Oxidative Stress, Inflammation and Colorectal Cancer: An Overview.

Colorectal cancer (CRC) represents the second leading cause of cancer-related deaths worldwide. The pathogenesis of CRC is a complex multistep process. Among other factors, inflammation and oxidative stress (OS) have been reported to be involved in the initiation and development of CRC. Although OS plays a vital part in the life of all organisms, its long-term effects on the human body may be involved in the development of different chronic diseases, including cancer diseases. Chronic OS can lead to the oxidation of biomolecules (nucleic acids, lipids and proteins) or the activation of inflammatory signaling pathways, resulting in the activation of several transcription factors or the dysregulation of gene and protein expression followed by tumor initiation or cancer cell survival. In addition, it is well known that chronic intestinal diseases such as inflammatory bowel disease (IBD) are associated with an increased risk of cancer, and a link between OS and IBD initiation and progression has been reported. This review focuses on the role of oxidative stress as a causative agent of inflammation in colorectal cancer.

The survey of soil-transmitted helminth species abundance in Slovakia with an emphasis on parameters important for their distribution.

Soil-transmitted helminths (STH) can be easily dispensable in socially disadvantaged groups. The Roma people represent the group most at risk in Slovakia. This study aimed to investigate the occurrence of STH infections in minorities living with animals under low hygienic conditions and on contaminated soil. Subsequently, we identified the risk assessment factors of the STH transmissions based on parasitological results. In our study, STHs were predominantly found among the Roma communities living in unsanitary conditions. The prevalence of Ascaris lumbricoides among the majority was 0.79%, and Trichuris trichiura was 0.05 %. On the contrary, a community-based cross-sectional survey across eastern Slovakia also found a prevalence of 22.28 and 3.47% for A. lumbricoides and T. trichiura among the Roma population. Inhabitants that belong to the Roma minority had a 37.12 infection OR times higher than non-Roma inhabitants. The Roma people living in the countryside have a 2.23-fold higher chance of getting infected with STH than Roma living in the city. Therefore, soil and domestic animals were also examined for the presence of the STH eggs to show the environmental burden. In general, the presence of STH eggs was confirmed in 26.26% of the soil examined samples. The detailed description is as follows: eggs of Toxocara spp., Trichuris spp., eggs from the family Ancylostomatidae, and Toxascaris leonina. Ascaris eggs were detected only in the soil from localities with low hygienic standards. The probability of contracting the STH eggs in segregated settlements was 15.94 times higher compared to urban or rural areas. In addition to humans, dogs can also be a source of STH eggs in the soil. The STH eggs were confirmed in 43.55% of dog droppings. The most interesting finding was that the eggs of the genus Ascaris were up to 7.93% of dog droppings from localities with a low hygienic standard were positive. This study revealed that climatic factors and the WASH conditions influenced the distribution of STHs to variable degrees. In addition, ethnicity and sanitation were crucial factors in the distribution of STH infection in eastern Slovakia.

Effect of Wastewater Treatment on Bacterial Community, Antibiotic-Resistant Bacteria and Endoparasites.

Wastewater and wastewater treatment plants serve as urban reservoirs of pathogenic microorganisms. Wastewaters frequently contain bacteria, antibiotic-resistant bacteria, and developmental stages of parasites with significant zoonotic potential. Five wastewater treatment plants in the central part of Slovakia were investigated to determine the effect of treatment on bacterial community, antibiotic-resistant bacteria, and the occurrence of helminth eggs. Although all monitored chemical factors (chemical oxygen demand, biochemical oxygen demand, N-NH4, total nitrogen, and total phosphorus) in the effluent were in line with the legislative standards for discharge into public waterways, the results of minimal inhibitory concentrations show that reclaimed water harbors E. coli resistant to several commonly used antibiotics (ampicillin, piperacillin, and tazobactam, combine ampicillin and sulbactam, cefotaxime, tetracycline). The presence of endoparasite developmental stages in wastewater and sludge (Ascaris spp., Hymenolepis nana, eggs from the Ancylostomatidae family, Giardia duodenalis) indicates potential health risks for humans and workers at these sites. Treatment such as composting before applying sludge to land is necessary to reduce human pathogens.

Therapeutic and prophylactic effect of flavonoids in post-COVID-19 therapy.

