Blastocystis spp. and Other Intestinal Parasites in Polish Soldiers Deployed to Lebanon and Iraq.

Intestinal parasitic infections are one of the most common infectious diseases worldwide, particularly in developing countries. A distinct group at increased risk of infection is military personnel deployed overseas for extended periods, typically six months at a time. The aim of this study was to determine the prevalence of Blastocystis spp. and other intestinal parasites in Polish military personnel returning from deployments to Lebanon (n = 206) and Iraq (n = 220). In this group of subjects, we found Blastocystis spp. (13.6%), Dientamoeba fragilis (3.3%), Entamoeba coli (0.9%), and Endolimax nana (0.5%). Entamoeba histolytica sensu lato and Chilomastix mesnili infections were detected only in one soldier returning from Lebanon and Iraq, respectively. Blastocystis subtype (ST) 3 was predominant in soldiers returning from Lebanon, followed by ST2 and ST1. ST1 infection was predominant in soldiers returning from Iraq, followed by ST3 and ST2. Our study affirms that, deployment abroad is of no influence of the prevalence of parasitic protozoa. However, it would be worth to monitor parasite infection in military personnel returning from tropical zone even if they have no actual symptoms. In addition, it is very important to determine the subtypes of Blastocystis-this may help to clearly define their pathogenicity, especially considering the scarcity of studies on Blastocystis genotypes in Iraqi and Lebanese residents.

The role of apoptosis and oxidative stress in the pathophysiology of Acanthamoeba spp. infection in the kidneys of hosts with different immunological status.

BACKGROUND: Acanthamoeba spp. are opportunistic pathogens that cause inflammation, mostly in the brain, lungs and cornea. Recent reports indicate kidney dysfunction in hosts with systemic acanthamoebiasis. The aim of the study was to analyze the gene expression and protein concentration of NADPH oxidase 2 and 4 (NOX2 and NOX4, respectively) and nuclear erythroid 2-related factor (Nrf2) in the kidneys of hosts with systemic acanthamoebiasis. We also aimed to determine the protein and gene expressions of Bcl2, Bax, caspases 3 and 9. METHODS:  Mice were divided into four groups based on their immunological status and Acanthamoeba sp. infection: A, immunocompetent Acanthamoeba sp.-infected mice; AS, immunosuppressed Acanthamoeba sp.- infected mice; C, immunocompetent uninfected mice; CS, immunosuppressed uninfected mice. NOX2, NOX4 and Nrf2 were analyzed by quantitative reverse transcription PCR (qRT-PCR) and ELISA methods, while pro-apoptotic and anti-apoptotic proteins (Bax and Bcl-2, respectively), Cas9, Cas3 were analyzed by qRT-PCR and western blot methods.  RESULTS: Increased gene expression and/or protein concentration of NOX2 and NOX4 were found in both immunocompetent and immunosuppressed mice infected with Acanthamoeba sp. (groups A and AS, respectively). Gene expression and/or protein concentration of Nrf2 were higher in group A than in control animals. Compared to control mice, in the AS group the expression of the Nrf2 gene was upregulated while the concentration of Nrf2 protein was decreased. Additionally in A group, higher gene and protein expression of Bcl-2, and lower gene as well as protein expression of Bax, caspases 3 and 9 were noted. In contrast, the AS group showed lower gene and protein expression of Bcl-2, and higher gene as well as protein expression of Bax, caspases 3 and 9. CONCLUSIONS: This study is the first to address the mechanisms occurring in the kidneys of hosts infected with Acanthamoeba sp. The contact of Acanthamoeba sp. with the host cell surface and/or the oxidative burst caused by elevated levels of NOXs lead to an antioxidant response enhanced by the Nrf2 pathway. Acanthamoeba sp. have various strategies concerning apoptosis. In immunocompetent hosts, amoebae inhibit the apoptosis of kidney cells, and in immunosuppressed hosts, they lead to increased apoptosis by the intrinsic pathway and thus to a more severe course of the disease.

Oxidative Stress in Pregnancy.

