Effect of the treatment stage on the serum levels of selected cytokines and antioxidant enzymes in patients with tumors of the reproductive organs.

OBJECTIVE: Chronic inflammation along with concomitant oxidative stress contributes to an increased risk of cancer development. The aim of this study was to analyze selected cytokines and antioxidant enzymes in patients with ovarian and endometrial cancers, taking into account the stage of oncological treatment. PATIENTS AND METHODS: The study sample included 52 female patients with advanced endometrial cancer (n = 26.50%) and ovarian cancer (n = 26.50%), undergoing chemotherapy. Long-term observation at four time points was used in the subjects. Each of the women was blood sampled several times (before surgery, and then before the first, third, and sixth cycle of chemotherapy) in order to determine serum levels of pro- and anti-inflammatory cytokines and antioxidant enzymes. RESULTS: The levels of catalase (CAT), glutathione reductase (GR), interleukin (IL)-10, IL-1α, IL-4 differed significantly depending on the stage of therapy and the type of cancer. The serum levels of IL-4 and IL-10 in patients with ovarian cancer were statistically significantly higher than those observed in patients with endometrial cancer. The activity of the tested antioxidant enzymes varied depending on the chemotherapy cycle. Their highest activity in most cases was observed before the third cycle of chemotherapy, and it decreased before the sixth cycle, irrespective of the type of cancer. CONCLUSIONS: In the studied group of patients with ovarian and endometrial cancer, the applied chemotherapy significantly changed the concentration and activity of some interleukins and antioxidant enzymes. The type of tumor determined the levels of IL-4 and IL-10 before the treatment. Evaluation of inflammatory parameters and oxidative stress in women with cancer of the reproductive organ may help to understand physiological changes resulting from the applied therapy.

Elevated serum fluoride levels in perimenopausal women are related to the components of metabolic syndrome.

OBJECTIVE: Increased fluoride levels can lead to numerous complications, including skeletal effects, cardiotoxicity, endocrine dysfunction, neurotoxicity, hepatotoxicity and nephrotoxicity. The aim of this study was to analyze the relationship between serum fluoride levels and MetS or its individual components, and to assess the diagnostic usefulness of fluoride as a factor contributing to MetS. PATIENTS AND METHODS: The study included a group of 475 women (mean age of 52.9 years), living in the West Pomeranian Voivodeship in Poland. The study involved data collection and biochemical analysis. RESULTS: Analysis of the relationship between the levels of fluoride and the presence of MetS or its components showed that the mean fluoride level was statistically significantly higher in patients with hypertriglyceridemia (dCohen = 0.39; 95% CI; confidence limits: 0.13, 0.63) and hypertension (dCohen = 0.25; 95% CI; confidence limits: 0.07, 0.44). Moreover, the mean fluoride level was significantly higher in women who met the diagnostic criteria for MetS than in the remaining subjects (dCohen = 0.40; 95% CI; confidence limits: 0.17, 0.64). CONCLUSIONS: Elevated serum fluoride levels may be associated with an increased incidence of MetS among perimenopasal women, although its diagnostic value as a marker of MetS is limited.

Gut microbiota and its metabolic potential.

OBJECTIVE: The incidence of obesity and other metabolic-related diseases has been gradually increasing. Multiple genetic as well as environmental factors play a significant role in the pathogenesis of these entities. Currently, the involvement of gut microbiota in metabolic processes has been acknowledged. This paper focuses on obesity, type 2 diabetes, and nonalcoholic fatty liver disease regarding their link with microbiome structure and its function. MATERIALS AND METHODS: We analyzed literature available in PubMed, Embase, and Google Scholar databases regarding a linkage of metabolic-associated diseases and gut microbiota RESULTS: Gut microbiota plays a significant role in host metabolism. Depending on its composition; however, it may contribute to the development of metabolic-associated diseases. In this context, not only composition of gut microbiota is important, but also its activity. Short-chain fatty acids or lipopolysaccharides are crucial metabolites involved in maintaining metabolic balance. CONCLUSIONS: Gut microbiota malfunctions might potentially induce obesity, type 2 diabetes, and nonalcoholic fatty liver disease.

Conjugated linoleic acids diminish glycogen synthase and glycogen synthase kinase-3 expression in muscle cells of C57BL/6J mice - in vitro and in vivo study.

