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Varroa destructor is a parasitic mite that attacks the honey bee, and previous studies have suggested that parasitosis caused by this mite is accompanied by immunosuppresion in the host. In this study, the effect of mite infestation on the expression of the lysozyme-1 (lys-1) gene and lysozyme activity in Apis mellifera carnica was determined. The experiment was carried out on the five developmental stages of honey bee workers and drones. Developmental and gender-related differences in gene expression and lysozyme activity were observed in a Varroa destructor-infested brood. The relative expression of the lys-1 gene increased in a infested worker brood and decreased in a drone brood except for P3 pupae. In the final stage of development, the lys-1 gene expression was significantly lower in infested newly emerged workers and drones. Changes in the relative expression of the lys-1 gene in infested individuals was poorly manifested at the level of enzyme activity, whereas at the two final stages of development (P5 and I) there was a positive correlation between relative lys-1 expression and lysozyme activity in infested bees of both genders (r=0.988, r=0.999, respectively). The results of this study indicate that V. destructor influences the lysozyme-linked immune response in bees.

Plants adapt to extremely low temperatures in polar regions by maximizing their photosynthetic efficiency and accumulating cryoprotective and osmoprotective compounds. Flowering plants of the family Poaceae growing in the Arctic and in the Antarctic were investigated. Their responses to cold stress were analyzed under laboratory conditions. Samples were collected after 24 h and 48 h of cold treatment. Quantitative and qualitative changes of sugars are found among different species, but they can differ within a genus of the family Poaceae. The values of the investigated parameters in Poa annua differed considerably depending to the biogeographic origin of plants. At the beginning of the experiment, Antarctic plants were acclimatized in greenhouse characterized by significantly higher content of sugars, including storage reserves, sucrose and starch, but lower total protein content. After 24 h of exposure to cold stress, much smaller changes in the examined parameters were noted in Antarctic plants than in locally grown specimens. Total sugar content and sucrose, starch and glucose levels were nearly constant in P. annua, but they varied significantly. Those changes are responsible for the high adaptability of P. annua to survive and develop in highly unsupportive environments and colonize new regions.

Varroa destructor is a parasitic mite of the Western honey bee. The activity of five antioxidant enzymes of V. destructor were analysed. Glutathione content and total antioxidant status was also evaluated. Our results suggest that antioxidant enzymes constitute the main line of defense against ROS in V. destructor, whereas low-molecular-weight antioxidants play a limited role in the antioxidant system of mites.

The pathogenesis of varroasis has not been fully explained despite intensive research. Earlier studies suggested that parasitic infections caused by Varroa destructor mites were accompanied by immunosuppression in the host organism. The objective of this study was to analyse the influence of varroasis on one of the immune pathway in Apis mellifera measured by the expression of the prophenoloxidase (proPO) gene and the enzymatic activity of this gene’s product, phenoloxidase (EC 1.14.18.1). An evaluation was done of five developmental stages of honey bee workers and drones. The relative expression of proPO decreased in infected individuals. The only exceptions were worker prepupae (PP) and drone pupae with brown eyes and dark brown thorax (P5) where propo gene expression was 1.8-fold and 1.5-fold higher, respectively, than in the control. Phenoloxidase (PO) activity was 2.8-fold higher in infected pp workers and 2-fold higher in p5 drones in comparison with uninfected bees. Phenoloxidase activity was reduced in the remaining developmental stages of infected workers and drones. The relative expression of proPO was positively correlated with the relative PO activity in both workers (r = 0.988) and drones (r = 0.996). The results of the study indicate that V. destructor significantly influences the phenoloxidase-dependent immune pathway in honey bees.

