The Influence of Copper Nanoparticles on Neurometabolism Marker Levels in the Brain and Intestine in a Rat Model.

The aim of this study is to assess the effect of different forms and dosages of copper on the levels of markers depicting the neurodegenerative changes in the brain and the jejunum. The experiment was performed using 40 male Wistar rats fed a typical rat diet with two dosages of Cu used as CuCO3 (6.5 and 13 mg/kg diet) and dietary addition of two CuNP dosages (standard 6.5 and enhanced 13 mg/kg diet), randomly divided into four groups. The levels of neurodegenerative markers were evaluated. Nanoparticles caused a reduction in the level of glycosylated acetylcholinesterase (GAChE), an increase the level of acetylcholinesterase (AChE) and lipoprotein receptor-related protein 1 (LRP1), a reduction in ß-amyloid (ßAP) in the brain and in the intestine of rats and a reduction in Tau protein in the brain of rats. The highest levels of AChE, the ATP-binding cassette transporters (ABC) and LRP1 and lower levels of toxic GAChE, ß-amyloid, Tau, hyper-phosphorylated Tau protein (p-Tau) and the complex of calmodulin and Ca2+ (CAMK2a) were recorded in the tissues of rats receiving a standard dose of Cu. The neuroprotective effect of Cu can be increased by replacing the carbonate form with nanoparticles and there is no need to increase the dose of copper.

Dietary Copper Deficiency Leads to Changes in Gene Expression Indicating an Increased Demand for NADH in the Prefrontal Cortex of the Rat's Brain.

Copper is an essential element to brain cells as it is a cofactor and a structural component of various enzymes involved in energy metabolism pathways. Accumulating evidence points to the pivotal role of copper deficiency in neurodegeneration resulting from impaired copper homeostasis. Despite the indisputable role of copper in mitochondrial respiration, its homeostasis regulation in the brain tissue remains unclear. The assessment of changes in the expression of genes encoding key pathways of energy metabolism can greatly benefit further studies exploring copper's role in neurodegeneration. Using a rat model, we investigate whether the replacement of the inorganic form of copper with metallic nanoparticles containing copper or complete deprivation of copper from the diet have an impact on the expression of genes involved in energy metabolism in the prefrontal cortex of the rats' brain. Herein, we indicate that removing inorganic copper from the normal standard diet or the replacement with copper nanoparticles can lead to programmed energy metabolism changes. It can be recognized that some of these changes indicate an increased demand for NADH in the prefrontal cortex of the rat's brain, probably as a result of adaptation effect.

Selected morphological and mechanical features of the femur of pregnant rats after exposure to caffeine applied at various temperatures Caffeine in the nutrition of pregnant rats.

It was assumed that the temperature at which caffeine is consumed may cause changes in the mechanical and morphological properties of bones and affect the bone metabolism of pregnant female. The aim of the study was to assess the effect of caffeine used at different temperatures on selected morphological and mechanical properties of the femur as well as biochemical indicators of bone formation in female rats in pregnancy. It was use test solution at the temperature 10, 25 and 45oC from 8 to 21 day during pregnancy, once a day. It was found that the temperature of administrated caffeine may have an effect on changing the morphometric properties and on the bone metabolism of pregnant female rats. The application of caffeine solution, was administrated at 10oC caused the most constrained growth of the femur and weakened resistance to load, and caused increased susceptibility to cracks. Analysis of bone metabolism indicators showed that caffeine administered in the form of a solution at 10oC and 25oC caused the most negativity effects for bone formation and bone turnover indicators. The administration of caffeine at 10oC causes the largest negative changes in bone morphological and strength indicators and hasn't a beneficial effect on the bone metabolism of pregnant female rats.

The effect of the use of copper carbonate and copper nanoparticles in the diet of rats on the level of ß-amyloid and acetylcholinesterase in selected organs.