Conjugated linoleic acids (CLA) have been extensively advertised as dietary supplements to reduce fat and increase muscle mass. However, the role of CLA in glycogen metabolism is still largely unknown. The aim of this study was to assess the effect of CLA on glycogen synthesis in vitro (CCL 136 cell line human) and CLA in vivo (C57BL/6J mice). The materials used were the CCL 136 muscle cell line and muscles of female C57BL/6J mice (n = 52), housed at animal laboratory facility and feed with "MURIGRAN", a standard feed prepared for rodents (Agropol, Poland). Chemically pure fatty acids were added to soybean oil. CLA isomers (c9,t11 CLA, t10,c12 CLA, and as a mixture (1:1)) were administered with feed. Supplementation in mice started at week 6 of age and lasted for 4 weeks. Methods used in the study were real time- PCR - quantification of gene expression, Western blot glycogen synthase kinase-3 (GSK3α 9) and glycogen synthase (GS) protein, glycogen staining by PAS. Quantitative determination of glycogen by spectrophotometry and intracellular reactive oxygen species was measured the intracellular oxidation of dichloro-dihydro-fluorescein diacetate (DCFH-DA). In vitro data showed that GS and GSK3 expression was lower in cells cultured with different CLAs and a mixture of CLAs. GS gene expression was significantly decreased in cells cultured with c9, t11 CLA (P < 0.04) and t10, c12 CLA (P < 0.05) as well as the mixture of both isomers. The GSK3α gene expression was reduced in cells cultured with a mixture of CLA (P < 0.02), whereas phosphorylation of GSK3α increased in cells cultured with c9, t11 CLA GSK3α (P < 0.05). In vivo data showed a reduction in the glycogen concentration among mice fed a diet containing t10, c 12 CLA and a mixture of CLA isomers. We conclude that both CLA isomers can affect the synthesis of glycogen in muscle cells through the regulation of GS and GSK3α gene expression.

Relationship between antioxidant defense in Acanthamoeba spp. infected lungs and host immunological status.

The role of oxidative stress in the pathogenicity of acanthamoebiasis is an important aspect of the intricate and complex host-parasite relationship. The aim of this experimental study was to determine oxidative stress through the assessment of lipid peroxidation product (LPO) levels and antioxidant defense mechanism in Acanthamoeba spp. lung infections in immunocompetent and immunosuppressed hosts. In Acanthamoeba spp. infected immunocompetent mice we noted a significant increase in lung lipid peroxidation products (LPO) at 8 days and 16 days post infection (dpi). There was a significant upregulation in lung LPO in immunocompetent and immunosuppressed mice infected by Acanthamoeba spp. at 16 dpi. The superoxide dismutase activity decreased significantly in lungs in immunosuppressed mice at 8 dpi. The catalase activity was significantly upregulated in lungs in immunocompetent vs. immunosuppressed group and in immunocompetent vs. control mice at 16 dpi. The glutathione reductase activity was significantly lower in immunosuppressed group vs. immunosuppressed control at 24 dpi. We found significant glutathione peroxidase downregulation in immunocompetent and immunosuppressed groups vs. controls at 8 dpi, and in immunosuppressed vs. immunosuppressed control at 16 dpi. The consequence of the inflammatory response in immunocompetent and immunosuppressed hosts in the course of experimental Acanthamoeba spp. infection was the reduction of the antioxidant capacity of the lungs resulting from changes in the activity of antioxidant enzymes. Therefore, the imbalance between oxidant and antioxidant processes may play a major role in pathology associated with Acanthamoeba pneumonia.

Pre-and postnatal exposition to fluorides induce changes in rats liver morphology by impairment of antioxidant defense mechanisms and COX induction.