The colony collapse disorder is a growing problem world-wide. For this reason, we were prompted to search for natural and harmless agents that could improve the living conditions of honey bees. This group of agents includes exogenous antioxidants, such as ascorbic acid, which boost natural immunity. We analysed the effect of vitamin C supplementation on carbohydrate metabolism in the developing honey bee worker brood. The total carbohydrate content and the concentrations of glycogen, trehalose, maltotriose, fructose, and glucose were estimated. The correlations between sugar content and the activity of the main hydrolases of carbohydrate metabolism - α-amylase, glucoamylase, trehalase, maltase, and sucrase - were determined. The addition of vitamin C to the diet of wintering bees did not impair their sugar metabolism. Vitamin C supplements exerted a positive effect by significantly increasing glycogen and trehalose concentrations in the initial phase of development and in newly emerged workers. Vitamin C did not induce significant changes in the developmental profile of carbohydrate degrading enzymes, except for the earliest stage of larval development when enzyme activity levels were below those noted in the control group.

Osmia rufa is a solitary bee that is used commercially for pollinating crops. The bee enters obligatory diapause as an imago. The activity of proteolytic enzymes during diapause has not been investigated. We studied the proteinase activity on four substrates - casein, haemoglobin, bovine serum albumin (BSA ), and gelatine - during diapause (from October to March) and in newly hatched males and females in April. During diapause, greater fluctuations in enzyme activity levels were noted in males than in females, and a significant decrease in male enzyme activity was observed in January and March. Male enzymes were most effective in decomposing gelatine; whereas, female enzymes were equally effective in hydrolysing gelatine and BSA . The differences in substrate preferences between male and female enzymes were particularly pronounced in October and in the newly hatched individuals. The levels of gelatinolytic activity likely indicate that a high proportion of proteinases in O. rufa are elastase-like enzymes. They are involved in the digestion and remodelling of proteins with numerous peptide bonds formed by amino acids with short side-chains.

The aim of the present research was to determine the basic composition of the organic compounds present in extracts from Varroa destructor, a parasitic mite of the honeybee. The total protein content was 9.16 ± 0.82 mg/100 mg of body weight, lipid content was 9.81 ± 1.99 mg/100 mg, and carbohydrate content was 26.67 ± 4.52 mg/100 mg. The triacylglycerol content was 2.40 ± 0.86 mg/100 mg and the cholesterol content was 0.14 ± 0.02 mg/100 mg. Thin layer chromatography indicated that phospholipids comprised the major part of the lipid component; cephalins (78%), cerebrosides (16%), and lecithins (6%) were identified in the phospholipid pool. Glucose (23.6 ± 4.52 mg/100 mg) was the main carbohydrate, followed by glycogen (5.43 ± 1.23 mg/100 mg) and trehalose (0.35 ± 0.07 mg/100 mg). Enzyme-linked immunosorbent assays detected two major glycogen metabolism enzymes, glycogen phosphorylase and glycogen synthase. Among the enzymes metabolising disaccharides, maltase (24.7 ± 2.38 μmol/mg protein) and trehalase (14.81 ± 5.21 μmol/mg protein) presented the highest activity. Saccharose and lactose were hydrolysed to a minor extent. These are the first measurements of the basic composition of the mite body. Although these data are not exhaustive, they may serve as the basis for further research on the metabolism of V. destructor, particularly concerning lipid metabolism.

The red mason bee (Osmia bicornis) is a highly effective pollinator that is exposed to various xenobiotics. The organism's potential resistance to the toxic effects of xenobiotics can be determined based on cholinesterase activity. The activity of cholinesterases (ChEs) towards acetylcholine (ACh) and butyrylcholine (BCh) was determined in extracts of diapausing (between October and late March) and flying bees (May). In both males and females, enzyme activity was higher towards ACh than towards BCh. The ratio of ACh/BCh activity was determined in the range of 1.43 to 4.15 in diapausing females and 3.00 to 7.18 in diapausing males. No significant changes in ChE activity towards ACh were observed in females before December and in males before February. Enzyme activity towards ACh increased dynamically in the second half of March. Enzyme activity towards BCh remained stable in both sexes until mid-March, after which it increased significantly. Excluding mid-March, enzyme BCh activity was significantly higher in females than in males. The activity of carboxylesterase towards 4-p-nitrophenyl butyrate was determined in females to assess the involvement of non-specific esterases in the hydrolysis of choline esters. Carboxylesterase activity was low in comparison with cholinesterase activity, and it remained practically unchanged throughout diapause, suggesting that choline esters in female O. bicornis extracts were hydrolyzed mainly by acetylcholinesterases.

