Acute and Chronic Exposure to Linagliptin, a Selective Inhibitor of Dipeptidyl Peptidase-4 (DPP-4), Has an Effect on Dopamine, Serotonin and Noradrenaline Level in the Striatum and Hippocampus of Rats.

Linagliptin is a selective dipeptidyl peptidase-4 (DPP-4) inhibitor that indirectly elevates the glucagon-like peptide-1 (GLP-1) level. The aim of the present study was to check whether linagliptin has an influence on neurotransmission in rat brain. Rats were acutely and chronically exposed to linagliptin (10 and 20 mg/kg, intraperitoneally (i.p.)). Twenty-four hours later, the striatum and hippocampus were selected for further studies. In neurochemical experiments, using high-performance liquid chromatography with electrochemical detection (HPLC-ED), the concentrations of three major neurotransmitters-dopamine, serotonin and noradrenaline-and their metabolites were measured. The analysis of mRNA expression of dopamine (D1 and D2), serotonin (5-HT-1 and 5-HT-2) and noradrenaline (α1 and α2a) receptors was also investigated using real-time quantitative reverse transcription polymerase chain reaction (RQ-PCR) in the same brain areas. Linagliptin has the ability to influence the dopaminergic system. In the striatum, the elevation of dopamine and its metabolites was observed after repeated administration of that linagliptin, and in the hippocampus, a reduction in dopamine metabolism was demonstrated. Acute linagliptin exposure increases the serotonin level in both areas, while after chronic linagliptin administration a tendency for the mRNA expression of serotoninergic receptors (5-HT1A and 5-HT2A) to increase was observed. A single instance of exposure to linagliptin significantly modified the noradrenaline level in the striatum and intensified noradrenaline turnover in the hippocampus. The recognition of the interactions in the brain between DPP-4 inhibitors and neurotransmitters and/or receptors is a crucial step for finding novel discoveries in the pharmacology of DPP-4 inhibitors and raises hope for further applications of DPP-4 inhibitors in clinical practices.

The effect of early administration of antibiotics or feeding a diet containing a coccidiostat on inflammatory responses and the morphological structure of selected organs of the immune system in young meat-type turkeys.

It was assumed that early administration of enrofloxacin or doxycycline may impair immune function and alter the morphology of organs of the immune system in turkeys, and that diets containing the coccidiostat monensin, an ionophore antibiotic, can exert similar effects. The aim of this study was to determine whether early antibiotic administration or feeding a diet containing a coccidiostat affect immune function in young turkeys. The experiment had a completely randomized design, with 8 groups (a total of 3,080 one-day-old turkeys), 7 replicate pens per group and 55 birds per pen. The experiment had a 2-factorial design, with 4 treatments (C-control, M-monensin, E-enrofloxacin, and D-doxycycline) and 2 groups of birds (vaccinated and unvaccinated) per treatment. Control group birds did not receive the coccidiostat or antibiotics. Group M was administered monensin at 90 mg/kg feed for the first 5 d of life, group E received enrofloxacin at 10 mg/kg BW, added to drinking water, for the first 5 d of life, and group D received doxycycline at 50 mg/kg BW, added to drinking water, for the first 5 d of life. One-day old turkeys from groups C+, M+, E+, and D+ were administered live-attenuated vaccines against turkey rhinotracheitis (TRT) (Poulvac TRT; Zoetis, Parsippany, NJ) and Newcastle disease (ND) (Nobilis ND clone 30; Merck, Rahway, NJ) by coarse spray; 28-day-old birds were administered a subcutaneously injected inactivated vaccine against Ornithobacterium rhinotracheale (ORT) (Ornitin, Phibro, Poland). Turkeys from groups C-, M-, E-, and D- were not vaccinated. It was found that early administration of enrofloxacin or doxycycline, or feeding a diet containing monensin, did not weaken the immune system of turkeys. The administration of monensin, in particular when combined with vaccination, was least effective in inhibiting inflammatory responses. Histological changes in immunocompetent organs (fatty degeneration) were also most severe in birds receiving monensin, followed by those administered doxycycline and enrofloxacin. The observed changes were exacerbated by vaccination.

