Hexavalent chromium (Cr(VI)) is an environmental pollutant with vast mutagenic and carcinogenic potential. Various past and recent studies confirm the deleterious effects of Cr(VI) in different models, from invertebrates to mammalians. However, there is a lack of studies that comprehensively assess and correlate Cr(VI) accumulation patterns and the resulting physiological responses. Here we used an attractive toxicological model, male Japanese quail (Coturnix japonica), as an alternative probing system to evaluate Cr(VI) accumulation in the vital organs, including the brain, heart, kidneys, liver, and testes after 20 days of exposure to 1.2 μg/mL and 2.4 μg/mL potassium dichromate-K2Cr2O7 ingested in the form of drinking water. The observed effects were correlated with the shift in immune system readiness, hematological indices, serum biochemistry and enzyme activity. Regardless of the exposure dose, the Cr(VI) distribution and accumulation pattern in terms of relative Cr(VI) concentration in tissues was: testes > kidneys > liver > heart > brain. Moreover, Cr(VI) triggered the development of microcytic and hypochromic anemia and reduced the immune system's readiness to cope with challenges. Besides, serum biochemistry presented significant shifts, including reduction of serum electrolytes and proteins and an increase in creatine kinase (CK) and lactate dehydrogenase (LDH) activity. Our study provides novel toxicological data that can be translated to higher animal models to help in the extrapolation of Cr(VI) toxicity in humans.

Tramadol hydrochloride/paracetamol is an opioid that is composed of two different analgesics, tramadol (opioid, 37.5 mg) and paracetamol (non opioid, 325 mg). The study presents first data of tramadol hydrochloride/paracetamol effects on haematological parameters and glucose levels in Wistar rats. Oral administration ad libitum of tramadol hydrochloride/paracetamol was administered during a twenty days period. This research includes two groups of animals. Group I include animals that were administered tramadol hydrochloride/paracetamol daily at dosage of 1.12 g/300 ml and group II that were administered daily at dosage of 1.68 g/300 ml of water. We analayzed 14 haematological parameters and glucose concentration. Significant changes were established for all analyzed parameters. Significantly low numbers of erythrocytes, leukocytes and lymphocytes were observed. Tramadol hydrochloride/paracetamol has an extremely negative effect on haematological parameters in Wistar rats, particularily on the blood coagulation due to the thrombocitopenia, anaemia and weakened immunity. If properly administered, tramadol hydrochloride/paracetamol can be an effective analgesic but at high dosage and over a prolonged period it may cause adverse effects in Wistar rats.

BACKGROUND Lead and cadmium are significant environmental pollutants that cause pathophysiological responses in many organs. Heavy metal absorption into many tissues is very fast due to a pronounced affinity for metallothioneins. METHOD Japanese quail were exposed to different concentrations of metals (cadmium 0.20 mg/L and lead 0.25 and 0.50 mg/L) for 20 days. Erythrocytes (normal and hemolyzed) and lymphocytes (normal and altered) were monitored in this study. The analysis observed the percentage of normal and altered cells, as well as erythrocyte surface area. Cell counts were analyzed using light microscopy, while surface area and cytological changes in cells and nuclei were analyzed using licensed software. RESULTS Different concentrations of metals have caused erythrocyte hemolysis as well as structural and morphological alterations in lymphocytes. Destruction of cell and nucleus membrane, changes in cell size, erythrocyte denucleation and reduced erythrocyte surface area were observed. Cadmium has caused erythrocyte hemolysis (29.30 %) and lymphocyte damage (92.10 %). Higher doses of lead resulted in greater damage to lymphocytes (63 %). Also, treatment with higher dose of lead produced a higher percentage of hemolyzed erythrocytes (19.20 %) in comparison to lower dose (9.90 %). CONCLUSION The toxicity of heavy metals leads to reduced maturation of the blast, which causes the appearance of immature cells in peripheral circulation and severe destruction of blood cell membranes. Erythrocyte hemolysis can lead to anemia, while lymphocyte damage can lead to lymphocytopenia.