Cancer diseases are an important problem of the contemporary clinical medicine and pharmacology. Chemotherapy using cytostatic drugs has developed in the last 30 years. At present, it is the main method of therapy of metastatic cancers. Cytostatics have an important role in the cancer therapy. They have particular meaning in the therapy of solid and hematological tumors. However, using cytostatic drugs is limited due to their toxic effects on healthy cells. In last years, the decrease of toxicity of cytostatic drugs and the increase of their therapeutic properties are intensively investigated. In this paper, the polymer carriers of cytostatic drugs (mainly epirubicin and cyclophosphamide) and examples of their applications were described.

Bees of the genus Osmia are being used in crop pollination at an increasing rate. However, a short life expectancy of adult individuals limits the feasibility of their use. Cocoons of the red mason bee, Osmia rufa L. (Hymenoptera: Megachilidae), can be stored at 4° C in a postdiapause state, and adult bees can be used for pollination outside their natural flight period. The period of storage in this form has an unfavorable influence on the survival rate, life expectancy, and fertility of the bee. It was suggested that the negative results are connected with exhaustion of energy reserves. To test this hypothesis, the present study examined the contents of protein, carbohydrates, lipids, and the activities of some enzymes, and their degradation in red mason bees that emerged in spring according to their biological clock and in summer after elongated diapause. It was found that postdiapause artificially elongated by 3 months caused significant decreases in body weight, total sugar, glycogen, lipids, and protein content in O. rufa. Glucose level was highest in bees that emerged in the summer, which was coincident with increased activities of maltase and trehalase. The activities of sucrase and cellobiase were not changed, while amylase activity was considerably decreased. The activities of triacylglycerols lipase and C2, C4, C10 carboxylesterases were highest in bees that emerged in July. Low temperatures restrict O. rufa emergence, and during prolonged postdiapause, metabolic processes lead to significant reductions of structural and energetic compounds.

New polymeric conjugates were prepared coupling ofloxacin to two-, three-, four and six-arm, star-shaped poly(ɛ-caprolactone) and polylactide. The homopolymers were synthesized via ring-opening polymerization of ɛ-caprolactone, l-lactide and rac-lactide in the presence of glycerol, penthaerythritol, dipentaerythritol and poly(ethylene glycol) as initiators and stannous octoate as a catalyst. The conjugates were characterized by GPC, MALDI-TOF MS, NMR, IR and viscosity methods. Content of Sn has been investigated in polymers by electrothermal atomic absorption spectrometry. Toxicity of monomers, initiators and polymers were evaluated with bacterial luminescence test and two protozoan assays. The in vitro release of ofloxacin from obtained conjugates in pH 7 was investigated., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Extracts of Anisakis simplex third (L3) and fourth (L4) larval stages were assayed for protein content and activity and properties of alpha-amylase, glucoamylase and glycogen phosphorylase. Protein content in L4 was twice that in L3. SDS-PAGE applied to both larval stages revealed 22 protein fractions in each, including five stage-specific fractions in each larval stage. The L3 extracts contained three amylase isoenzymes: alpha 1, alpha 2 and alpha 3; their molecular weights were 64, 29 and 21 kDa, respectively. Only one amylase isoenzyme (64 kDa) was found in the L4 extracts. Glycogen in L3 was found to be broken down mostly by hydrolysis because of low glycogen phosphorylase activity. The alpha-amylase activity in L4 was higher than that in L3 by half and the glycogen phosphorylase activity was ten times higher. In addition, the same enzymes isolated from L3 and L4 were found to differ in their properties. These differences could be manifestations of metabolic adaptations of A. simplex larvae to host switch from fish (L3) to mammals (L4), i.e. adaptations to a new habitat.