The Effect of Copper Nanoparticles and a Different Source of Dietary Fibre in the Diet on the Integrity of the Small Intestine in the Rat.

The aim of the study was to verify the hypothesis regarding the effect of recommended (6.5 mg/kg) or enhanced (13 mg/kg) level of CuNPs in the diet in combination with different types of dietary fibre-cellulose (control), inulin, pectin or psyllium-on selected biological parameters of intestinal integrity in rats. Rats were randomly divided into 10 groups. The first two groups were fed a control diet that contained cellulose, and a mineral mixture with standard or enhanced content of CuCO3. Experimental groups were fed a diet supplemented with CuNPs (6.5 or 13 mg/kg) and combined with different types of fibre (cellulose, pectin, inulin or psyllium). After the feeding period, blood and small intestine samples were collected for further analysis. Replacing CuCO3 by CuNPs in the diet positively reduced the level of lactic acid and apoptosis markers in the small intestine; however, it also resulted in the intensification of DNA oxidation. The most beneficial effect on DNA repair mechanisms is related to inulin, while pectin has the greatest ability to inhibit inflammatory processes that induce the apoptotic death of cells in the small intestine. Our results suggest that dietary fibre supplementation protects the small intestine against potentially harmful, oxidative effects of CuNPs by intensifying the intestinal barrier.

Effect of Copper Nanoparticles in the Diet of WKY and SHR Rats on the Redox Profile and Histology of the Heart, Liver, Kidney, and Small Intestine.

The aim of this experiment was to test the effect of the partial or complete replacement of traditional CuCO3 in the diet of rats with copper nanoparticles (CuNPs) on the biochemical parameters, redox status, and histomorphometry of their tissues. Normotensive male Wistar-Kyoto rats (WKY) were allocated to three groups. Three analogous groups of spontaneously hypertensive rats (SHR) were also formed. The WKY and SHR rats received copper in a standard daily dose-6.5 mg/kg CuCO3 or CuNPs (100% replacement) or 3.25 mg/kg CuCO3 plus 3.25 mg/kg CuNPs (50% replacement)-for 8 weeks. Next, blood, heart, small intestine, liver, and kidney samples were collected. The activity of alanine aminotransferase, aspartate aminotransferase, creatine kinase, and gamma-glutamyl transferase and the content of creatinine and urea acid were measured in the plasma. The collected tissues were subjected to a histological evaluation, and redox status parameters (catalase and superoxide dismutase activity, malondialdehyde and glutathione content) were determined. The replacement of CuCO3 with CuNPs in the diet may exacerbate the negative changes induced by hypertension in the heart, liver, and intestines. However, it seems that it is only in the case of the liver where the observed changes may be due to an increase in oxidative reactions resulting from the inclusion of CuNPs.

Linagliptin, a Selective Dipeptidyl Peptidase-4 Inhibitor, Reduces Physical and Behavioral Effects of Morphine Withdrawal.

(1) Background: Recent data indicate that receptors for GLP-1 peptide are involved in the activity of the mesolimbic system. Thus, the purpose of the present study was to examine the effect of the selective dipeptidyl peptidase-4 (DPP-4) inhibitor, linagliptin, on morphine dependence in mice. (2) Methods: Morphine dependence in mice was obtained by administration of increasing doses of morphine for eight consecutive days, twice a day. On the 9th day of the experiment, the naloxone-induced (2 mg/kg, ip) morphine withdrawal signs (jumping) were assessed. Moreover, behavioral effects of short-term (60 h after morphine discontinuation) and long-term (14 days after morphine discontinuation) morphine withdrawal were observed. In terms of behavioral effects, the depressive effect in the forced swim test and anxiety in the elevated plus maze test were investigated. Locomotor activity of mice was also studied. (3) Results: The administration of linagliptin (10 and 20 mg/kg, ip) for 8 consecutive days before morphine injections significantly diminished the number of naloxone-induced morphine withdrawal signs (jumping) in mice. In addition, the cessation of morphine administration induced depressive behavior in mice which were observed during short- and long-term morphine withdrawal. Linagliptin administered during morphine withdrawal significantly reduced the depressive behavior in studied mice. Furthermore, the short-term morphine withdrawal evoked anxiety which also was reduced by linagliptin in mice. (4) Conclusions: The present study reveals that GLP-1 receptors are involved in morphine dependence. What is more, linagliptin might be a valuable drug in attenuating the physical symptoms of morphine dependence. It might be also a useful drug in reducing emotional disturbances which may develop during the morphine withdrawal period.