Abstract Methadone eliminates heroin use, reduces death rates and criminality associated with heroin use, and improves patients’ health and social productivity. This study included long-term addicts who completed a methadone therapy program as well as relapsed patients. Liver and renal markers important for methadone metabolism were analyzed. Renal markers included urea and creatinine, while hepatic markers included total bilirubin, AST, ALT, γGT, and LDH as nonspecific but significant parameters of liver metabolism. The study included 34 male and 6 female heroin-dependent patients undergoing a rehabilitation program with methadone maintenance treatment (MMT). During therapy, average values of all parameters remained within the reference interval but individual parameters in some patients were very high. Significant differences for urea (0.00) and very high individual variations in all parameters, especially γGT and LDH, were found in patients who were in relapse. Age of the patients did not show a correlation with the presence of significant differences in serum biochemical parameters during therapy. Prolonged use of methadone therapy stabilizes high variations of liver and renal markers. MMT achieves a stabilization of serum indicators relevant for methadone metabolism that correlates with the duration of consumption and the type of opioid substance. The most important hepato-renal markers as indicators of therapy success are γGT, LDH, and creatinine. The validity of former enzymatic tests (AST, ALP, and ALT) should be seriously reconsidered in terms of MTT treatment success and monitoring the health of heroin addicts.

Abstract Cadmium is a heavy metal, toxic even in trace amounts and its biological function in the human body has not been described to date. It is assumed that cadmium manifests in dose-dependent genotoxic and cytotoxic effects on many organs and tissue types. In this study, we have analyzed the biochemical parameters in the serum of Japanese quails (Coturnix japonica) after chronic in vivo exposure to cadmium. Adult animals were exposed to cadmium in the form of CdCl2 dissolved in water (0.20 mg/L) for 20 days. Significant differences between controls and exposed animals were found in 12 out of 13 analyzed biochemical parameters. Total bilirubin concentrations did not show any significant differences between the two groups. Exposure to cadmium has resulted in a significant increase in lactate dehydrogenase activity, sodium and chloride concentration, as well as significant reductions in total proteins, albumins, globulins, glucose, triglycerides, cholesterol, calcium concentration, and alkaline phosphatase activity. In this sense, chronic in vivo exposure to low doses of cadmium has induced severe changes in the levels of observed biochemical parameters and enzyme activity. Additionally, evident cytogenetic changes in the liver were also noted, where hepatocyte damage and even lack of organized nuclei, including nuclear fragmentation, clearly indicated ongoing apoptotic processes.

Cadmium is a heavy metal classified as an environmental hazard, and its toxicity is subject to extensive research. Japanese quails were exposed to cadmium chloride (CdCl2) ad libitum for 20 days. Bone marrow, peripheral blood and liver were analyzed following the exposure. Moreover, we have provided the very first explanation of hematopoietic lines in Japanese quail. Following CdCl2 exposure, changes in the number, size and morphology of blood cells were observed in both peripheral blood and bone marrow. Alterations included severe erythrocyte damage, monocytosis and lymphopenia. In the liver of Cd-exposed animals we observed necrotic cells, absence of hematopoietic regions and cytogenetic changes of hepatocytes. Alterations in the bone marrow were also noted, as well as giant phagocytic cells, most likely macrophages. In vivo, CdCl2 exposure caused swift and destructive changes in the hematopoietic niche, liver and other tissues responsible for the detoxification cycle of cadmium and its compounds.

Copper sulfate has been used for many years as an algaecide and parasite treatment. The usage of copper has certain issues as there is a thin line that separates effective treatment levels from overdoses. Copper sulfate can be extremely toxic to fish under certain conditions. This study is focused on the effects of copper sulfate on some blood indices in Oncorhynchus mykiss during a 24 hour period. Our research used one procedure 40 Oncorhynchus mykiss with the average weight of 220 ±10 g. The experimental group had 20 fish that were treated with the blue vitriol (dose of 0,012 g/40l), during a 24 hour period. The control group had 20 fish. The presence of copper in the water leads to the significant increase of erythrocyte, hemoglobin, MCHC and leukocyte levels while the MCV levels were noted to be considerably low. Monocytes, unsegmented and segmented granylocites were significantly increased in the experimental group of fish. Lymphocyte count was considerably reduced in the same group of fish.

