The influence of albendazole on the cystolic glutatione-S-transferase [GST] and on the expression of Toll-like receptors [TLR2 and TLR4] during the intestinal phase of trichinellosis

Research into changes in the activity and kinetics of the reaction catalysed by the important detoxificating enzyme, glutathione-S-transferase (GST), in the intestinal phase of trichinellosis as presented in the present study is a pioneering achievement. In every instance of infection – or if infection is only suspected – the patient receives anthelmintics that are effective against mature intestinal forms, newborn larvae and encysted larvae of Trichinella spiralis. This group of drugs includes benzimidazoles such as albendazole. The primary action of these drugs concerns β-tubuline. The functions of microtubules are damaged by bonding β-tubuline with benzimidazoles, which inhibits the polymerisation of microtubules. Newly considered fields of action of benzimidazoles on the pathological changes occurring in the host during the course of parasitic diseases are the following elements taking part in the immunological defence against a parasitic infection: for example, detoxificating enzymes such as glutathione-Stransferase (GST), and Toll-like receptors (TLRs). Occurrence of cell resistance to drugs is attributed to certain enzymes, (such as GST), which perform the detoxification of these drugs and facilitate their removal from cells through a pump dependent on compounds coupled with glutathione. In the case of the intestinal phase of trichinellosis, GST may remove toxins secreted by both the adult and newborn larvae of T. spiralis. The structure of a dimeric enzyme of the host, such as GST, may be impacted by various factors, for example excretory-secretory products (ES), as well as drugs used. Over the past few years, the interest of researchers in the innate immune response as the first line of defense of vertebrates against of pathogens, was increased. The number of works devoted to the role of Toll-like receptors, which enable the immediate commencement of protective action by the host against various infections without engaging lymphocytes, is on the noticeable. The experimental material consisted of small and large intestine from BALB/c mice infected with T. spiralis strain MSUS/PO/60/ISS3. Mice were inoculated orally with about 400 T. spiralis larvae per mouse. On the 2nd day post-infection, the mice were treated with albendazole, in a dose of 0.5 mg/mouse. The activity and kinetics of the GST were determined spectrophotometrically. An evaluation of the expression of TLRs (TLR2 and TLR4 on the mRNA level) was examined using the RT-PCR technique with the appropriate starter sequences for TLR2 and TLR4. The presence of Toll-like receptors and their expression on the protein level was confirmed using the immunohistochemical method (Santa Cruz ABC Stainig system sc-2023 test) with specific polyclonal antibodies antiTLR2 and anti-TLR4. My results pointed to the participation of GST, as well as Toll-like receptors, in defence processes of the host during the intestinal phase of trichinellosis in mice. The adult and newborn larvae of T. spiralis present in the intestines of infected mice are a source of excretory-secretory (ES) products, which appear to stimulate the activity and change catalytic properties of conformers GST. It is supposed that ES products secreted by the parasite acts on GST as an allosteric activator. Additionally, the presence of Toll-like receptors (TLR2 and TLR4) in the intestinal phase of trichinellosis was investigated. Thus, the infection with T. spiralis leads to strong biochemical reactions with the involvement of GST, and immunological reactions with the involvement of TLR2 and TLR4 receptors. Administration of albendazole, resulted in 2-fold decrease of the total activity of GST in comparison with its activity in infected and untreated animals. Also, drug administration had weakened expression of gene of TLR4 and TLR2 in the large intestine, and expression of gene of TLR2 in the small intestine. My results confirmed destructively action of albendazole on T. spiralis (resulted in 4-fold decrease in the intensity of infection measured in muscles). The destruction by the drug of a adult and newborn larvae of T. spiralis lowers the quantities of their ES products stimulated the activation of the host’s response. This is apparent as weakening the innate immune response, in which take part Toll-like receptors play a role, and in biochemical processes with the participation of the GST enzyme. In summary my results contribute new data on the participation of Toll-like receptors and GST in the defence mechanisms of the host during in the intestinal phase of experimental trichinellosis in mice

Trichinella spiralis: impact on the expression of Toll-like receptor 4 (TLR4) gene during the intestinal phase of experimental trichinellosis

Introduction: Toll-like receptors (TLRs) play a key role in the rapid activation of the innate immune response to a variety of pathogens. The aim of this study was to evaluate the effect of Trichinella spiralis infection on the level of expression of the tlr4 gene in mouse intestines during the intestinal phase of experimental trichinellosis

New Primers for Fast Detection of Giardia duodenalis Assemblages A and B Using Real-time PCR

Giardia duodenalisis one of the six Giardia species and itis the most common, cosmopolitan flagellate that infects humans and many species of animals. This species exhibits considerable genetic diversity; to date, eight assemblages (A–H) have been defined. These assemblages differ in host specificity: assemblages A and B have been found in both humans and in many animal species. Mixed infections with Giardia (A and B) assemblages have been reported in humans and in animals. Many molecular techniques are effective and rapid for the detection of G. duodenalis and also for the determination of genetic variability of isolates in clinical and environmental samples. In this context, the aim of this study was to design new assemblage-specific primers for rapid detection and identification of G. duodenalis assemblages A and B and both of these assemblages simultaneously using quantitative real-time polymerase chain reaction (qPCR). Fragments of glutamate dehydrogenase and triose phosphate isomerase were used as targets in the design of primers. In conclusion, the use of G. duodenalis assemblage-specific primers designed in this study allows quick identification of human infectious G. duodenalis assemblages A and B as well as mixed AB assemblages in a sample without further sequencing of the amplification products, which reduces the cost of study and the waiting time for the results

New Primers for Fast Detection of Giardia duodenalis Assemblages A and B Using Real-time PCR

Solarczyk, Piotr, Wojtkowiak-Giera, Agnieszka, Hołysz, Marcin, Słodkowicz-Kowalska, Anna, Jagodziński, Paweł P., Stojecki, Krzysztof, Rocka, Anna, Majewska, Anna C., Skrzypczak, Łukasz (2018): New Primers for Fast Detection of Giardia duodenalis Assemblages A and B Using Real-time PCR. Acta Protozoologica 57 (1): 43-48, DOI: 10.4467/16890027AP.18.003.8397, URL: https://www.mendeley.com/catalogue/7aba2f6e-e1ea-3142-9412-7b469479f583