In vitro and in vivo evaluation of antioxidant and neuroprotective properties of antipsychotic D2AAK1.

The susceptibility of neurons to free radical toxicity partially underlies the pathomechanism of neurodegenerative diseases. On the other hand, excitotoxicity also contributes to neurodegeneration. Our previous studies demonstrated the unique properties of D2AAK1 as a potent multi-target ligand of aminergic G protein-coupled receptors (GPCRs) which dose-dependently stimulates growth, survival of neurons, and promotes their integrity. The aim of our study was to investigate the potential neuroprotective and antioxidant properties of D2AAK1. Here we show that D2AAK1 activates cellular and molecular neuroprotective mechanisms, prevents cells from excitotoxicity and free radicals. Furthermore, D2AAK1 induced no genotoxic events in neuronal cells in vitro. Most importantly, D2AAK1 protects neurons from the effects of high temperatures by molecular chaperones activation. The D2AAK1 effects on selected organs was further evaluated in mice and no pathological changes were observed after chronic administration. In the light of our experiments, D2AAK1 can be further developed into a potential treatment for neurodegenerative diseases, in particular related to memory impairment. In summary, D2AAK1 has promising properties for potential treatments of neurodegenerative diseases.

Effectiveness of various methods of DNA isolation from bones and teeth of animals exposed to high temperature.

In the event of fires, natural disasters, and other events associated with high temperature, bones and teeth are the only source of genetic material for identifying human or animal carcasses. To obtain reliable final results of identification tests, the use of appropriate nucleic acid extraction methods is crucial. Therefore, the main objective of this research was to evaluate the effectiveness of selected methods of DNA isolation from animal burnt bones and teeth. In addition, the effect of the duration of high temperature on the stability of nuclear and mitochondrial DNA in these tissues was determined, as well as the possibility of using the genetic material obtained for species identification of remains of unknown origin. Bones and teeth collected during necropsy of dogs were burnt in a laboratory oven at 400 °C (752 °F; 673.15 K) for 5, 10, 15, 30, 45 and 60 min. DNA was isolated according to four different protocols, using three commercial kits, i.e. the PrepFiler® Forensic DNA Extraction Kit from Applied Biosystems, the QIAamp® DNA Investigator Kit from QIAGEN, and the DNA Mini Kit from Syngen, as well as a classic organic method. The effectiveness of these methods was compared by assessing the amount of isolated DNA using Real-Time PCR and its purity using a NanoDrop™ spectrophotometer. Each isolate was also subjected to PCR with primers designed to amplify fragments of dog mitochondrial DNA. The effectiveness of species identification was assessed for the method showing the best DNA recovery and for the organic method, considered the gold standard for analysis of difficult material. The QIAamp® DNA Investigator Kit showed the highest efficiency of DNA isolation from bones and teeth burnt for 15 min (the longest burning time for which DNA could still be recovered from bones and teeth). The results of the experiment clearly indicate that DNA stability in hard tissues depends on how long they burn. In the case of exposure to 400 °C, reliable genetic testing, including species identification, is possible when the burning time does not exceed 15 min. Among the hard tissues examined, bones proved more suitable than teeth for identification purposes. It was also concluded that identification of bone remains with extreme heat damage should be based on mitochondrial DNA analysis.

COVID-19 in the autopsy room-requirements, safety, recommendations and pathological findings.