The high incidence of post-covid symptoms in humans confirms the need for effective treatment. Due to long-term complications across several disciplines, special treatment programs emerge for affected patients, emphasizing multidisciplinary care. For these reasons, we decided to look at current knowledge about possible long-term complications of COVID-19 disease and then present the effect of flavonoids, which could help alleviate or eliminate complications in humans after overcoming the COVID-19 infection. Based on articles published from 2003 to 2021, we summarize the flavonoids-based molecular mechanisms associated with the post-COVID-19 syndrome and simultaneously provide a complex view regarding their prophylactic and therapeutic potential. Review clearly sorts out the outcome of post-COVID-19 syndrome according particular body systems. The conclusion is that flavonoids play an important role in prevention of many diseases. We suggest that flavonoids as critical nutritional supplements, are suitable for the alleviation and shortening of the period associated with the post-COVID-19 syndrome. The most promising flavonoid with noteworthy therapeutic and prophylactic effect appears to be quercetin.

Neglected Diseases-Parasitic Infections among Slovakian Children from Different Populations and Genotypes of Giardia duodenalis.

Children are most prone to parasitic infections. The objectives of the study were to examine the occurrence of parasitic infections in children from different populations and to perform molecular characterization of human Giardia duodenalis isolates. We examined 631 stool samples from Roma and non-Roma children for the presence of parasitic developmental stages. Samples were collected from three eastern Slovakia districts. The ages of the children ranged from 1 months to 17 years. Subsequently, the molecular characterization of human G. duodenalis isolates by PCR detected triosephosphate isomerase (tpi) and beta-giardin (bg) genes was performed. The overall prevalence of parasitic infection was 19.8%. Ascaris lumbricoides eggs were the most frequent, with an occurrence of about 13.8%. G. duodenalis cysts were present in 6.3% of samples. G. duodenalis isolates obtained from 13 children were subjected to DNA sequencing with tpi and bg genes. Five isolates were categorized as bearing subassemblage BIII, the three isolates as subassemblage BIV, one person was infected with a mixture of subassemblages BIII and BIV, four children had subassemblage AII, and one isolate revealed a structure corresponding with subassemblage AI. Our work is proof that poverty and poor hygiene contribute the most to public health problems associated with neglected parasitic diseases.

The occurence of endoparasites in Slovakian household dogs and cats.

Pets play a pivotal role as definitive or reservoir hosts for many zoonotic parasites. Dogs and cats without any clinical signs may be a carrier for the infection. In a one-year study, collected fecal samples of 257 dogs and 50 cats were examined coproscopically for endoparasite infections. Out of 307 investigated fecal samples, 107 (34.9%) were positive for the presence of the propagative stages of endoparasites In 257 dogs fecal samples, following 12 different species of endoparasites were detected: Giardia duodenalis, Cystoisospora spp., Sarcocystis spp., Hammondia/Neospora-like eggs, Angiostrongylus vasorum larvae, Capillaria aerophila, Crenosoma vulpis, Toxocara spp., Toxascaris leonina, Trichuris vulpis, Strongyloides stercoralis, and eggs from the family Ancylostomatidae. Only 4 different parasitic species were found in 50 domestic cat fecal samples - G. duodenalis cysts Cystoisospora spp., T. cati, and larvae of Aelurostrongylus abstrusus. It was confirmed that significant differences were found concerning age for G. duodenalis, T. canis, S. stercoralis, and family Ancylostomatidae. Close and frequent contact between pets and people increases the risks for the transmission of zoonotic diseases.

First report on parasites of European beavers in the Slovak Republic.

European beaver (Castor fiber L. 1758) is the biggest rodent living in Europe. It is a semi-aquatic animal known for building dams and burrows. European beaver is a potential host for a wide range of parasites and other infectious diseases. In Slovakia, there is an increasing number of beavers but the data about their parasitic fauna are missing. Our work is the first documentation about the beaver's parasitofauna in Slovakia. In a 1-year study, we collected and examined 19 beaver fecal samples from the vicinity of beaver burrows inhabiting three particular localities at the Danube, Topla, and Laborec rivers in Slovakia. In these fecal samples, 4 different species of intestinal endoparasites were detected as follows: oocysts of Cryptosporidium, cysts of Giardia, eggs of Stichorchis subtriquetrus, and eggs and larvae of Travassosius rufus. Parasites were confirmed only in samples collected at river Topla. Based on our results, we can conclude that European beaver can be an important source of parasitic contamination of surface waters especially in the localities shared by people.