BACKGROUND: Copper has an important role in nervous system function, as a cofactor of many enzymes and in the synthesis of neurotransmitters. Both the dose and the chemical form of copper can determine the impact of this element on metabolism, the neurological system and the immune system. AIMS: The aim of the study was to determine whether and in what form the addition of copper changes the level of amyloid beta and acetylcholinesterase level in selected rat tissues. METHODS: Thirty, healthy, male, albino Wistar rats aged 7 weeks were randomly divided into 3 groups. Three experimental treatments were used to evaluate the effects of different levels and sources of Cu (6.5 mg kg of diet) in the diet: Cu0 - rats fed a diet without Cu supplementation; Cusalt - rats fed a diet with CuCO3 (6.5 mg kg of diet) during two months of feeding; CuNPs - rats fed a diet with Cu nanoparticles (6.5 mg kg of diet) during two months of feeding. In blood serum and tissue homogenates there rated the indicators proving the potential neurodegenerative effect and epigenetic DNA damage induced by chemical form of copper or lack of additional copper supplementation in diet were determined. There were analysed: level of acetylcholinesterase, ß-amyloid, low-density lipoprotein receptor-related protein 1, apyrimidinic endonuclease, thymidine glycosidase, alkylpurine-DNA-N-glycosylase and glycosylated acetylcholinesterase. RESULTS: Irrespective of the form of copper added, it was found to increase acetylcholinesterase level in the brain, spleen and liver, as well as in the blood plasma of the rats. Copper in the form of CuCO3 was found to increase acetylcholinesterase level in the kidneys. The addition of both forms of copper caused a marked increase in the plasma concentration of ß-amyloid in comparison with the diet with no added Cu. The addition of both forms of copper caused a marked increase in the plasma concentration of ß-amyloid in comparison with the diet with no added Cu. CONCLUSIONS: A lack of added Cu in the diet of rats reduces the concentration of amyloid-ß in the blood, whereas administration of copper, in the form of either CuNPs or CuCO3, increases the level of this peptide in the blood. The use of copper in the form of CuNPs in the diet of rats does not increase the level of ß-amyloid more than the use of the carbonate form of this element. The use of CuNPs or CuCO3 in the diet of rats increases acetylcholinesterase level in the brain, spleen, liver, and blood. CuNPs in the diet of rats were not found to increase acetylcholinesterase level to a greater extent than Cu+2 carbonate.

The Effect of Administration of a Phytobiotic Containing Cinnamon Oil and Citric Acid on the Metabolism, Immunity, and Growth Performance of Broiler Chickens.

It was postulated that a phytobiotic preparation containing cinnamon oil and citric acid added to drinking water for chickens in a suitable amount and for a suitable time would beneficially modify the microbiota composition and morphology of the small intestine, thereby improving immunity and growth performance without inducing metabolic disorders. The aim of the study was to establish the dosage and time of administration of such a phytobiotic that would have the most beneficial effect on the intestinal histology and microbiota, production results, and immune and metabolic status of broiler chickens. The experiment was carried out on 980 one-day-old male chickens until the age of 42 days. The chickens were assigned to seven experimental groups of 140 birds each (seven replications of 20 individuals each). The control group (G-C) did not receive the phytobiotic. Groups CT-0.05, CT-0.1, and CT-0.25 received the phytobiotic in their drinking water in the amount of 0.05, 0.1, and 0.2 mL/L, respectively, at days 1-42 of life (continuous application, CT). The birds in groups PT-0.05, PT-0.5, and PT-0.25 received the phytobiotic in the same amounts, but only at days 1-7, 15-21, and 29-35 of life (periodic application, PT). Selected antioxidant and biochemical parameters were determined in the blood of the chickens, as well as parameters of immune status and redox status. The morphology of the intestinal epithelium, composition of the microbiome, and production parameters of chickens receiving the phytobiotic in their drinking water were determined as well. The addition of a phytobiotic containing cinnamon oil and citric acid to the drinking water of broiler chickens at a suitable dosage and for a suitable time can beneficially modify the microbiome composition and morphometry of the small intestine (total number of fungi p < 0.001, total number of aerobic bacteria p < 0.001; and total number of coliform bacteria p < 0.001 was decreased) improving the immunity and growth performance of the chickens (there occurred a villi lengthening p = 0.002 and crypts deepening p = 0.003). Among the three tested dosages (0.05, 0.1, and 0.25 mL/L of water) of the preparation containing cinnamon oil, the dosage of 0.25 mL/L of water administered for 42 days proved to be most beneficial. Chickens receiving the phytobiotic in the amount of 0.25 mL/L had better growth performance, which was linked to the beneficial effect of the preparation on the microbiome of the small intestine, metabolism (the HDL level p = 0.017 was increased; and a decreased level of total cholesterol (TC) p = 0.018 and nonesterified fatty acids (NEFA) p = 0.007, LDL p = 0.041, as well as triacylglycerols (TAG) p = 0.014), and immune (the level of lysozyme p = 0.041 was increased, as well as the percentage of phagocytic cells p = 0.034, phagocytosis index p = 0.038, and Ig-A level p = 0.031) and antioxidant system (the level of LOOH p < 0.001, MDA p = 0.002, and the activity of Catalase (CAT) p < 0.001 were decreased, but the level of ferric reducing ability of plasma (FRAP) p = 0.029, glutathione p = 0.045 and vitamin C p = 0.021 were increased).