Modern technologies enable the exchange of information about the expansion of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and the continually increasing number of the coronavirus disease 2019 (COVID-19) cases almost in real time. The gravity of a current epidemiological situation is represented by the mortality rates, which are scrupulously updated daily. Performing autopsies on patients with either suspected or confirmed COVID-19 is of high importance since these might not only improve clinical management but also reduce the risk of SARS-CoV-2 infection expansion. The following paper aimed to present the most crucial aspects of SARS-CoV-2 infection from the point of view of forensic experts and pathologists, recommendations and safety precautions regarding autopsies, autopsy room requirements, possible techniques, examinations used for effective viral detection, recommendations regarding burials, and gross and microscopic pathological findings of the deceased who died due to SARS-CoV-2 infection. Autopsies remain the gold standard for determining the cause of death. Therefore, it would be beneficial to perform autopsies on patients with both suspected and confirmed COVID-19, especially those with coexisting comorbidities.

Post-mortem analysis of gunshot wounds to the head and thorax in dogs by computed tomography, radiography and forensic necropsy.

In view of the scarcity of literature data on the use of radiological imaging techniques in forensic veterinary medicine, while at the same time the number of reported crimes against animals involving the use of firearms is rising, this paper attempts to assess the usefulness of radiography and computed tomography (CT) in the post-mortem diagnosis of gunshot wounds (GSW) in comparison to classic necropsy. The design of the experiment was as follows: preparation of the research material (13 dog carcasses), shooting of the material from different distances (1.5 and 12 m, plus one contact shot to the head) and using different types of ammunition, followed by X-rays and CT scans in each case to examine the injuries resulting from the shot. The final steps of the experiment were photographic documentation and autopsy by the Virchow method. In the examined material, post-traumatic bone lesions and the presence of metallic foreign bodies were successfully imaged by both radiography and CT. GSW analysis using CT provided much better data quality and some additional information. Two general conclusions can be drawn from the results of the experiment. First, damage caused by gunshots is correlated with the calibre, initial velocity and kinetic energy of the projectile, as well as the distance from the muzzle of the gun to the object shot. Second, radiological examination is useful in preparing forensic veterinary opinions. Used as a complement to classic necropsy, they increase the possibility of an accurate post-mortem diagnosis of shooting victims.

The effect of the high-fat diet supplemented with various forms of chromium on rats body composition, liver metabolism and organ histology Cr in liver metabolism and histology of selected organs.

BACKGROUND: In the present study, we hypothesized that feeding rats a high-fat diet negatively affects liver metabolism and function and disturbs the histology of some internal organs. We also postulated that there is a form of chromium whose administration alleviates the negative effects of a high-fat diet in rats. METHODS: To verify the hypotheses, we tested the effect of various forms of chrome (picolinate - Cr-Pic, Chromium(III)-methionine complex - Cr-Met, and chrome nanoparticles - Cr-NPs) applied in the recommended amount of 0.3 mg/kg of BW on growth parameters, body fat, liver metabolism and functional disorders, and histological parameters of selected internal organs in rats fed a standard (S) or high-fat diet (F). The experiment was conducted on 56 male outbred Wistar rats (Rattus norvegicus. Cmdb:WI) randomly divided into eight experimental groups. For eight weeks the rats received a standard or high-fat diet, without Cr or with Cr at 0.3 mg/kg diet in the form of Cr-Pic, Cr-Met or Cr-NPs. RESULTS AND CONCLUSION: The use of a F diet disrupted the lipid-carbohydrate profile, worsened liver metabolism and function, reduced the expression of hepatic PPAR-α and leaded to negative changes in the histological image of internal organs - liver, kidneys and pancreas. The 8-week use of an chromium supplement in a F diet, regardless of the form used, did not improve the ratio of fat tissue to lean tissue, worsened liver function and negatively affected on the histological image of the liver, kidneys and pancreas. However, the most negative changes in lipid-carbohydrate metabolism and liver functioning were observed with CrNPs supplementation.

Antioxidant Status and Liver Function of Young Turkeys Receiving a Diet with Full-Fat Insect Meal from Hermetia illucens.