Recent years have seen an increased interest in the role of oxidative stress (OS) in pregnancy. Pregnancy inherently heightens susceptibility to OS, a condition fueled by a systemic inflammatory response that culminates in an elevated presence of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in the circulatory system. The amplified OS in pregnancy can trigger a series of detrimental outcomes such as underdevelopment, abnormal placental function, and a host of pregnancy complications, including pre-eclampsia, embryonic resorption, recurrent pregnancy loss, fetal developmental anomalies, intrauterine growth restriction, and, in extreme instances, fetal death. The body's response to mitigate the uncontrolled increase in RNS/ROS levels requires trace elements that take part in non-enzymatic and enzymatic defense processes, namely, copper (Cu), zinc (Zn), manganese (Mn), and selenium (Se). Determination of ROS concentrations poses a challenge due to their short half-lives, prompting the use of marker proteins, including malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), catalase (CAT), and glutathione (GSH). These markers, indicative of oxidative stress intensity, can offer indirect assessments of pregnancy complications. Given the limitations of conducting experimental studies on pregnant women, animal models serve as valuable substitutes for in-depth research. This review of such models delves into the mechanism of OS in pregnancy and underscores the pivotal role of OS markers in their evaluation.

Demodex Species and Culturable Microorganism Co-Infestations in Patients with Blepharitis.

We aimed to determine the prevalence of Demodex spp. and bacterial infection in patients with blepharitis and also to investigate the relationship between culturable microorganisms and Demodex spp. in this study. The study included patients diagnosed with blepharitis (n = 128) and volunteers without ocular problems (n = 113). Eyelash sampling was performed by epilating eight lashes, which were then tested for Demodex spp. using a light microscope. The examination consisted of assessing the patient's vision with and without ocular correction and tonus in both eyes and a careful examination of the anterior segment of both eyes. Bacterial identification was performed based on morphological, physiological, and biochemical methods. The prevalence of Demodex spp. was 8.0% in patients from the control group and all patients with blepharitis. Isolated forms of Demodex spp. were detected in all infested patients in the control group and in 58% of patients with blepharitis. A total of 35% of patients with blepharitis had from three to nine forms of Demodex spp., and 7% of patients with blepharitis had more than 10 mites in every field of vision. We found a statistically significant relationship between Demodex spp. infestation and the occurrence of eye dryness and sensations of burning and tearing, redness of the conjunctiva, feeling of a foreign body, loss of eyelashes, Meibomian gland dysfunction, and cylindrical dandruff. There were statistically significant relationships between Demodex sp. infestation and the presence of hyperopia, Meibomian cysts, chronic eyelid inflammation, and the use of eyeglasses. There was also a statistically significant relationship between the occurrence of Demodex spp. and seborrheic dermatitis and diabetes mellitus. Culturable microorganisms of the ocular surface were found in 8.7% of participants who were uninfested and in all patients infested with D. folliculorum. We isolated Staphylococcus aureus, Acinetobacter baumannii, Streptococcus pneumoniae, Klebsiella oxytoca, and Bacillus spp. from the conjunctival sac only in patients infested with D. folliculorum. This indicates an increased probability of colonization by pathogenic bacteria in patients with demodicosis. Therefore, patients infested with D. folliculorum should undergo a microbiological examination of conjunctival swabs.

Macroelement and Microelement Levels in the Urine in Experimental Acanthamoebiasis.

Free-living amoebas can impact the excretion of macroelements and microelements in urine. The aim of the present study was to examine the concentrations of macroelements, including calcium (Ca), phosphorus (P), sodium (Na), potassium (K), and magnesium (Mg), as well as microelements such as manganese (Mn), zinc (Zn), copper (Cu), iron (Fe), and chromium (Cr), in the urine during acanthamoebiasis while considering the host's immunological status. This is the first study to show an increase in urinary excretion of Ca, Mn, Cu, Fe, Na, and Cr, along with a decreased excretion of K, in immunocompetent mice 16 days post Acanthamoeba sp. infection. In the final phase of infection (24 dpi), there was a further decrease in urinary K excretion and a lower level of P in Acanthamoeba sp. infected immunocompetent hosts. During acanthamoebiasis in immunosuppressed hosts, increased excretion of Zn, Fe, and Cr was observed at the beginning of the infection, and increased Na excretion only at 16 days post Acanthamoeba sp. infection. Additionally, host immunosuppression affected the concentration of Fe, Cr, Zn, Cu, Mn, and Ca in urine.