First report on Giardia duodenalis assemblage F in Slovakian children living in poor environmental conditions.

BACKGROUND: Giardiasis is one of the most common gastrointestinal infections of humans and animals attributable to complex of eight morphologically identical genetic assemblages, further divided into sub-assemblages. Disease is common for a wide range of hosts and genetic characterization is needed for better understanding of multifaceted epidemiology for this protozoan parasite. The aim of this study was to identify genetic heterogeneity in assemblages and sub-assemblages of Giardiaduodenalis circulating among the children population living in deprived socioeconomic conditions. METHODS: A total of 333 stool samples from children in eastern Slovakia were collected during the period of 2015-2016 and analysed by molecular methods. Molecular characterization of G. duodenalis was performed by sequence analysis of triose phosphate isomerase gene (tpi) and glutamate dehydrogenase gene (gdh). RESULTS: G. duodenalis DNA was detected in 21 samples (6.3%), out of which 14 isolates (66.7%) belonged to assemblage B, 4 isolates (19.0%) to sub-assemblage AII and 3 isolates (14.3%) corresponded to assemblage F. As regards the determination of sub-assemblages of assemblage B, 4 isolates were characterized as sub-assemblage BIII and 6 isolates as sub-assemblage BIV. CONCLUSION: This study is the first finding of cat specific assemblage F in man not only in Slovakia, but also in Europe. The absence of molecular data about G. duodenalis in companion animals in Slovakia establishes a strong need for further investigation for potential sources of giardiasis and understanding the epidemiology will help to improve the preventive strategies in eradication of infection in this population.

Occurrence of the most common helminth infections among children in the Eastern Slovak Republic.

OBJECTIVES: Ascariasis, trichuriasis and hymenolepiasis occur primarily within poor communities with low hygiene standards. This study examined the occurrence of intestinal helminth infections among children living in two counties (Kosice and Presov) in the Eastern Slovak Republic. STUDY DESIGN: Four hundred and twenty-six children were divided into groups according to ethnicity (non-Roma and Roma), age, sex, urban/rural residency and county of residence. METHODS: Stool samples collected from participants were processed by formalin-ethyl acetate sedimentation and examined microscopically. RESULTS: The overall prevalence of infection was 16.90% and the most prevalent species was Ascaris lumbricoides (14.32%). This was followed by Trichuris trichiura (3.76%), Hymenolepis nana (0.94%) and Hymenolepis diminuta (0.23%). The odds ratio for infection was 52 times higher among Roma children compared with non-Roma children. Among Roma children, the lowest prevalence of infection was reported in adolescents aged ≥16 years. No significant differences in the prevalence of helminths were found between different sexes, or between hospitalized and non-hospitalized participants. Roma children living in urban areas had a 3.36 higher probability of infection than those living in rural areas. Among Roma children, helminth ova were found in 31.76% of the specimens from Kosice County and 19.69% of the specimens from Presov County. Among non-Roma children, there was only one positive finding in Presov County, and no cases in Kosice County. CONCLUSIONS: Important risk factors associated with helminth infections are ethnicity, county of residence and urban/rural residency. Ascariasis, trichuriasis and hymenolepiasis still occur in children with low hygiene standards, and this needs to be addressed by local authorities.

Complement regulatory protein Crry deficiency contributes to the antigen specific recall response in experimental autoimmune myasthenia gravis.