We hypothesized that full-fat insect meal from Hermetia illucens (HI) larvae can be an acceptable source of protein and energy in the diet of young turkeys, in an amount adapted to the nutritional needs of these birds, and at the same time can improve their antioxidant status and metabolism. The turkeys were fed a control diet (HI0) without the insect meal, and three diets with increasing HI content of 5%, 10% and 15% (treatments HI5, HI10 and HI15, respectively). The use of 10% or 15% HI in the diet of young turkeys, while beneficially raising levels of P, Fe and Hb, has a negative effect on lipid metabolism, increasing TC levels, lipid oxidation, and fat deposition in the liver. The inclusion of 5% HI in the diet of young turkeys has no adverse effect on the lipid status and histology of the liver, but it does not improve antioxidant status. To conclude, the level of HI meal in the diet of turkeys should not exceed 5%. However, as similar studies on turkeys have not yet been published, overly general conclusions should not be drawn from the results of the present study, and further research is necessary.

Assessment of DNA Methylation and Oxidative Changes in the Heart and Brain of Rats Receiving a High-Fat Diet Supplemented with Various Forms of Chromium.

The aim of the study was to determine how feeding rats a high-fat diet supplemented with various forms of chromium affects DNA methylation and oxidation reactions as well as the histology of heart and brain tissue. The rats received standard diet or high-fat diet and chromium at 0.3 mg/kg body weight (BW) in form of chromium (III) picolinate, chromium (III)-methionine, or nano-sized chromium. The content of malondialdehyde (MDA), protein carbonyl (PC), and 8-hydroxydeoxyguanosine (8-OHDG), the level of global DNA methylation and the activity of selected DNA repair enzymes were determined in the blood. In the brain and heart, the content of MDA, PC, 8-OHDG, and levels of global DNA methylation were determined. The brain was subjected to histological examination. The use of a high-fat diet was found to intensify epigenetic changes and oxidation reactions in the heart and brain. It was concluded that epigenetic changes and oxidation of lipids, proteins, and DNA in the heart and brain of rats resulting from the use of a high-fat diet cannot be limited by supplementing the diet with chromium. It was established that the use of chromium to supplement a high-fat diet intensifies the negative epigenetic and oxidative changes in the heart and brain, especially in the case of chromium nanoparticles.

The role of linagliptin, a selective dipeptidyl peptidase-4 inhibitor, in the morphine rewarding effects in rats.

Linagliptin is a selective dipeptidyl peptidase-4 (DPP-4) inhibitor which suppresses the rapid degradation of endogenous glucagon-like peptide-1 (GLP-1). In clinical practice, it is used as an antidiabetic drug, but recent studies have confirmed its role in the activity of the central nervous system (CNS). The reported study focused on the role of linagliptin (10 and 20 mg/kg, ip) in the morphine rewarding effect, analyzing how the agent had influenced the conditioned place preference (CPP) in rats via the expression, acquisition, extinction and reinstatement of the morphine rewarding effect. The obtained results clearly demonstrated linagliptin to inhibit the expression and acquisition, to accelerate the extinction and, eventually, to reduce the reinstatement of morphine-induced CPP. The undertaken experiments significantly extended our knowledge on the mechanisms behind the morphine rewarding effect.

SB-334867 (an Orexin-1 Receptor Antagonist) Effects on Morphine-Induced Sensitization in Mice-a View on Receptor Mechanisms.

The present study focused upon the role of SB-334867, an orexin-1 receptor antagonist, in the acquisition of morphine-induced sensitization to locomotor activity in mice. Behavioral sensitization is an enhanced systemic reaction to the same dose of an addictive substance, which assumingly increases both the desire for the drug and the risk of relapse to addiction. Morphine-induced sensitization in mice was achieved by sporadic doses (five injections every 3 days) of morphine (10 mg/kg, i.p.), while a challenge dose of morphine (10 mg/kg) was injected 7 days later. In order to assess the impact of orexin system blockade on the acquisition of sensitization, SB-334867 was administered before each morphine injection, except the morphine challenge dose. The locomotor activity test was performed on each day of morphine administration. Brain structures (striatum, hippocampus, and prefrontal cortex) were collected after behavioral tests for molecular experiments in which mRNA expression of orexin, dopamine, and adenosine receptors was explored by the qRT-PCR technique. Additionally, the mRNA expression of markers, such as GFAP and Iba-1, was also analyzed by the same technique. SB-334867 inhibited the acquisition of morphine-induced sensitization to locomotor activity of mice. Significant alterations were observed in mRNA expression of orexin, dopamine, and adenosine receptors and in the expression of GFAP and Iba-1, showing a broad range of interactions in the mesolimbic system among orexin, dopamine, adenosine, and glial cells during behavioral sensitization. Summing up, the orexin system may be an effective measure to inhibit morphine-induced behavioral sensitization.