The effect of the source and dosage of dietary Cu on redox status in rat tissues.

The aim of this experiment was to investigate whether the amount of Cu added to the diet of rats can be reduced without adversely affecting the antioxidant status of tissues and growth, and whether copper nanoparticles can be used for this purpose. For four weeks, four experimental groups of rats were fed diets with two dosages of added Cu (standard-6.5 or 3.25 mg/kg) in two forms (standard-CuCO3 or copper nanoparticles). Replacing the CuCO3 supplement with CuNPs resulted in a decreased lung weight and an increased Cu content in brain, kidney and lung, intensification of lipid peroxidation processes, and weakened antioxidant defence in the lungs and kidneys. This treatment also reduced the Cu content in heart, level of lipid oxidation in the liver and testes and improved antioxidant defence in the brain. Reducing the addition of Cu to the diet from 6.5 to 3.25 mg/kg reduced lung weight and increased lipid peroxidation in the liver, heart and lungs, and also weakened antioxidant defence in the lungs and testes. This treatment also weakened the lipid peroxidation process in the spleen, small intestine and brain and strengthened the antioxidant defence of the brain and kidneys. In conclusion, replacing CuCO3 with CuNPs and reducing the level of Cu in the diet of rats has a particularly unfavourable effect on the respiratory system, causing adverse changes in the lungs. However, these treatments have a clearly positive effect on the redox status of the liver and brain.

[Changes of carbohydrate and lipid metabolism in women during pregnancy taking caffeine]. / Zmiany gospodarki weglowodanowej i lipidowej u kobiet w okresie ciazy przyjmujacych kofeine.

Adaptive physiological changes that occur in pregnant woman's body can fluctuate with the intake of substances with proven adverse biological effect on the body. Due to the fact that caffeine is one of the most chronically used xenobiotics it required a research for the impact of consuming caffeine on adaptive processes in the glucose and fatty acid homeostasis of a pregnant woman Many researchers emphasize its negative effect on the glucose and fatty acid homeostasis of the mother and her offspring. However, in spite of years of observation, there is no clear answer to what amount or in what period of time the caffeine modulates the adaptive processes during pregnancy. Because of the potential risk the supply of caffeine during pregnancy should be subjected to considerable restrictions.

[Caffeine and adaptive changes in the circulatory system during pregnancy]. / Kofeina a zmiany adaptacyjne ukladu krazenia u kobiet w okresie ciazy.

Adaptive physiological changes that occur in pregnant women can fluctuate with the intake of substances with proven, adverse biological effect on the body. Due to the fact that caffeine is one of the most chronically used xenobiotics, the impact of consuming caffeine on adaptive processes in the circulatory system of a pregnant women required a research. Many researchers emphasise its negative effect on the circulatory system of the mother and her offspring. However, in spite of years of observation, there is no clear answer to what extent dose or in what period of time the caffeine modulates the adaptive processes during pregnancy. Because of the potential risk the supply of caffeine during pregnancy should be subjected to considerable restrictions.

[Morphological and molecular bases of cardiac development]. / Morfologiczne i molekularne podstawyrozwoju serca.