BACKGROUND: Myasthenia gravis (MG) and animal model of experimental autoimmune myasthenia gravis (EAMG) is the most common autoimmune disorder of neuromuscular transmission. The disease is caused by the breakdown of the acetylcholine receptor (AChR) which is largely due to complement activation at the neuromuscular junction (NMJ). Limited knowledge exists to the extent that complement receptor 1-related gene/protein y deficiency (Crry -/-) modulates the adaptive immune response and EAMG outcome. METHODS: Mouse EAMG was induced by s.c. administrations of purified acetylcholine receptor (AChR) to Crry -/- and age- matched WT (C57BL/6) mice. Disease severity was assessed by clinical score assessment and muscle grip strength measurements. Serum complement activity was determined by hemolytic assay. ELISA was used to detect the level of AChR specific antibodies. Splenic cells were analyzed for T and B cells subsets distribution, release of cytokines and AChR specific recall responses. Deposition of complement components at the NMJ was assessed by immunofluorescence staining. RESULTS: In comparison to WT EAMG, Crry -/- EAMG mice showed signs of augmented muscle weakness but differences, except for one time point, were not statistically significant. Serum complement activity was reduced in Crry -/- EAMG mice and no substantial changes in deposition of C3, C3b/iC3b and C5b-9 (MAC) at the NMJ between WT EAMG and Crry -/- EAMG mice were detected. Lack of Crry affected adaptive immune response. Crry -/- EAMG mice showed increases in the number of AChR specific splenic T-cells secreting IFN-γ and IL-4. Production of complement fixing antibodies (IgG2b, IgG2c) was also augmented. More Th1, Th2 and Th17 cytokines were released into the bloodstream of Crry -/- EAMG mice. CONCLUSIONS: Data suggest that Crry deficiency modulates the adaptive immune response in EAMG, but its effect on disease outcome is limited. This was due to the generally lower serum complement level caused by increased C3 turnover. Modulation of complement activity with soluble or membrane bound regulators of complement activity represents a potentially effective approach to modify autoimmune processes in MG and EAMG.

DAF/CD55 and Protectin/CD59 modulate adaptive immunity and disease outcome in experimental autoimmune myasthenia gravis.

The role of regulators of complement activity (RCA) involving CD55 and CD59 in the pathogenesis of experimental autoimmune myasthenia gravis (EAMG) remains unclear. CD55 and CD59 restrict complement activation by inhibiting C3/C5 convertases' activities and membrane attack complex formation, respectively. Actively immunized EAMG mice deficient in either CD55 or CD59 showed significant differences in adaptive immune responses and worsened disease outcome associated with increased levels of serum cytokines, modified production of acetylcholine receptor antibodies, and more complement deposition at the neuromuscular junction. We conclude that modulation of complement activity by RCA represents an alternative in controlling of autoimmune processes in EAMG.

Novel complement inhibitor limits severity of experimentally myasthenia gravis.

OBJECTIVE: Complement mediated injury of the neuromuscular junction is considered a primary disease mechanism in human myasthenia gravis and animal models of experimentally acquired myasthenia gravis (EAMG). We utilized active and passive models of EAMG to investigate the efficacy of a novel C5 complement inhibitor rEV576, recombinantly produced protein derived from tick saliva, in moderating disease severity. METHODS: Standardized disease severity assessment, serum complement hemolytic activity, serum cytotoxicity, acetylcholine receptor (AChR) antibody concentration, IgG subclassification, and C9 deposition at the neuromuscular junction were used to assess the effect of complement inhibition on EAMG induced by administration of AChR antibody or immunization with purified AChR. RESULTS: Administration of rEV576 in passive transfer EAMG limited disease severity as evidenced by 100% survival rate and a low disease severity score. In active EAMG, rats with severe and mild EAMG were protected from worsening of disease and had limited weight loss. Serum complement activity (CH(50)) in severe and mild EAMG was reduced to undetectable levels during treatment, and C9 deposition at the neuromuscular junction was reduced. Treatment with rEV576 resulted in reduction of toxicity of serum from severe and mild EAMG rats. Levels of total AChR IgG, and IgG(2a) antibodies were similar, but unexpectedly, the concentration of complement fixing IgG(1) antibodies was lower in a group of rEV576-treated animals, suggesting an effect of rEV576 on cellular immunity. INTERPRETATION: Inhibition of complement significantly reduced weakness in two models of EAMG. C5 inhibition could prove to be of significant therapeutic value in human myasthenia gravis.

The role of Rac2 in regulating neutrophil production in the bone marrow and circulating neutrophil counts.