The adenosinergic system is involved in sensitization to morphine withdrawal signs in rats-neurochemical and molecular basis in dopaminergic system.

RATIONALE: Experimental data informs that not only do the dose and time duration of dependent drugs affect the severity of withdrawal episodes. Previous withdrawal experiences may intensify this process, which is referred as sensitization to withdrawal signs. Adenosine and dopamine (DA) receptors may be involved in this sensitization. OBJECTIVES: Rats were continuously and sporadically treated with increasing doses of morphine for 8 days. In rats, sporadically treated with morphine, morphine administration was modified by adding three morphine-free periods. Adenosine agonists were given during each of the morphine-free periods (six injections in total). On the 9th day, morphine was injected. One hour later, naloxone was administered to induce morphine withdrawal signs. Then, the animals were placed into cylinders and the number of jumpings was recorded. Next, the rats were decapitated and brain and brain structures (striatum, hippocampus, and prefrontal cortex) were dissected for neurochemical, molecular, and immunohistochemical experiments within DAergic pathways. RESULTS: We demonstrated that previous experiences of opioid withdrawal intensified subsequent withdrawal signs. Adenosine ligands attenuated the sensitization to withdrawal signs. In a neurochemical study, the release of DA and its metabolites was impaired in all structures. Significant alterations were also observed in mRNA and protein expression of DA receptors. CONCLUSIONS: Results demonstrate that intermittent treatment with morphine induces alterations in the DAergic system which may be responsible for sensitization to morphine withdrawal signs. Although adenosine ligands attenuate this type of sensitization, they are not able to fully restore the physiological brain status.

Effects of perinatal exposure to lead (Pb) on purine receptor expression in the brain and gliosis in rats tolerant to morphine analgesia.

The aim of the present study was to investigate the molecular effects of perinatal exposure to lead (Pb) on protein and mRNA expression of purine receptors: P2X4, P2X7, adenosine receptor A1; and astrocytes (GFAP mRNA expression) and on microglia activation (Iba1 mRNA expression) in several structures of the mesolimbic system (striatum, hippocampus, prefrontal cortex) in rats expressing tolerance to the antinociceptive effect of morphine. Rat mothers were orally treated with 0.1% lead acetate from conception, through gestation, and postnatally, as well as to offspring up to day (PND) 28; subsequently molecular studies were conducted on adult (PND 60) male rats. Morphine tolerance developed more strongly in rats perinatally exposed to Pb. The analysis revealed a significant up-regulation of protein and mRNA P2X4 receptor expression in the striatum and prefrontal cortex but not in the hippocampus; P2X7 protein and mRNA receptor expression in the striatum and hippocampus, but not in the prefrontal cortex; A1 protein receptor expression in all investigated structures and A1 mRNA expression in the striatum and hippocampus; Iba1 mRNA expression in the striatum and hippocampus; and GFAP mRNA expression in the striatum and prefrontal cortex. Immunohistochemical analysis has also revealed significant alterations. Strong expressions of P2X4, P2X7, A1 receptors, astrocytes and microglia activation were observed in the hippocampus in Pb and/or morphine treated rats. The higher expression of purine receptors and glial cell activation are important markers of neuroinflammatory processes. Therefore, we conclude that Pb-induced neuroinflammation may be responsible for the intensification of morphine tolerance in the Pb-treated rats. Additionally, the dysregulation of A1 adenosine receptors, mainly in the hippocampus, may also be involved in the intensification of morphine tolerance in Pb-treated rats. Our study demonstrates the significant participation of environmental factors in addictive process; additionally, it shows the necessity of modification of addictive disorder with neuroprotective agents.