The heart is a mesoderm-derived organ, whose formation is regulated by various genes. Initially, the most important is expression of Nkx2.5, CR1, pitx2, anf and mhc2a, which are responsible for differentiation of cardiomyocytes. In a later phase activation of mhc2b, pitx2c, mesp1, pcmf1, vmhc, xin, mcl2v, mlc2a, mlc2a, mef2, hand1 and hand2 was revealed. Their expression is regulated by various molecules, including transcription (XIN, GATA, MEF, Tbx5, Baf60c, PECAM, tie-2, MEF2) and growth (VEGF, FGF, PDGF) factors, as well as proteins (i.e., dickkopf-1, cerberus, cytotactin, fibrillin, nodal, thrombomodulin, Wnt, bone morphometric ones - BMP2, BMP 4, BMP5, BMP7) and other substances, such as retinoid and folic acid. Crucial steps in cardiac organogenesis are development of the ventricle and atrial formation, as well as septation and valve formation. Any disturbances of such processes may lead to various congenital heart diseases and defects that could be initiated by various genetic, epigenetic or environmental factors. The most common heart malformations are: stenosis (coarctation) of the aorta and pulmonary trunk, bicuspid aortic valve, atrial and/or ventricular septal defect, persistent truncus arteriosus (Botallo duct), transposition of the great vessels, tricuspid atresia, hypoplastic left and right heart, as well as syndrome of Lutembachera, Cantrell, Ebstein, Eisenmenger and Shone and trilogy, tetralogy, pentalogy of Fallot.

Intensification of doxorubicin-related oxidative stress in the heart by hypothyroidism is not related to the expression of cytochrome P450 NADPH-reductase and inducible nitric oxide synthase, as well as activity of xanthine oxidase.

Cytochrome P450 NADPH-reductase (P450R), inducible synthase (iNOS) and xanthine oxidase play an important role in the antracycline-related cardiotoxicity. The expression of P450R and iNOS is regulated by triiodothyronine. The aim of this study was to evaluate the effect of methimazole-induced hypothyreosis on oxidative stress secondary to doxorubicin administration. 48 hours after methimazole giving cessation, rats were exposed to doxorubicin (2.0, 5.0 and 15 mg/kg). Blood and heart were collected 4, 48 and 96 h after the drug administration. Animals exposed exclusively to doxorubicin or untreated ones were also assessed. The hypothyreosis (0.025% of methimazole) significantly increased the doxorubicin effect on the cardiac carbonyl group and they may increase the glutathione level. An insignificant effect of methimazole was noticed in case of the cardiac lipid peroxidation product, the amount of DNA oxidative damages, iNOS and xanthine oxidase-enzymes responsible for red-ox activation of doxorubicin. However, the concentration of P450R was affected by a lower dose of methimazole in rats administered with doxorubicin. Since in rats receiving doxorubicin changes in oxidative stress caused by methimazole were not accompanied by elevation of bioreductive enzymes, it may be concluded that these changes in the oxidative stress were not related to the tested enzymes.

[Developmental toxicity of the over-the-counter analgetics and antypiretics]. / Toksycznosc rozwojowa leków przeciwbólowych i przeciwgoraczkowych dostepnych bez recepty.

Over-the-counter antipyretics (acetaminophen, aspirin, ibuprofen) and/or analgesics (acetaminophen, aspirin, diclofenac, ibuprofen, naproxen) are relatively safe for adults. However, data on their developmental toxicity is sparse. Moreover, experimental and clinical findings are commonly incompatible. The highest developmental toxicity was found for aspirin. However, unlike full-dose of aspirin, its low doses are compatible during pregnancy. Over-the-counter nonselective cyclooxygenase inhibitors (non-steroidal anti-inflammatory drugs, NSAID) may induce miscarriage, stillbirth, intrauterine growth retardation, preterm constriction of the ductus arteriosus with a secondary persistent pulmonary hypertension, reduced fetal renal perfusion that led to oligohydramion, prolonged pregnancy as well as an increase prevalence of intracranial bleeding in newborns. For acetaminophen and ibuprofen an increased risk of cryptorchidism was also pointed. Prenatal exposure to acetaminophen may also induce bronchial asthma during childhood, preeclampsia, preterm birth, maternal phlebothrombosis and pulmonary embolism. Furthermore, epidemiological data suggest higher risk of cardiac, abdominal wall and orofacial defects of evaluated drugs.