Contracaecum rudolphii is the parasitic nematode of fish-eating birds. In the extracts from female, male and larvae L3 and L4 isolated from the alimentary tracts of black cormorants the activity of five antioxidant enzymes: superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione transferase (GST), glutathione reductase (GR), catalase (CAT) and the content of ascorbate and total antioxidative status (TAS) were determined. They can be put in order according to the activity growth: GPX, SOD, GST, CAT and GR. The activity of GPX were very low in the nematodes' extracts (1.23-7.67 microU/mg). CAT had higher activity (0.47-0.72 U/mg). The activity of GR was the highest (50.51-69.88 U/mg). SOD activity in the female was higher by ca. 50% than in the male while GST activity was at similar levels. GR and CAT activities were higher by ca. 30% in the male than in the female nematodes. GST and GPX activity and TAS in larvae L3 were significantly lower than in the adult nematodes or in L4 larvae. The activity of GPX, GR and CAT was lower in L4 larvae than in the adult male (p<0.05). The content of ascorbate was almost the same in all stages of parasite development (0.21-0.38 mg/g). The above results indicate differences in antioxidant systems related to both the sex and the developmental stage of C. rudolphii.

Background: Enzymatic activity is an indicator of an organism's metabolic rate which depends on, i.e., environmental conditions, developmental stage, physiological stage, and sex. The API ZYM test was applied to compare activities of 19 hydrolases of female and male Hysterothylacium aduncum., Material and Methods: Sexually mature nematodes were isolated from eelpout individuals caught in the Gulf of Gdańsk. Enzymatic activity of the hydrolases and the protein content was determined in nematode extracts using API ZYM and Bradford's method, respectively., Results: The females and males tested showed a total of 13 enzymes to be active. The males showed additionally the presence of alpha-fucosidase. Acidic and alkaline phosphatases had very high activities in both sexes; short-chain fatty acid esterases, leucine and valine aminopeptidases, alpha-glucosidase, and N-acetylglucosaminidase were highly active. H. aduncum showed no trypsin- and chymotrypsin-specific activities; similarly, no activity of alpha-galactosidase, alpha-mannosidase, and beta-glucuronidase was revealed. Except for lipase (C14), hydrolases were more active in females than in males, which is related to metabolic rate being higher in females due to their reproductive function., Conclusion: Comparison of the results obtained with earlier data produced with API ZYM allowed suggesting that the hydrolase pattern may be more affected by habitat in the host than by the taxonomic affiliation of nematode.

Introduction: The influence of infection with two species of entomopathogenic nematodes of Steinernematidae family on metabolism of glycogen and trehalose of the host was studied., Material and Methods: Last instar larvae (L7) of Galleria mellonella were experimentally infected with Steinernema affinis and S. feltiae. At 6, 12, 18 and 24 h after infection concentrations of trehalose and glycogen as well as activity of trehalase and alpha-amylase were determined., Results: The content of glycogen was lower in insects infected with S. feltiae than in the controls and animals infected with S. affinis. The content of trehalose was higher in insects from both infected groups than in the controls. Its concentration was slightly higher in larvae infected with S. affinis than in those infected with S. feltiae. The activity of alpha-amylase after infection with S. affinis was low. It was significantly higher in insects infected with S. feltiae. In animals of both infected groups, following a significant reduction at 6 h, the activity of trehalase remained at a similar level, higher than in the controls. In the paper the effects of infection with (i) different species of entomopathogenic nematodes and (ii) the importance of the developmental stage of the insect-host for changes in its metabolism of glycogen and trehalose were discussed.

Background: The influence of infection with two species of entomopathogenic nematodes of Steinernematidae family on the activity of superoxide dismutase (SOD) of the host was studied., Material and Methods: Last instar larvae of Galleria mellonella were experimentally infected with Steinernema affinis and S. feltiae at 20 invasive juveniles per insect. At 6, 12, 18, 24 and 36 h after infection activity of SOD was determined in extracts from infected and control insects., Results: The activity of SOD decreased gradually in the controls during the experiment. The activity of enzyme was 2-4-times higher in insects from both infected groups than in the control. During the first 12 h of infection the activity of SOD in insects infected with S. feltiae was higher than in those infected with S. affinis, then the activity of enzyme in the insects of both infected groups stayed at a similar level. A significant decrease of SOD activity in infected was recorded in second day of the infection.