The Immunological Changes in the Skin of BALC/c Mice with Disseminated Acanthamoebiasis.

Toll-like receptors (TLR) are involved in the recognition of numerous pathogens, including Acanthamoeba spp. Thanks to this, it is possible for immune cells to recognize microorganisms and trigger the body's innate immune response. The stimulation of TLRs also leads to the activation of specific immunity. The aim of the study was to determine the TLR2 and TLR4 gene expression in the skin of BALC/c mice infected with Acanthamoeba with AM22 strain isolated from a patient. Receptor expression was assessed by real-time polymerase chain reaction (qPCR) in the amoeba-infected host with normal (A) and reduced immunity (AS) as well as in the control host with normal immunity (C) and reduced immunity (CS). Statistical analysis of TLR2 gene expression in A and AS groups compared to C and CS groups, respectively, were statistically insignificant. In the A group, we found statistical upregulation of TLR4 gene expression at 8 dpi compared to the C group. While in AS group, TLR4 gene expression was at a similar level, such as in the CS group. Taking into account the host's immune status, the TLR4 gene expression was statistically higher in the skin of host from A group than in host from AS group at the beginning of the infection. Increased TLR4 gene expression in hosts with normal immunity infected with Acanthamoeba suggests the involvement of the studied receptor in the course of acanthamoebiasis. The above research results provide new data on the involvement of the studied receptor in the skin in the host's immune defense triggered during the Acanthamoeba infection.

The Trace Element Concentrations and Oxidative Stress Parameters in Afterbirths from Women with Multiple Pregnancies.

The aim of this study was to evaluate the intensity of oxidative stress by measuring the concentrations of lipid peroxidation products (LPO) in fetal membrane, umbilical cord, and placenta samples obtained from women with multiple pregnancies. Additionally, the effectiveness of protection against oxidative stress was assessed by measuring the activity of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione reductase (GR). Due to the role of iron (Fe), copper (Cu), and zinc (Zn) as cofactors for antioxidant enzymes, the concentrations of these elements were also analyzed in the studied afterbirths. The obtained data were compared with newborn parameters, selected environmental factors, and the health status of women during pregnancy to determine the relationship between oxidative stress and the health of women and their offspring during pregnancy. The study involved women (n = 22) with multiple pregnancies and their newborns (n = 45). The Fe, Zn, and Cu levels in the placenta, umbilical cord, and fetal membrane were determined using inductively coupled plasma atomic emission spectroscopy (ICP-OES) using an ICAP 7400 Duo system. Commercial assays were used to determine SOD, GPx, GR, CAT, and LPO activity levels. The determinations were made spectrophotometrically. The present study also investigated the relationships between trace element concentrations in fetal membrane, placenta, and umbilical cord samples and various maternal and infant parameters in women. Notably, a strong positive correlation was observed between Cu and Zn concentrations in the fetal membrane (p = 0.66) and between Zn and Fe concentrations in the placenta (p = 0.61). The fetal membrane Zn concentration exhibited a negative correlation with shoulder width (p = -0.35), while the placenta Cu concentration was positively correlated with placenta weight (p = 0.46) and shoulder width (p = 0.36). The umbilical cord Cu level was positively correlated with head circumference (p = 0.36) and birth weight (p = 0.35), while the placenta Fe concentration was positively correlated with placenta weight (p = 0.33). Furthermore, correlations were determined between the parameters of antioxidative stress (GPx, GR, CAT, SOD) and oxidative stress (LPO) and the parameters of infants and maternal characteristics. A negative correlation was observed between Fe and LPO product concentrations in the fetal membrane (p = -0.50) and placenta (p = -0.58), while the Cu concentration positively correlated with SOD activity in the umbilical cord (p = 0.55). Given that multiple pregnancies are associated with various complications, such as preterm birth, gestational hypertension, gestational diabetes, and placental and umbilical cord abnormalities, research in this area is crucial for preventing obstetric failures. Our results could serve as comparative data for future studies. However, we advise caution when interpreting our results, despite achieving statistical significance.