Circulating neutrophils are persistently higher in mice deficient in the small GTPase Rac2 than in wild-type (WT) mice. Therefore, we examined the mechanisms through which the small GTPase Rac2 regulates neutrophil production and release. Lethally irradiated WT mice reconstituted with a 50:50 mixture of WT and Rac2(-/-) fetal liver cells were protected from neutrophilia, suggesting that neutrophilia is primarily because of extrinsic defects that can be corrected by WT leukocytes. However, the differential counts and numbers of leukocyte subtypes differed between Rac2(-/-) and WT cells, suggesting that Rac2 modulates leukocyte lineage distribution. Kinetic studies suggest Rac2 modulates the release of neutrophils into the circulation and does not prolong their circulating half life. The percentage of bone marrow cells that expressed the neutrophil marker Gr-1 in lethally irradiated WT or Rac2(-/-) recipients of Rac2(-/-) stem cells was greater than in recipients of WT stem cells; however, circulating neutrophil counts were higher only in Rac2(-/-) recipients of Rac2(-/-) stem cells. Rac2 mRNA was expressed in the bone marrow of WT recipients of Rac2(-/-) stem cells and in human mesenchymal stem cells. The data presented here suggest that Rac2 in hematopoietic cells regulates leukocyte lineage distribution and Rac2 in nonhematopoietic cells might contribute to regulating circulating neutrophil counts.

Extraocular muscle susceptibility to myasthenia gravis: unique immunological environment?

Extraocular muscle (EOM) is susceptible to neuromuscular junction disorders, in particular, myasthenia gravis (MG). While EOM physiological characteristics and the ocular motor system requirements contribute to the propensity of ocular motor deficits observed among patients with MG, the authors propose that EOM have immunological features that place the muscles at risk for immune attack. Genomic profiling studies have demonstrated that genes associated with the immune response are differentially expressed in EOM, with particular differences in both classical and alternative complement-mediated immune response pathways. Intrinsic complement regulators are expressed at lower levels at rodent EOM neuromuscular junctions, which would put them at risk for the complement-mediated injury that occurs in MG. In fact, systemic C inhibition in experimental autoimmune MG (EAMG) induced by administration of acetylcholine receptor (AChR) antibodies or immunization with AChR will eliminate complement deposition at junctions of other skeletal muscle, but not EOM. Also, EOM junctions have greater injury in active and passive EAMG by several measures, suggesting that the lack of complement inhibition puts the EOM at risk. Among ocular myasthenia patients, serum AChR antibody levels are low, which would support the concept that EOM junctions are more susceptible to antibody injury than are other junctions. These observations suggest that complement inhibitory therapies may prove to be particularly effective in treatment of ocular myasthenia.

Effect of complement and its regulation on myasthenia gravis pathogenesis.

Myasthenia gravis (MG) is primarily caused by antibodies directed towards the skeletal muscle acetylcholine receptor, leading to muscle weakness. Although these antibodies may induce compromise of neuromuscular transmission by blocking acetylcholine receptor function or antigenic modulation, the predominant mechanism of injury to the neuromuscular junction is complement-mediated lysis of the postsynaptic membrane. The vast majority of data to support the role of complement derives from experimentally acquired MG (EAMG). In this article, we review studies that demonstrate the central role of complement in EAMG and MG pathogenesis along with the emerging role of complement in T- and B-cell function, as well as the potential for complement inhibitor-based therapy to treat human MG.

Acute Pseudomonas challenge in cystic fibrosis mice causes prolonged nuclear factor-kappa B activation, cytokine secretion, and persistent lung inflammation.

BACKGROUND: Cystic fibrosis (CF) is characterized by an excessive and prolonged inflammatory response to Pseudomonas aeruginosa in the lung. There are high levels of cytokines and chemokines and an exaggerated PMN influx causing significant morbidity and mortality. OBJECTIVE: To compare the kinetics of the inflammatory response with the kinetics of clearance of acute bacterial challenge in the lungs of CF and wild-type (WT) mice. METHODS: We challenged CF knockout (KO) and WT mice intratracheally with P aeruginosa in suspension and evaluated bacteria counts, nuclear factor-kappaB (NF-kappaB), and inhibitor of NF-kappaB alpha protein (I-kappaBalpha) in lung tissue, cytokines, and PMN in bronchoalveolar lavage (BAL). RESULTS: Both groups of mice cleared the infection with the same kinetics. CF-KO mice had more PMN in BAL than WT mice. CF-KO mice had high concentrations of proinflammatory cytokines in BAL on days 2 and 4, whereas cytokines in BAL from WT mice were only slightly elevated. CF-KO mice failed to regenerate I-kappaBalpha once it was degraded, and consequently had prolonged and excessive activation of NF-kappaB for the entire 6-day duration of the study. In contrast, WT mice showed only slight NF-kappaB activation, which plateaued at day 4. CONCLUSION: These data suggest that NF-kappaB is dysregulated in CF lung infection and could be a good target for therapy. Prolonged responses to initial acute infections may contribute to the eventual establishment of chronic persistent inflammation. CLINICAL IMPLICATIONS: Dysregulation of the I-kappaB/NF-kappaB pathway in cystic fibrosis leads to prolonged cytokine secretion and persistent inflammation in response to acute challenges and may be important in the development of chronic lung inflammation and infection.