INTRODUCTION: Fluorides are common in the environment and are absorbed mostly in the stomach and gut, it can easily move through cell membranes and its accumulation can cause harmful effects in skeletal and soft tissues. One of the most important F- accumulation sites is the liver. The aim of this study was to determine whether F- can cause inflammation in rat liver by affecting the activity of antioxidant enzymes and changes in the synthesis of prostaglandin E2 (PGE2) and thromboxane B2 (TXB2). MATERIALS AND METHODS: An in vivo model of prenatal and postnatal exposure to sodium fluoride (NaF) was used to carry out the experiment. Animals from control group received tap water to drink, while animals exposed to F- received drinking water containing NaF, 50 mg/L. In serum and liver we analyzed F- concentration, in liver - antioxidant enzymes activity, PGE2 and TXB2 concentration and immunolocalization of COX1 and COX2 proteins were measured. RESULTS: We observed significant changes in F- concentration only in liver. The results of this study showed that F- affects antioxidant enzymes activity, COX2 protein expression and PGE2 synthesis in liver. Also, in some regions of the liver of rats exposed to F-, the hepatocytes were diffusely altered, with changes resembling microvesicular steatosis. CONCLUSION: Chronic exposure to F- during development causes an accumulation of this element in the liver and changes in antioxidant enzymes activity and cyclooxygenase expression. Long term exposure to this element is toxic to the liver and can cause disturbances in its homeostasis.

The short chain fatty acids and lipopolysaccharides status in Sprague-Dawley rats fed with high-fat and high-cholesterol diet.

Short chain fatty acids (SCFA) are produced by the gut microbiota during the fermentation of non-digestible polysaccharides. Diet is a major factor driving the composition and metabolism of the colonic microbiota. The aim of our study was to examine how a fat-rich and cholesterol-rich diet that, which leads to many metabolic disorders, affects the SCFA profile and lipopolysaccharide (LPS) concentration. The experiment was carried out on 72 male, 8-weeks-old Sprague-Dawley rats. The study group (n = 30 rats) received high-fat and high cholesterol diet (HFHCh). The control group (n = 30) received standard food for laboratory rats. The rats from study and control groups were sacrificed after 4, 8, 12, 16 and 20 weeks after start of dietary exposure. The analysis of SFA in feces was performed using gas chromatography (Agilent Technologies 1260 A GC). The exposure to high-fat and high-cholesterol diet was associated with significant changes in SCFA levels. Relative to the control, each of HFHCh subgroup revealed a statistically significant decrease in butyrate (12.5% ± 5.7% versus 32.8% ± 9.1%) and an increase in propionate level (45.4% ± 6.2% versus 19.14% ± 7.1%). The ratio of acetate: propionate: butyrate was also changed (from 1.1: 0.6: 1 for control groups to 3 : 3,6 : 1 for HFHCh groups). The main SCFA in the HFHCh group was propionate instead of acetate. The dietary exposure resulted in significant differences in LPS concentration. After 12 weeks of HFD exposure, LPS concentration was significantly higher compared to control groups (P < 0.05). Our study showed that HFHCh diet affected butyrate and propionate production associated with an increase in LPS secretion. The hypothesis that observed changes could result in intestinal imbalance secondary to gut barrier dysfunction requires further studies.

Levels of Antioxidant Activity and Fluoride Content in Coffee Infusions of Arabica, Robusta and Green Coffee Beans in According to their Brewing Methods.

Coffee is a rich source of dietary antioxidants, and this property links with the fact that coffee is one of the world's most popular beverages. Moreover, it is a source of macro- and microelements, including fluoride. The aim of this work was to determine antioxidant activity of coffee beverages and fluoride content depending on different coffee species and conditions of brewing. Three species of coffee, arabica, robusta and green coffee beans obtained from retail stores in Szczecin (Poland) were analyzed. Five different techniques of preparing drink were used: simple infusion, french press, espresso maker, overflow espresso and Turkish coffee. Antioxidant potential of coffee beverages was investigated spectrophotometrically by DPPH method. Fluoride concentrations were measured by potentiometric method with a fluoride ion-selective electrode. Statistical analysis was performed using Stat Soft Statistica 12.5. Antioxidant activity of infusions was high (71.97-83.21% inhibition of DPPH) depending on coffee species and beverage preparing method. It has been shown that the method of brewing arabica coffee and green coffee significantly affects the antioxidant potential of infusions. The fluoride concentration in the coffee infusions changed depending, both, on the species and conditions of brewing, too (0.013-0.502 mg/L). Methods of brewing didn't make a difference to the antioxidant potential of robusta coffee, which had also the lowest level of fluoride among studied species. Except overflow espresso, the fluoride content was the highest in beverages from green coffee. The highest fluoride content was found in Turkish coffee from green coffee beans.