Red fox as a potential bioindicator of metal contamination in a European environment.

In times of widespread environmental pollution with heavy metals of anthropogenic origin and the increasing dynamics of this process, it is justified to collect as much data as possible on the concentration of metals in terrestial mammals from unpolluted areas. The purpose of this research was to present the concentration of essential (chromium, copper, iron, manganese, molybdenum, and zinc), probably essential (nickel, vanadium) and non-esential element (cadmium, lead, silver, strontium, and tin) in the liver, kidneys, muscles and brain of red fox (Vulpes vulpes) inhabiting north-western Poland. We revealed that the concentration of all metals, apart from Ni, was different between studied tissues. Sn and Mo have the highest affinity to the liver, whereas Ag, Sr, Cr had higher concentrations in the brain than in other organs. Various positive relationship between the concentrations of metals were observed in the tissues. Moreover, we noted negative correlations between Ag and Sn in the kidneys and brain, and between Cu and Ag in the liver. In our study the red fox was used as biomonitor for the assessment of exposure of carnivores to metals, in the generally unpolluted areas of Central Europe. Data presented in the report may be used as comparative values in similar ecotoxicological studies.

Protozoan Intestinal Parasitic Infection in Patients with Hematological Malignancies.

The aim of this study was to evaluate the frequency of gastrointestinal protozoan infection in patients with hematological malignancies (HMs) undergoing intensive hemato-oncological treatment and to determine the influence of certain biological factors on the incidence of intestinal parasite infection. Stool samples were collected from hematological malignancy patients (n = 50) hospitalized at the Department of Hematology and Transplantology of the Pomeranian Medical University in Szczecin. The control group consisted of 50 healthy participants. We used a direct smear examination and a commercial immunoenzymatic test. Intestinal protozoans were detected in 16% of patients with hematological malignancies and in 6% of individuals in the control group. In stool samples from patients with HM, cysts of Giardia intestinalis (2%), oocysts of Cryptosporidium spp. (10%), vacuolar forms of potentially pathogenic Blastocystis spp. (2%), and cysts of nonpathogenic Entamoeba coli (2%) were found. Cryptosporidium spp. and Giardia intestinalis coproantigens were detected in 5 (10%) and 1 (2%) patients with HM, respectively. In three participants from the control group, vacuolar forms of Blastocystis spp. were found. In the patients with HM, a significantly higher prevalence of intestinal parasite infection was found in individuals working in the garden without protective gloves and those in contact with animals. In patients with hematological malignancies, intestinal parasites should be excluded, even during intensive chemotherapy treatment.

Neurotrophic Factors in Experimental Cerebral Acanthamoebiasis.

To date, no studies have addressed the role of neurotrophins (NTs) in Acanthamoeba spp. infections in the brain. Thus, to clarify the role of NTs in the cerebral cortex and hippocampus during experimental acanthamoebiasis in relation to the host immune status, the purpose of this study was to determine whether Acanthamoeba spp. may affect the concentration of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4) in brain structures. Our results suggest that at the beginning of infection in immunocompetent hosts, BDNF and NT-3 may reflect an endogenous attempt at neuroprotection against Acanthamoeba spp. infection. We also observed a pro-inflammatory effect of NGF during acanthamoebiasis in immunosuppressed hosts. This may provide important information for understanding the development of cerebral acanthamoebiasis related to the immunological status of the host. However, the pathogenesis of brain acanthamoebiasis is still poorly understood and documented and, therefore, requires further research.

Concentrations of PGE2 and TXB2 in the Eyes of Mice with Disseminated Acanthamoebiasis.

Previous studies have shown that Acanthamoeba spp. may invade the eyes by migrating along the optic nerve to the eyes from the brain. This study aimed to confirm the presence of inflammation in the eyes of mice with disseminated acanthamoebiasis by examining prostaglandin E2 (PGE2) and thromboxane B2 (TXB2) concentrations in the eyes of immunocompetent and immunocompromised mice intranasally inoculated with Acanthamoeba spp. The PGE2 concentration was statistically significantly lower in the immunocompromised amoebae-infected mice on 8 dpi compared with the noninfected group of animals, and it was higher in the eyes of immunosuppressed amoebae-infected mice on 16 dpi than in the control group of animals. There was a statistically significant lower TXB2 concentration in the eyes of immunocompetent infected mice compared with the noninfected group on 8 dpi. However, on 24 dpi, we noted statistically significant higher TXB2 levels in the immunocompetent infected mice than in the control group. In immunocompromised mice, there was a lower TXB2 level on 8 dpi than in control mice. This study confirmed the existence of an inflammatory process in the eyes of immunocompetent and immunocompromised mice infected with Acanthamoeba spp. without damaged corneas.