Effects of the adenosinergic system on the expression and acquisition of sensitization to conditioned place preference in morphine-conditioned rats.

In the presented study, we attempt to investigate if the sensitization to conditioned place preference (CPP) induced by low doses of morphine was developed in rats which have been previously conditioned with morphine. The experiments were performed in the CPP test. Firstly, it has been demonstrated that administration of ineffective dose of morphine on the 9th day induces the increase in time spent of rats at a morphine-paired compartment, confirming that sensitization to CPP has been developed in these animals. Secondly, it has been shown that stimulation of A1 receptor significantly inhibits the expression of morphine-induced of sensitization, and blockade of these receptors produces the opposite effect. Finally, it has been indicated that both stimulation and blockade of A1 and/or A2A receptors inhibit the acquisition of sensitization to CPP. The obtained results have strongly supported the significance of adenosinergic system in both expression and acquisition of studied sensitization. These results seem to be important for the identification of connections in the central nervous system which can help finding new strategies to attenuate rewarding action of morphine.

Analysis of cases of forensic veterinary opinions produced in a research and teaching unit.

The aim of the study was to present the results of necropsies carried out in the years 2000-2014 in the Department of Pathological Anatomy, Faculty of Veterinary Medicine, University of Life Sciences in Lublin. The material used for the analysis consisted of expert opinions prepared on the basis of a decision by a judicial body to admit an expert opinion as evidence. An increase was observed in the demand for the services of veterinary forensic experts, beginning in 2006 and persisting through 2014. The response to the growing popularity of veterinary forensic examinations should be systematization of knowledge and exchange of experience, which would enable the further development of this interdisciplinary science.

The effect of perinatal lead exposure on dopamine receptor D2 expression in morphine dependent rats.

The aim of this study was to investigate the behavioral and molecular effects of pre- and postnatal lead (Pb) exposure on the expression of morphine withdrawal and tolerance in adult rats. Rats were orally treated with 0.1% (1000ppm) lead acetate from conception, through gestation, up to postnatal day (PND) 28. Subsequently, behavioral experiments were conducted on adult (PND 60) male rats. To assess behavioral effects of morphine dependence in Pb-exposed rats two experimental models were used: naloxone-precipitated withdrawal signs and the assessment of morphine tolerance to antinociceptive effect in the tail-immersion test. Morphine withdrawal and tolerance were more expressed in Pb-exposed morphine administered rats than in morphine administered rats. In the case of morphine withdrawal signs the analysis of protein (Western blotting) and mRNA (RT PCR) expression revealed significantly higher dopamine D2 receptor (D2R) expression in prefrontal cortex, but not in striatum and hippocampus, in Pb-exposed morphine administered rats than in morphine administered rats. Differently, in the case of morphine tolerance the significant upregulation of D2R protein and mRNA expression in hippocampus, but not in prefrontal cortex or striatum, was demonstrated in Pb-exposed and morphine administered rats in comparison with morphine administered. These findings suggest that in morphine withdrawal and tolerant rats the perinatal Pb-exposure can affect D2R expression in brain region-specific manner. Immunohistochemical assessment of D2R expression in hippocampus showed translocation of D2R from membrane-cytoplasm in control rats to nucleus in morphine administered rats. Perinatal Pb-exposure did not induce the changes in the localization of D2R irrespective of morphine effect.

Granulomatous peritonitis in a European brown bear caused by Baylisascaris transfuga.

We report a case of granulomatous peritonitis due to Baylisascaris transfuga in a young male European brown bear (Ursus arctus). At necropsy, there were extensive abdominal adhesions and extensive granulomatous tissue on the peritoneum and liver capsule. In the gastrointestinal tract, there were 58 nematodes that were identified as Baylisascaris transfuga using light and scanning electron microscopy.