollution from other sources of honeybee exposure. The results of our study demonstrated that bees can be successfully used as biomonitors since we have observed statistically significant differences among observed locations, but unless compared locations are exposed to excessively different pollution pressures, it is essential that all bees should be collected at the same season. Unlabelled Image • Environment and season influence the content of measured elements in honeybees. • To be used for biomonitoring, bees should be collected during the same season. • Hemolymph provided better understanding of seasonal dynamic of elements content. • In highly contaminated areas, dominant way of exposure is PM deposition on bee body. [ABSTRACT FROM AUTHOR]