Selected Metal Concentration in Maternal and Cord Blood.

Essential and non-essential elements deficiencies may lead to various birth complications. The aim of this paper was to determine calcium (Ca), copper (Cu), iron (Fe), potassium (K), magnesium (Mg), sodium (Na), phosphorus (P), lead (Pb), strontium (Sr), and zinc (Zn) concentrations in maternal blood and cord blood. Whole blood and cord blood samples collected from pregnant women (n = 136) were analyzed for the concentration of the elements by spectrophotometric atomic absorption in inductively coupled argon plasma (ICP-OES). The results showed that Ca, Pb, and Sr concentrations were similar in maternal and cord blood, while Fe and K levels were higher in cord blood than in maternal blood. The cord blood Cu, Na, and Zn concentrations were lower than those in maternal blood, suggesting transplacental transfer of these elements were limited. Moreover, checking the influence of studied elements on the anthropometric parameters of the newborns, we found that the highest number of associations was between Cu in cord blood. Due to the fact that the pregnant women were healthy, and the newborns were without any disorders, we suggest that the values obtained in our study are normal values of studied elements in whole blood and cord blood in patients from Poland.

Prevalence, subtypes and risk factors of Blastocystis spp. infection among pre- and perimenopausal women.

BACKGROUND: Blastocystis spp. are considered pathogenic or commensal organisms, although the majority of researchers suggest that these are neglected pathogens. The main aim of this study was to determine the prevalence and subtype distribution of Blastocystis spp. in pre- and perimenopausal women, with respect to socio-demographic (age and place of residence), and epidemiological factors, as well as drinking tap water, contact with domestic animals, traveling abroad, health status, and presence of gastrointestinal symptoms. Additionally, the objective was to compare hematological and biochemical parameters of Blastocystis spp. infected and uninfected women. METHODS: The study included 425 women aged 45-60. Their stool samples were examined microscopically and analyzed by a conventional polymerase chain reaction (cPCR). RESULTS: Blastocystis spp. were detected in 6.1% of pre- and perimenopausal women. Molecular analysis of the stool samples identified seven Blastocystis subtypes (ST1-ST4, ST6, ST7, and ST9). Blastocystis subtypes 2 and 3 were the most prevalent. The presence of Blastocystis spp. was not significantly related to socio-demographic and epidemiological factors. There were also no significant associations between Blastocystis spp. and blood parameters, or gastrointestinal symptoms. CONCLUSION: This study complements the limited available data on the prevalence of Blastocystis spp. in pre- and perimenopausal women. It is also the first report showing the presence of Blastocystis subtype 9 in Poland.

Potential Biomarkers in Diagnosis of Renal Acanthamoebiasis.

Recent studies indicate that Acanthamoeba spp. may play a significant role in kidney dysfunction. The aim of the study was to examine the levels of kidney injury molecule 1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), and monocyte chemotactic protein 1 (MCP-1), as well as an activity of matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9, respectively) in the kidneys of immunocompetent and immunosuppressed mice infected with Acanthamoeba spp. The levels of KIM-1, NGAL, and MCP-1 were analyzed by enzyme-linked immunosorbent assay (ELISA), and the activity of MMPs was determined by gelatin zymography. The elevated KIM-1 level was found in the kidneys of immunocompetent mice at the beginning of Acanthamoeba spp. infection. In the immunosuppressed mice, the KIM-1 level was statistically different. The statistically decreased NGAL level was found in the kidneys of immunocompetent mice compared to the uninfected mice. In the immunocompromised mice, we found statistically significant differences in MCP-1 levels between the uninfected and infected groups. There was an increase in the expression of both MMP-2 and MMP-9 in the kidneys of immunocompetent and immunosuppressed mice infected with Acanthamoeba spp. compared to the uninfected mice. The results indicate that KIM-1, NGAL, MCP-1, MMP-2, MMP-9, and MMP-9/NGAL might be promising biomarkers of renal acanthamoebiasis.