Role of IL-10 deficiency in excessive nuclear factor-kappaB activation and lung inflammation in cystic fibrosis transmembrane conductance regulator knockout mice.

BACKGROUND: Patients with cystic fibrosis (CF) and CF transmembrane conductance regulator knockout (CF-KO) mice are deficient in pulmonary IL-10 and have excessive inflammatory response to Pseudomonas aeruginosa infection. OBJECTIVE: We hypothesized that local IL-10 deficiency in the lung was responsible for prolonged and excessive inflammatory responses and observations of inflammation in the absence of infection. METHODS: To determine whether IL-10 deficiency could account for persistent inflammation in CF mice independent of interactions of bacteria with epithelial cells, we challenged IL-10-knockout (IL-10-KO), CF-KO, and wild-type (WT) mice intratracheally with LPS and determined the effects of IL-10 replacement in CF-KO mice. RESULTS: In response to LPS, IL-10-KO and CF-KO mice had more neutrophils and proinflammatory cytokines in bronchoalveolar lavage than WT mice. Both types of knockout mice had more profound and prolonged consumption of I-kappaB and increased activation of nuclear factor kappaB (NF-kappaB). Activated NF-kappaB persisted for 6 to 8 hours in CF-KO and IL-10-KO mice but was not detected beyond 2 hours in WT mice. IL-10 treatment of CF-KO mice attenuated the reduction in I-kappaBalpha and activation of NF-kappaB and reduced the excessive inflammation. CONCLUSION: Similarities in the responses of CF-KO and IL-10-KO mice and correction of excessive responses in CF mice by exogenous IL-10 suggest that deficiency of IL-10 may be responsible for prolonged and excessive inflammatory responses in CF. Because LPS was used as the stimulus, these excessive responses are independent of any possible differences in the interactions of bacteria with CF epithelial cells.

Functional IL-10 deficiency in the lung of cystic fibrosis (cftr(-/-)) and IL-10 knockout mice causes increased expression and function of B7 costimulatory molecules on alveolar macrophages.

Alveolar macrophages are poor APCs that only minimally express B7 costimulatory molecules. Because our previous data suggest that bronchial epithelial cells constitutively secrete IL-10, and IL-10 inhibits B7 expression in vitro, we hypothesized that this IL-10 is responsible for suppressing B7 expression on macrophages that enter the airways. Furthermore, because we have shown that cystic fibrosis (CF) lungs are deficient in IL-10, we hypothesized that bronchoalveolar macrophages (BALMs) from cystic fibrosis transmembrane conductance regulator (CFTR)(-/-) as well as IL-10(-/-) mice might express increased B7. Immunofluorescence for B7 was positive on BALMs from CF patients and CFTR(-/-) and IL-10(-/-) mice, but was negative on controls. FACS showed that 63.9% of BALMs from IL-10(-/-) mice were B7-1 positive, as were 67.4% of BALMs from CFTR(-/-) mice, whereas <7% of BALMs from wild-type controls were positive. Using BALMs to costimulate splenic T cells with anti-CD3 as a mitogen showed 9202 +/- 2107 cpm [(3)H]thymidine incorporation for BALMs from IL-10(-/-) mice and 4082 +/- 1036 cpm for BALMs from CFTR(-/-) mice, but <200 cpm with BALMs from either type of +/+ mouse. Treatment of CFTR(-/-) mice with recombinant mouse IL-10 reduced the B7 expression and costimulatory activity of the BALMs. These data suggest that the IL-10 secreted in the healthy lung may be responsible for the absence of B7 and poor costimulatory activity of BALMs and that reductions of pulmonary IL-10 in CF may enhance B7 expression and local immune responses.