Pathogenicity of the family Legionellaceae.

The Legionellae are Gram-negative bacteria able to survive and replicate in a wide range of protozoan hosts in natural environments, but they also occur in man-made aquatic systems, which are the major source of infection. After transmission to humans via aerosols, Legionella spp. can cause pneumonia (Legionnaires' disease) or influenza-like respiratory infections (Pontiac fever). In children, Legionnaires' disease is uncommon and is mainly diagnosed in children with immunosuppression. The clinical picture of Legionella pneumonia does not allow differentiation from pneumonia caused by others pathogens. The key to diagnosis is performing appropriate microbiological testing. The clinical presentation and the natural course of Legionnaires' disease in children are not clear due to an insufficient number of samples, but morbidity and mortality caused by this infection are extremely high. The mortality rate for legionellosis depends on the promptness of an appropriate antibiotic therapy. Fluoroquinolones are the most efficacious drugs against Legionella. A combination of these drugs with macrolides seems to be promising in the treatment of immunosuppressed patients and individuals with severe legionellosis. Although all Legionella species are considered potentially pathogenic for humans, Legionella pneumophila is the etiological agent responsible for most reported cases of community-acquired and nosocomial legionellosis.

[Occurrence and pathogenicity of the family of Legionellaceae]. / Wystepowanie i chorobotwórczosc bakterii z rodziny Legionellaceae.

Legionella are widespread in natural aquatic environments and are able to exist in water of different temperatures, pH level, and nutrient and oxygen content. Their occurrence in nature can be attributed to their relationships with other microorganisms. When Legionella co-exist with algae and other bacteria, especially in biofilms, the availability of nutrients increases. They also are able to infect protozoa and subsequently reproduce within these organisms. As a result of these relationships, Legionella are protected against adverse conditions, including standard water disinfection techniques. Consequently, they are also prevalent in anthropogenic water such as drinking water, whirlpools, and cooling tower reservoirs. Aerosol-generating systems aid in the transmission of Legionella from water to the air. Human inhalation of contaminated aerosols leads to Legionella infection and disease outbreaks. Legionellosis in humans has typically been characterized as either a potentially fatal pneumonic condition, known as Legionnaires' disease, or an acute, self-limiting, non-pneumonic condition known as Pontiac fever. In addition, Legionella spp. cause extrapulmonary infection, especially in immunosuppressed patients. Timely treatment of Legionnaires' disease is extremely important for the patient's recovery. Although erythromycin has historically been used to treat patients with Legionnaires' disease, newer macrolides and fluoroquinolones are gaining acceptance as the first choice for treatment. Although 52 species of Legionella are known, Legionella pneumophila serogroup 1 is responsible for more than 80% of hospital- and community-acquired cases of Legionnaires' disease.

Gastrointestinal and hepatic toxicity of selective and non-selective cyclooxygenase-2 inhibitors in pregnant and non-pregnant rats.

The aim of the study was to evaluate the toxicity of non-selective (tolmetin, ibuprofen and piroxicam) and selective (DFU) cyclooxygenase-2 inhibitors on pregnant and non-pregnant rats. The drugs were administered orally once (DFU, piroxicam) or three times (tolmetin, ibuprofen) a day from days 8 through 21 of gestation experiment in three doses. The initial dose was similar to the human antiinflammatory one and set as 8.5 mg/kg (tolmetin, ibuprofen), 0.3 mg/kg (piroxicam) and 0.2 mg/kg (DFU). The middle dose was increased 10 times and the highest one 100 times the initial dose. The highest dose for ibuprofen was set at 200mg/kg due to high mortality. On gestation/experimental day 21 animals were sacrificed, blood was collected and abdominal organs were taken for pathological examination. Activity of alanine and asparate aminotransferases and levels of total protein and urea were determined. Stomach, small and large intestines, and liver were grossly and histologically examined. Dose-dependent mortality, signs of gastrointestinal toxicity, and significant changes of biochemical parameters were found in groups exposed to non-selective COX inhibitors in both pregnant and non-pregnant rats. Mild regressive structural hepatic changes were observed. Significant decrease of protein level in non-pregnant rats treated with high DFU dose, and occasionally observed gastrointestinal changes were the only changes noted in groups exposed to the selective COX-2 inhibitor. Tolerability of non-selective COX inhibitors was lower in both pregnant and non-pregnant groups when compared with DFU. Insignificant mortality and histological changes were noted between pregnant and non-pregnant groups.