The proteins level and activities of acid and alkaline proteases in whole body extracts of drone prepupae of Apis mellifera naturally infested with Varroa destructor were studied. The infested and a non-infested group did not differ significantly in their total protein content. However, some differences in protein profiles were found. A lack of three protein fractions of moderate and lower molecular weight in infested prepupae was noted. Moreover, some differences in the quantity of protein in most of the fractions were observed. The activity of acid proteases from infested prepupae was lower (p < 0.05) compared with the activity of these proteases from the non-infested one group. The infested drone had higher activity of alkaline proteases than non-infested but this difference was not statisticaly significant.

Activity of hydrolases during the third and fourth larval stage of Anisakis simplex was identified by applying the API ZYM test method. In A. simplex larvae the activity of phosphatases was high, particularly that of acid phosphatase (40 nmol/mg(-1)). Among esterases lack of activity of lipase (C14) is worth noticing while the activity of esterases (C4) and (C8) was high. The activity of those later two enzymes was higher in L3 larvae than in L4 larvae. The highest activity in the subclass of glucosidases was recorded for beta-fucosidase and N-acetyl-beta-glucosaminidase. A higher activity in L3 larvae than in L4 larvae was recorded for: beta-glucuronidase and N-acetyl-beta-glucosaminidase (2-fold) and beta-fucosidase (3-fold). Differently the activity of beta-galactosidase and beta-glucosidase was higher in L4 larvae than in L3 larvae. The tests did not show activity of alpha-galactosidase, beta-glucosidase and alpha-mannosidase on both larval forms.

Concentration of glycogen, maltose, and trehalose as well as the activity of hydrolases catabolising them: alpha-amylases, glucoamylase, maltase, and trehalase were studied during a 48 h infection of Galleria mellonella (Lepidoptera) instar larvae infected with infective Heterorhabditis zealandica (20 nematodes/insect). The content of trehalose in the infected insects during the first day of infection was higher (p < 0.05) than in the controls. The concentration of maltose and glycogen were similar in both groups despite the fact that the activity of all examined alpha-glycosidases was generally much higher in the infected insects than in the controls.

Acid trehalase (EC 3.2.1.28) was isolated from muscle of Ascaris suum by fractionating with ammonium sulfate, acetone and column chromatography on DEAE-cellulose and phenyl sepharose CL-4B. The purified homogeneous preparation of native acid trehalase exhibited a molecular mass of 76 kDa and of 38 kDa on SDS-PAGE. The enzyme has the optimum pH 4.9, pI 4.3, Km of 6.6 mM and Vmax=34.5 nM min(-1) x mg(-1). Besides trehalose, it hydrolyses sucrose, isomaltose and maltose and, to a lesser degree melezitose, and it does not act on cellobiose and lactose. Acid trehalase was activated by MgCl2, KNO3, NaCl, CaCl2, CH2ICOOH and p-chloromercuribenzoate and inhibited by EDTA, ZnSO4 and FeCl3.

Similar patterns of hydrolases were observed in three species representing two genera of entomopathogenic nematodes: Steinernema affinis, S. feltiae, and Heterorhabditis zealandica. The same enzymes were present in the studied nematodes but they differed in the level of activity of individual sub-classes of enzymes. A higher activity of esterases and proteolytic enzymes could be observed for H. zealandica than for S. affinis and S. feltiae. On the other hand, the activity of glycosidases in steinernematids was generally much higher than in H. zealandica.

The activity of alpha-amylase, glucoamylase, maltase, trehalase, glycogen phosphorylase and trehalose phosphorylase was measured in extracts from larval and adult Hysterothylacium aduncum (Rudolphi, 1802), parasitic nematode of marine fish. The content of glycogen and trehalose in the worm's body was also determined. Both the hydrolytic and phosphorolytic paths of sugar decomposition are present in H. aduncum. In the larvae glycogen was utilised mainly via the hydrolytic path. In the adults the activities of phosphorolytic enzymes were higher than in the larvae. In both stages the activity of trehalose phosphorylase is present. In adult nematodes it is uncommonly high. The dominating sugars in the adults were glucose and glycogen, while in the larvae it was trehalose.