Objectives This study aims to investigate the low-resolution human leukocyte antigen (HLA)-B locus polymorphisms between unrelated healthy individuals and patients with diagnosis of seronegative spondyloarthropathies and determine risky and protective allelic groups and genotypes. Patients and methods The study included 104 healthy control individuals (52 males, 52 females; median age 43 years; range 2 to 76 years) and 96 patients (43 males, 53 females; median age 28.5 years; range 2 to 67 years) diagnosed with: ankylosing spondylitis (AS) (n=19), reactive arthritis (n=19), psoriatic arthritis (n=28) and undifferentiated spondyloarthropathies (n=30). Genomic deoxyribonucleic acid was extracted from peripheral blood to detect allelic groups of HLA class I and II. Single-specific-primer polymerase chain reaction was used for HLA genotyping and visualization of products after their separation on 1.5% agarose gel for horizontal gel electrophoresis. Results Significantly increased frequency was found for HLA-A*02 and HLA-B*27 allelic variants in all groups of patients. The increased frequency of the HLA-B*35 allelic group in the control group represents the protective gene variant for the occurrence of AS. The predisposing genotype (HLA-B*27/B*44 and B*27/B*51) for the onset of disease was only found in AS patients. Conclusion This study shows the strong association of HLA-B*27 antigen with spondyloarthropathies, which is considered a risk variant of the gene for the onset of disease. Protective and risky allelic variants and genotypes are rare and their detection as well as increased frequency are possible if larger numbers of patients are involved.

The main circulatory medium of echinoderms is the coelomic fluid. Biochemistry of coelomic fluid is very complex and slightly different from seawater. The main aim of this research was to analyse concentration of electrolytes (potassium, calcium, magnesium, chlorine and sodium), heavy metals (lead, chromium, cadmium and cobalt) and iron in coelomic fluid of sea urchin, Arbacia lixula as an indirect indicator of seawater contamination. After precise statistical evaluation it was observed that electrolyte concentrations were; Na 189.20±11.41 mmol/l, Cl 134.06±37.08 mmol/l, Mg 4.24±1.08 mmol/l, Ca 3.04±0.84 mmol/l. Biochemical content of heavy metals in coelomic fluid was; Co 1.292±0.879 ppm, Pb 0.644±0.131 ppm, Cr 0.116±0.055 ppm, Cd 0.031±0.017 ppm and iron 0.259±0.058 ppm, and it depends on the potential accumulation level in the organism. The composition of electrolyte and heavy metal content varies depending on the composition of seawater. Obtained values are within the range of values most commonly found in low polluted areas of the Adriatic sea. Similar models may be applied for monitoring of heavy metal contamination in marine environment.

Thermal changes in water cause many metabolic changes that manifest themselves in physiological fish adaptations. The analysis of hematologic and biochemical blood parameters provides important information on environmental influences on the health status of fish. The hematocrit (HCT) (l/l), hemoglobin concentration (Hb) (g/l), mean corpuscular volume (MCV) (fl), mean corpuscular hemoglobin (MCH) (pg), mean corpuscular hemoglobin concentration (MCHC) (g/l) and red blood cells (RBC) (x1012/L) were analyzed. Animals were grouped into two groups: control (n=10) and experimental (n=16). The experimental fish were exposed to 28°C for 30 min. Puncture of the heart was done and the blood without anticoagulant was analyzed. During hyperthermia, an increase in hematological parameters was observed, except for MCV values that were low. Significant differences were established only for the number of erythrocytes and the hematocrit values (p<0.05). The results showed a decrease in MCV and an increase in the value of other erythrocyte parameters. Significant changes in the number of erythrocyte and hematocrit values were found. Some hematological parameters such as erythrocyte and MCV values are significant stress indicators and can serve us as important factors for physiological adaptations of fish. The carp shows excellent ability to adjust to temperature variations that can be seen through the analysis of hematological status.

Abstract Emperipolesis is considered a physiological phenomena often present in various pathophysiological conditions, but its etiology is still unknown. In this study, we analyzed the number of megakaryocytes and the percentage of emperipoletic cells in the sternal and femoral bone marrow of Wistar rats. Five types in the thrombopoiesis lineage (megakaryoblasts, promegakaryocytes and megakaryocytes - acidophilic, basophilic and thrombocytogenic) were determined. Except for basophilic megakaryocytes, significant differences were found for number of thrombopoietic cells in the sternal and femoral bone marrow. A larger number of thrombocytogenic megakaryocytes were present in the sternal bone marrow. Emperipoletic cells were significantly present in the femoral compared to the sternal bone marrow. Emperipolesis was typical for lymphocytes and neutrophils individually, while emperipolesis with two or more cells within thrombopoietic cell was also present (1-7 %) and significant differences between the sternal and femoral bone marrow were detected. Emperipolesis was found in all analysed rats and it most commonly occured within mature megakaryocytes and rarely megakaryoblasts, while it was not recorded in the promegakaryocytes. The high incidence of megakaryocytes with emperopolesis in rats could be a consequence of “normal” cell retention in the cytoplasm of megakaryocytes while passing blood cells to circulation or related to haematopoietic response due to high incidence of inbreeding.