Relationship between concentrations of elements and geographic location in Poland.

INTRODUCTION: Interactions occur between concentrations of metals in the human body and the environment in various geographic locations, which can be of importance for both the proper development and the course of pregnancy. OBJECTIVE: The aim of the study was to assess the concentrations of Zn, Cu and Cd and Zn:Cu and Zn:Cd molar ratios in the placenta, umbilical cord and in the foetal membrane, and to examine the relationship between concentrations of these elements and the place of residence. MATERIAL AND METHODS: The research material was obtained from 99 healthy women from north-western and central Poland. Data for the study were collected from the medical history and documents of admission to the ward and documents confirming the birth of the newborn. Concentrations of zinc (Zn), copper (Cu) and cadmium (Cd) were determined using the ICP-AES method (spectrophotometry of atomic absorption) in inductively coupled argon plasma. RESULTS: The average concentration of Zn, Cu and Cd in afterbirths was ~ 0.01 mg/kg of dry weight (dw). In central Poland, the results revealed a significant increase in zinc in the membrane (OR=1.098, p=0.002), cadmium in the placenta (OR=1.324, p=0.006), Zn:Cu in the membrane (OR=1.012, p<0.001). In north-western Poland, an increase in copper in the membrane (OR=1.239, p=0.025) was revealed. CONCLUSIONS: The use biological materials, such as the placenta, foetal membrane and umbilical cord to assess exposure to heavy metals and necessary elements is justified.

The Effects of Calcium, Magnesium, Phosphorus, Fluoride, and Lead on Bone Tissue.

Bones are metabolically active organs. Their reconstruction is crucial for the proper functioning of the skeletal system during bone growth and remodeling, fracture healing, and maintaining calcium-phosphorus homeostasis. The bone metabolism and tissue properties are influenced by trace elements that may act either indirectly through the regulation of macromineral metabolism, or directly by affecting osteoblast and osteoclast proliferation or activity, or through becoming part of the bone mineral matrix. This study analyzes the skeletal impact of macroelements (calcium, magnesium, phosphorus), microelements (fluorine), and heavy metals (lead), and discusses the concentration of each of these elements in the various bone tissues.

Pathomechanisms in the Kidneys in Selected Protozoan Parasitic Infections.

Leishmaniasis, malaria, toxoplasmosis, and acanthamoebiasis are protozoan parasitic infections. They remain important contributors to the development of kidney disease, which is associated with increased patients' morbidity and mortality. Kidney injury mechanisms are not fully understood in protozoan parasitic diseases, bringing major difficulties to specific therapeutic interventions. The aim of this review is to present the biochemical and molecular mechanisms in kidneys infected with Leishmania spp., Plasmodium spp., Toxoplasma gondii, and Acanthamoeba spp. We present available mechanisms of an immune response, oxidative stress, apoptosis process, hypoxia, biomarkers of renal injury in the serum or urine, and the histopathological changes of kidneys infected with the selected parasites. Pathomechanisms of Leishmania spp. and Plasmodium spp. infections have been deeply investigated, while Toxoplasma gondii and Acanthamoeba spp. infections in the kidneys are not well known yet. Deeper knowledge of kidney involvement in leishmaniasis and malaria by presenting their mechanisms provides insight into how to create novel and effective treatments. Additionally, the presented work shows gaps in the pathophysiology of renal toxoplasmosis and acanthamoebiasis, which need further research.

Current and historical nephric and hepatic mercury concentrations in terrestrial mammals in Poland and other European countries.