The effect of low levels of lead (Pb) in the blood on levels of sphingosine-1-phosphate (S1P) and expression of S1P receptor 1 in the brain of the rat in the perinatal period.

Sphingolipids are the main components of the lipid membrane. They also perform structural functions and participate in many signal transmission processes. One of the bioactive sphingolipids is sphingosine-1-phosphate (S1P), a ligand for five G protein-coupled receptors (S1PRs1-5), which can also act as an intracellular second messenger. S1P is responsible for the stimulation of progenitor cells in the brain, but it can also induce apoptosis of mature neurons. This study is aimed at assessing the effect of pre- and neonatal exposure to permissible Pb concentrations on S1P levels and S1PR1 (EDG1) expression in the prefrontal cortex, cerebellum, and hippocampus of rats. The concentrations of S1P were determined by RP-HPLC, S1PR1 expression was determined by RT PCR and Western Blot, and receptor immunolocalization was determined by immunohistochemistry method. Our results showed that even low blood Pb concentrations, i.e. within the acceptable limit of 10 µg/dL caused changes in the concentration of S1P in the cerebellum, prefrontal cortex, and hippocampus. Our data also showed a significant decrease in the level of S1PR1 in all studied part of brain, without significant changes in S1PR1 gene expression. Pre- and neonatal exposure to Pb also resulted in a decrease in the expression of S1PR1 in glial cells in all regions of the Cornu Ammonis (CA1-CA4) and Dentate Gyrus in the hippocampus, as well as in all layers of the cerebellum and prefrontal cortex, compared to the unexposed control group.

The Influence of Fluorine on the Disturbances of Homeostasis in the Central Nervous System.

Fluorides occur naturally in the environment, the daily exposure of human organism to fluorine mainly depends on the intake of this element with drinking water and it is connected with the geographical region. In some countries, we can observe the endemic fluorosis-the damage of hard and soft tissues caused by the excessive intake of fluorine. Recent studies showed that fluorine is toxic to the central nervous system (CNS). There are several known mechanisms which lead to structural brain damage caused by the excessive intake of fluorine. This element is able to cross the blood-brain barrier, and it accumulates in neurons affecting cytological changes, cell activity and ion transport (e.g. chlorine transport). Additionally, fluorine changes the concentration of non-enzymatic advanced glycation end products (AGEs), the metabolism of neurotransmitters (influencing mainly glutamatergic neurotransmission) and the energy metabolism of neurons by the impaired glucose transporter-GLUT1. It can also change activity and lead to dysfunction of important proteins which are part of the respiratory chain. Fluorine also affects oxidative stress, glial activation and inflammation in the CNS which leads to neurodegeneration. All of those changes lead to abnormal cell differentiation and the activation of apoptosis through the changes in the expression of neural cell adhesion molecules (NCAM), glial fibrillary acidic protein (GFAP), brain-derived neurotrophic factor (BDNF) and MAP kinases. Excessive exposure to this element can cause harmful effects such as permanent damage of all brain structures, impaired learning ability, memory dysfunction and behavioural problems. This paper provides an overview of the fluoride neurotoxicity in juveniles and adults.

Beer as a Rich Source of Fluoride Delivered into the Body.

Fluoride is an element which in the minimum amount is necessary for the proper construction of the teeth and bones. But on the other hand, it increases the synthesis of reactive oxygen species, inflammatory mediators, and impairs the action of enzymes. Beer is the most popular alcoholic beverage in the world. Due to its prevalence and volume of consumption, it should be considered as a potential source of F- and taken into account in designing a balanced diet. Therefore, the aim of this study was to analyze beer samples in terms of F- levels. The concentrations of fluoride were examined using ion-selective electrode Thermo Scientific Orion and statistical analysis was based on two-way ANOVA and t test. When compared to imported beers, Polish beers were characterized by the lowest mean F- concentration (0.089 ppm). The highest mean F- concentrations were recorded in beers from Thailand (0.260 ppm), Italy (0.238 ppm), Mexico (0.210 ppm), and China (0.203 ppm). Our study shows that beer is a significant source of fluoride for humans, which is mainly associated with the quality of the water used in beer production.