The content of glycogen, glucose and trehalose was measured in larvae and adults of Cystidicola farionis, the parasite isolated from the swim bladder of Osmerus eperlanus from Vistula Lagoon. Activity of glycogen phosphorylase, alpha-amylase, glucoamylase, maltase, trehalase, and trehalose phosphorylase were measured. The highest activity was recorded for alpha-amylase 10.07 +/- 0.97 mu/mg and 7.47 +/- 0.24 mu/mg, next maltase 1.34 +/- 0.63 micromol/mg and 2.06 +/- 1.65 micronol/mg respectively for larvae and adults. The activity of glucoamylase was nearly the same for adults and larvae (about 0.20 micromol/mg). The trehalase activity was higher at adults (0.49 +/- 0.42 micromol/mg) than at larvae (0.18 +/- 0.12 micromol/mg). The activity of glycogen phosphorylase was much higher at larvae (3.58 +/- 1.49 micromol/mg) than at adults parasite (0.10 +/- 0.02 micromol/mg). The trehalose phosphorylase was present in both stages of parasite, but its activity was low. The content of glycogen and glucose was two-times higher in the adults' body than in larvae.

The objective of this work was to examine the influence of heavy metals ions, zinc and lead, in external environment on the development of Ascaris suum to invasive stage. The culture mediums contained 25.5, and 0.5 mM Pb(NO3)2 or ZnSO4. After 6 weeks of development in 26 degrees C in the control 75.17% of eggs reached the invasive stage. Ions of lead and zinc, in a way depending on concentration inhibited development of embryos. The invasive stage in the presence of 25.5 and 0.5 mM Pb(NO3)2 were achieved by 37.89% , 44,75% and 64.63% of eggs and in ZnSO4 solutions with the same concentrations of ions 45.40%, 50.36% and 66.61% respectively. Many of eggs in 25 mM solutions of both metals stopped at the blastula stage.

The studies were conducted during the school year of 1996/1997 among the children at the age of 5, 6 and 7 years old from Olsztyn and nearby municipality of Purda. In total, 151 children were examined by Graham's method. The presence of eggs of the parasite was confirmed in 13.3% of the total examined population. The infestation was lower in Olsztyn (12.0%) and the lowest in Purda (9.4%) but in the former state-owned Farm village Prajłowo it reached 27.8%. In general, the pinworms were found more frequently in 7 year old children (23.33%) than in case of the others (8.69% and 7.14% for 5 and 6 year old children respectively). In the study, younger boys were more frequently infested than the girls.

Chickens 10 days old were infected with 500 eggs of Ascaridia galli. They were fed with diet containing 11% or 19% protein. Some of them were given beta-carotene in a dose equivalent to daily requirement of vitamin A, others were given double dose of beta-carotene. The intensity and extensity of invasion, the body weight, and activities of alpha-amylase, lipase and trypsin in pancreas and duodenal content were measured. Application of beta-carotene (especially in a double dose) caused the decrease in the invasion index and moderated the changes in the activity of digestive enzymes, occurring in the infected chickens.

The development of Ascaris suum eggs maintained in the culture containing moxidectin (Cydectin, Cyanamid) in concentration 1, 5, 25, and 50 micrograms/ml was studied. The abnormalities and slowing down of the rate of eggs development, depending on the drug concentration, were observed. Exposure of eggs to moxidectin in concentration 1 and 5 microgram/ml resulted in one- or two-week delay of development. In the higher concentration of moxidectin (25 micrograms/ml) only 18% of eggs became invasive. In the presence of 50 micrograms/ml of the drug only few eggs developed to invasive stage after 7 weeks. The development of embryons was stopped more often in the blastula or gastrula stage. The drug's solvent in the highest concentration restricted the development of Ascaris eggs by about 10%. Unembryonated eggs of the worm appeared to be unaffected by 50 micrograms/ml of moxidectin treatment lasting 1-4 days, but for slowing down a little the development rate. The same concentration of the drug in the culture during the first four days of development resulted in 20% reduction of invasive stage in comparison to the control.