The long-term anthropogenic release of mercury (Hg) into the environment has led to contamination of the biosphere, with all forms of Hg showing toxic effects and the ability to accumulate in organisms. Since the 1970s, efforts have been made in Western Europe to reduce Hg emissions and for the economic use of Hg, leading to a reduction in Hg exposure to humans and entire ecosystems. The purpose of this research was to present the total mercury (THg) burden in three mustelids (the piscivorous Eurasian otter and American mink, and the invertebrativorous European badger) inhabiting north-western Poland (mostly floodplains) and other European countries (literature data). Moreover, we wanted to investigate whether reductions in the environmental Hg burden in Europe have resulted in reductions in liver and kidney levels in wild terrestrial mammals (Eurasian otter, wild boar, red deer, roe deer, cervids, leporids, rodents, and ecotrophic groups: piscivorous mustelids, non-mustelids whose diets include aquatic prey, canids and other carnivores, omnivores, herbivores), between samples collected before and after 2000. We revealed significantly higher nephric THg levels in roadkilled than in trapped American minks. As roadkilled piscivorous mustelids from the same floodplain had similar hepatic and nephric THg concentrations, we suggest that the European research on Hg ecotoxicology should more often use alien American mink instead of the protected Eurasian otter. Badgers inhabiting Polish and other European floodplains bioaccumulated higher amounts of THg than those from other areas, and as such, may be recommended as bioindicator of mercury soil contamination. Our analysis of abundant data on mammalian hepatic and nephric THg concentrations (excluding non-piscivores mustelids) showed that in 12 of 21 cases, Hg concentrations had dropped significantly since 2000. This data signals a reduction in Hg contamination in terrestrial mammals, such as the Eurasian otter, and may be reason for cautious optimism.

Immunopathogenicity of Acanthamoeba spp. in the Brain and Lungs.

Free-living amoebas, including Acanthamoeba spp., are widely distributed in soil, water, and air. They are capable of causing granulomatous amebic encephalitis, Acanthamoeba pneumonia, Acanthamoeba keratitis, and disseminated acanthamoebiasis. Despite low occurrence worldwide, the mortality rate of Acanthamoeba spp. infections is very high, especially in immunosuppressed hosts. Acanthamoeba infections are a medical problem, owing to limited improvement in diagnostics and treatment, which is associated with incomplete knowledge of pathophysiology, pathogenesis, and the host immune response against Acanthamoeba spp. infection. The aim of this review is to present the biochemical and molecular mechanisms of Acanthamoeba spp.-host interactions, including the expression of Toll-like receptors, mechanisms of an immune response, the activity of metalloproteinases, the secretion of antioxidant enzymes, and the expression and activity of cyclooxygenases. We show the relationship between Acanthamoeba spp. and the host at the cellular level and host defense reactions that lead to changes in the selected host's organs.

The expression of TLR2 and TLR4 in the kidneys and heart of mice infected with Acanthamoeba spp.

BACKGROUND: Acanthamoeba spp. are cosmopolitan protozoans that cause infections in the brain, as well as extracerebral infections in the cornea, lungs and skin. Little is known about the mechanisms of the immunological response to these parasites in organs which are not their main biotope. Therefore, the purpose of this study was to determine the expression of TLR2 and TLR4 in the kidneys and heart of Acanthamoeba spp.-infected mice, with respect to the host's immunological status. METHODS: The mice were grouped into four groups: immunocompetent control mice; immunosuppressed control mice; immunocompetent Acanthamoeba spp.-infected mice; and immunosuppressed Acanthamoeba spp. infected mice. In the study, we used the amoebae T16 genotype which was isolated from a patient. The TLRs expressions in the kidneys and heart of mice were assessed by quantitative real-time polymerase chain reaction. Moreover, we visualized TLR2 and TLR4 proteins in the organs by immunohistochemical staining. RESULTS: In the kidneys, we observed a higher TLR2 expression in immunosuppressed mice at 24 days post-Acanthamoeba spp. infection (dpi) compared to the uninfected mice. There were no statistically significant differences in TLR4 expression in the kidneys between the immunocompetent and immunosuppressed mice, both of infected and uninfected mice. In the heart, we observed a difference in TLR2 expression in immunocompetent mice at 24 dpi compared to immunocompetent mice at 8 dpi. The immunocompetent Acanthamoeba spp.-infected mice had higher TLR4 expression at 8 dpi compared to the immunocompetent uninfected mice. CONCLUSIONS: Our results indicate that TLR2 is involved in response to Acanthamoeba spp. infection in the kidneys, whereas in the heart, both studied TLRs are involved.