The inflammatory effect of infection with Hymenolepis diminuta via the increased expression and activity of COX-1 and COX-2 in the rat jejunum and colon.

The aim of this study was to determine whether Hymenolepis diminuta may affect the expression and activity of cyclooxygenase 1 (COX-1) and cyclooxygenase 2 (COX-2), resulting in the altered levels of their main products - prostaglandins (PGE2) and thromboxane B2 (TXB2). The study used the same experimental model as in our previous studies in which we had observed changes in the transepithelial ion transport, tight junctions and in the indicators of oxidative stress, in both small and large intestines of rats infected with H. diminuta. In this paper, we investigated not only the site of immediate presence of the tapeworm (jejunum), but also a distant site (colon). Inflammation related to H. diminuta infection is associated with the increased expression and activation of cyclooxygenase (COX), enzyme responsible for the synthesis of PGE2 and TXB2, local hormones contributing to the enhanced inflammatory reaction in the jejunum and colon in the infected rats. The increased COX expression and activity is probably caused by the increased levels of free radicals and the weakening of the host's antioxidant defense induced by the presence of the parasite. Our immunohistochemical analysis showed that H. diminuta infection affected not only the intensity of the immunodetection of COX but also the enzyme protein localization within intestinal epithelial cells - from the entire cytoplasm to apical/basal regions of cells, or even to the nucleus.

Propylparaben-induced disruption of energy metabolism in human HepG2 cell line leads to increased synthesis of superoxide anions and apoptosis.

The effect of propylparaben (in final concentrations 0.4 ng/ml, 2.3 ng/ml and 4.6 ng/ml) on the energy metabolism of HepG2 hepatocytes, superoxide anion synthesis, apoptosis and necrosis is described. Propylparaben can be toxic to liver cells due to the increased production of superoxide anions, which can contribute to a reduced concentration of superoxide dismutase in vivo and impairment of the body's antioxidant mechanisms. Finally, a further reduction in the mitochondrial membrane potential and uncoupling of the respiratory chain resulting in a reduction in ATP concentration as a result of mitochondrial damage may lead to cell death by apoptosis.

Fluoride as a factor initiating and potentiating inflammation in THP1 differentiated monocytes/macrophages.

It is well known that exposure to fluorides lead to an increased ROS production and enhances the inflammatory reactions. Therefore we decided to examine whether cyclooxygenases (particular COX-2) activity and expression may be changed by fluoride in THP1 macrophages and in this way may change the prostanoids biosynthesis. In the present work we demonstrate that fluoride increased concentration of PGE2 and TXA2 in THP1 macrophages. Following exposure to 1-10 µM NaF, COX-2 protein and COX-2 transcript increased markedly. COX-2 protein up-regulation probably is mediated by ROS, produced during fluoride-induced inflammatory reactions. Additional fluoride activates the transcription factor, nuclear factor (NF)-kappaB, which is involved in the up-regulation of COX-2 gene expression. This study indicated that even in small concentrations fluoride changes the amounts and activity of COX-1 and COX-2 enzymes taking part in the initiating and development of inflammatory process.

The Fluoride Content of Yerba Mate Depending on the Country of Origin and the Conditions of the Infusion.

There are many reports of the positive effect of yerba mate on the human body. Elemental composition analysis of yerba mate revealed the presence of many microelements and macroelements, but there is no literature data referencing the content and the effect of the method of preparing the yerba mate infusion on the amount of released fluoride and thus the amount of this element supplied to the human body. Therefore, in the traditional way (cold and hot), we prepared infusions of yerba mate from different countries and determined in samples content of fluoride using potentiometric method. Hot infusions resulted in statistically significant (p = 0.03) increases in the amount of fluoride released from the dried material to the water, compared to brewing with water at room temperature. The successive refills of hot water also resulted in a release of the same amount of fluoride, although smaller than the infusion with water at room temperature (at the third refill, it was statistically significantly smaller at p = 0.003). With an increase in the number of hot water refills, the amount of fluoride released from the sample portion significantly decreased. Similar results were recorded when analyzing samples depending on the country of origin. The amount of fluoride released into the water differed statistically significantly depending on the country of origin. The most fluoride was determined in the infusions of yerba mate from Argentina and the least in infusions from Paraguay.