The variability of the small cardiac vein in the adult human heart.

Direct analyses were performed on 50 hearts submitted for post mortem examination. The remaining hearts were examined by the corrosion method. The aim of the work was to assess the morphology of the small cardiac vein. The small cardiac vein, which runs along the back surface of the coronary sulcus, would usually drain into the coronary sinus at its right side (86% of cases). Cases of the small cardiac vein draining into the middle cardiac vein were also noticed (12%) or directly into the right atrium (1%). In 1% of the preparations it ran along the right margin in the direction of the apex of the heart. In 30% of the corrosion preparations the small cardiac vein was not accessible with the help of corrosion. In 24% of the dissection preparations it was not possible to reach the small cardiac vein with the help of dissection. A statistically significant relationship was observed between the frequency of the presence or absence of the small cardiac vein and the sex of the donor (p > 0.001). In the group examined the percentage of men who did not have the small cardiac vein was 6 times higher than among the woman. There was no evidence for any statistically significant dependence between the frequency of occurrence or of absence of the small cardiac vein and the technique employed.

The influence of the living conditions on the diagnostics process and treatment of bronchial asthma of developmental age based on the author's own material.

Bronchial asthma is one of the most frequent chronic diseases of developmental age. The occurrence of bronchial asthma with children is connected with individual predisposition to development of atopic disease as well as with the presence of specific environmental conditions. Specific factors include food, inhalatory and contact allergens. Non-specific factors include infections, air pollution, and passive smoking. Environment conditions have the significant influence on the course of the disease. The aim of this work is to estimate the frequency of occurring bronchial asthma with children and the influence of living conditions on the diagnosis and treatment processes. The researches were completed on the group of 43 children aged from 8 months to 14 years. The children attended the Paediatric Counselling and complained about chronic cough or symptoms of wheezing breath. The initial diagnosis included taking a history of a patient and his/her family, physical examinations, and supplementary investigation. The final diagnosis was confirmed after a specialist consultation. After introduction of medical treatment, the follow-up examinations were recommended every 4-6 weeks. In the examined group the children with proper birth weight (89%) who were breast-fed at least for 6 months (78%) predominated. In 23% of cases the living conditions were qualified as inconvenient. 63% of children were in a risk of passive smoking in their place of living. In all age groups the children from the families burdened with atopy dominated (98%). In 44% of cases a pet was present in the place of living. In the examined material the highest percentage of children with diagnosis was observed in groups of children aged 7-10. In this group the boys living in difficult conditions prevailed. A high percentage of young children aged 2-3 years were observed. In this group the girls living in good living conditions prevailed. On the basis of the present experience it has been possible to claim that parents of children aged 4-6 have had the highest consciousness of necessity of urgent diagnosis and medical treatment. Treatment of bronchial asthma was introduced at 98% of cases. In the examined group no essential statistical effectiveness of treatment regarding to sex or the living conditions was found. The presented analysis confirms the role of environmental epidemiology in the setting of true relationship between the diagnosis of asthma and the exposure to specific environmental factors.

Variation in morphology of the valve of the great cardiac vein in the human hearts.

The research was carried on 70 hearts of adult humans. The hearts were subjected to direct analyses on the autopsy material. The presence or absence of the valve of the great cardiac vein was analysed with reference to sex. The terminal valve of the great cardiac vein was found in 92% of the examined hearts. To define the location of the valve of the great cardiac vein the osmium of the left atrium oblique vein was treated as the reference point. In 3% of cases the presence of the valve was observed independently of the opening of the left atrium oblique vein. The valve of the great cardiac vein was placed proximally from the opening of the left atrium oblique vein. The valve of the great cardiac vein placed distally in relation to the beginning of the coronary sinus was never observed. The three types of the great cardiac vein valves were found: 1. single, semilunar cusp; 2. endothelial fold; 3. double pouches.