The content of elements in infant formulas and drinks against mineral requirements of children.

The present study aimed at analysing the content of fluorine (F), calcium (Ca), magnesium (Mg), iron (Fe) and zinc (Zn) in the drinks for children and infant formulas, a popular supplement or substitute for breast milk produced from cow milk on an industrial scale. Ca, Mg, Zn and Fe concentrations were determined using atomic absorption spectrophotometer, while F levels using a potentiometric method. F levels in the examined formula samples increased with the intended age range, until the intended age of 1 year, and then decreased. A lower content of Ca, Mg and Zn was observed in formulas intended for children <1 year of age and higher for older children. Fe content increased with the age range. A statistically significant higher content of Ca, Mg, Zn and Fe in samples intended for children with phenylketonuria in comparison to those intended for healthy children or children with food allergies was noted. The content of the analysed elements in juices and nectars showed the highest contents in products intended for infants (under 6 months of age). The lowest levels of elements tested were found in drinks for children over 6 months of age. In conclusion, the concentrations of the examined elements in infant formulas and juices for children were decidedly greater than the standards for the individual age groups. Although the absorption of these elements from artificial products is far lower than from breast milk, there is still the fear of consequences of excessive concentrations of these minerals.

The effect of reactive oxygen species on the synthesis of prostanoids from arachidonic acid.

Reactive oxygen species (ROS), such as hydrogen peroxide, superoxide anion radical or hydroxyl radical, play an important role in inflammation processes as well as in transduction of signals from receptors to interleukin -1ß (IL-1ß), tumor necrosis factor α (TNF-α) or lipopolysaccharides (LPS). NADPH oxidase increases the ROS levels, leading to inactivation of protein phosphatase 1 (PP1), protein phosphatase 2A (PP2A) and protein tyrosine phosphatase (PTP): MAPK phosphatase 1 (MKP-1). Inactivation of phosphatases results in activation of mitogen-activated protein kinase (MAPK) cascades: c-Jun N-terminal kinase (JNK), p38 and extracellular signal-regulated kinase (Erk), which, in turn, activate cytosolic phospholipase A2 (cPLA2). ROS cause cytoplasmic calcium influx by activation of phospholipase C (PLC) and phosphorylation of IP3-sensitive calcium channels. ROS activate nuclear factor κB (NF-κB) via IκB kinase (IKK) through phosphoinositide 3-kinase (PI3K), tumor suppressor phosphatase and tensin homolog (PTEN) and protein kinase B (Akt/PKB) or NF-κB-inducing kinase (NIK). IKK phosphorylates NF-κB α subunit (IκBα) at Ser³². Oxidative stress inactivates NIK and IκB kinase γ subunit/NF-κB essential modulator (IKKγ/NEMO), which might cause activation of NF-κB that is independent on IKK and inhibitor of IκBα degradation, including phosphorylation of Tyr4² at IκBα by c-Src and spleen tyrosine kinase (Syk), phosphorylation of the domain rich in proline, glutamic acid, serine and threonine (PEST) sequence by casein kinase II and inactivation of protein tyrosine phosphatase 1B (PTP1B). NF-κB and MAPK cascades-activated transcription factor activator protein 1 (AP-1) and CREB-binding protein (CBP/p300) lead to expression of cytosolic phospholipase A2 (cPLA2), cyclooxygenase-2 (COX-2) and membrane-bound prostaglandin E synthase 1 (mPGES-1), and thus to increased release of arachidonic acid and production of prostaglandins, particularly prostaglandin E2 (PGE2). ROS increase the activity of hematopoietic-type PGD synthase (H-PGDS), and, as a result, the production of prostaglandin D2 (PGD2). However, the superoxide radical reacts with nitric oxide forming peroxynitrite that inactivates prostaglandin I synthase (PGIS), suppressing the production of prostaglandin I2 (PGI2). ROS do not affect thromboxane synthesis in a direct manner; this is achieved by an increase in cPLA2 activity and COX-2 expression. The aim of this review was to summarize knowledge of influence of ROS on the synthesis of prostanoids from arachidonic acid.

Perinatal exposure to lead induces morphological, ultrastructural and molecular alterations in the hippocampus.

The aim of this paper is to examine if pre- and neonatal exposure to lead (Pb) may intensify or inhibit apoptosis or necroptosis in the developing rat brain. Pregnant experimental females received 0.1% lead acetate (PbAc) in drinking water from the first day of gestation until weaning of the offspring; the control group received distilled water. During the feeding of pups, mothers from the experimental group were still receiving PbAc. Pups were weaned at postnatal day 21 and the young rats of both groups then received only distilled water until postnatal day 28. This treatment protocol resulted in a concentration of Pb in rat offspring whole blood (Pb-B) below the threshold of 10 µg/dL, considered safe for humans.We studied Casp-3 activity and expression, AIF nuclear translocation, DNA fragmentation, as well as Bax, Bcl-2 mRNA and protein expression as well as BDNF concentration in selected structures of the rat brain: forebrain cortex (FC), cerebellum (C) and hippocampus (H). The microscopic examinations showed alterations in hippocampal neurons.Our data shows that pre- and neonatal exposure of rats to Pb, leading to Pb-B below 10 µg/dL, can decrease the number of hippocampus neurons, occurring concomitantly with ultrastructural alterations in this region. We observed no morphological or molecular features of severe apoptosis or necrosis (no active Casp-3 and AIF translocation to nucleus) in young brains, despite the reduced levels of BDNF. The potential protective factor against apoptosis was probably the decreased Bax/Bcl-2 ratio, which requires further investigation. Our findings contribute to further understanding of the mechanisms underlying Pb neurotoxicity and cognition impairment in a Pb-exposed developing brain.

Fluoride in low concentration modifies expression and activity of 15 lipoxygenase in human PBMC differentiated monocyte/macrophage.

Epidemiological and experimental evidences demonstrate positive correlation between environmental and occupational fluoride exposure and risk to various cardio-respiratory disorders. That fore we decided to examine the effect of fluorides on activity and expression of 15LOX enzyme which is implicated in biosynthesis of inflammatory mediators. Expression of 15LOX-1 and -2 enzymes mRNA and protein was analyzed using RT PCT and immunoblotting methods respectively whereas HPLC method was used to measure the levels of 15 lipoxygenases end products. Additionally AA and LA concentration in cells was measured using GC method. We observed that fluoride in small concentration may significantly decrease activity of 15LOX-1 and -2 in human PBMC macrophages and then concentration of its end products: 15-HETE, 12-HETE and 9+13-HODE, what may cause development of inflammation through the cholesterol arrest into the macrophages and its differentiation to foam cell. Noted by our team overexpression of the 15LOX-1 enzyme in macrophages after addition of lowest fluoride concentrations (1 and 3 µM) may be aimed at fighting inflammation development and excessive intracellular lipid accumulation. But highest fluoride concentrations (6 and 10 µM) added to cell culture slowly declined expression of this enzyme probably because of developing inflammation. Additional 15LOX-2 expression in macrophages after fluoride addition was low in 1 and 3 µM concentrations, but increased significantly after 10 µM fluoride addition what may suggest developing acute inflammation, because 15LOX-2 is associated to increased local hypoxia. This study indicated that even in small concentrations fluorides changes the amounts and activity of 15 LOX-1 and -2 enzymes taking part in the development of inflammatory process.

Activation of phospholipase A(2) by low levels of fluoride in THP1 macrophages via altered Ca(2+) and cAMP concentration.

Phospholipases (PLA's) participate in the regulation of physiological and pathological processes in the cell, including the release of pro-inflammatory mediators and stimulation of inflammatory processes. It is also well known that fluoride can increase the inflammatory reactions. Therefore we decided to examine the effect of fluorides in concentrations determined in human serum on cPLA(2) and sPLA(2) activity. The incubation of macrophages in fluoride solutions significantly increased the amount of synthesized cellular cAMP, intracellular calcium and sPLA(2) activity in a dose-dependent pattern. The cPLA(2) activity, estimated by the amount of released arachidonic acid, increased significantly when 10 µM NaF was used. The results of our study suggest that fluoride may change the activity of phospholipases in macrophage cells. Probably, increased cAMP concentration activates protein kinase C (PKC) and thus stimulates PLA(2). cAMP also regulates the passage of Ca(2+) through ion channels, which additionally influence PLA(2) throughout Ca(2+)-calmodulin dependent protein kinase.