Characterization of Fructose-1,6-Bisphosphate Aldolase 1 of Echinococcus multilocularis

Glycolysis is one of the important ways by which Echinococcus multilocularis acquires energy. Fructose-1, 6-bisphosphate aldolase (FBA) plays an important role in this process, but it is not fully characterized in E. multilocularis yet. The results of genome-wide analysis showed that the Echinococcus species contained four fba genes (FBA1-4), all of which had the domain of FBA I and multiple conserved active sites. EmFBA1 was mainly located in the germinal layer and the posterior of the protoscolex. The enzyme activity of EmFBA1 was 67.42 U/mg with Km and Vmax of 1.75 mM and 0.5 mmol/min, respectively. EmFBA1 was only susceptible to Fe3+ but not to the other four ions (Na+, Ca2+, K+, Mg2+), and its enzyme activity was remarkably lost in the presence of 0.5 mM Fe3+. The current study reveals the biochemical characters of EmFBA1 and is informative for further investigation of its role in the glycolysis in E. multilocularis.

Synergism therapeutic and immunoregulatory effects of Albendazole + rAd-mIL-28B against Echinococcosis in experiment-infected mice with protoscoleces

The metacestode stage of Echinococcus granulosus can cause cystic echinococcosis (CE), which still widely occurs around the world. Since the early 1970s, benzimidazoles have been shown to inhibit the growth of cysts and used to treat CE. However, benzimidazoles are still ineffective in 20%-40% of cases. In order to explore the new agents against CE, we have investigated the therapeutic effect of the recombinant adenoviral vector expressing mouse IL-28B (rAd-mIL-28B) on protoscoleces-infected mice. In our study, we successfully established the model mice which infected with protoscoleces intraperitoneally. At 18 weeks post-infection, the mice received rAd-mIL-28B (1×107 PFU) weekly by intramuscular injection for 6 weeks. Compared with the untreated control (13.1 ± 2.2 g), there was a significant reduction in cysts wet weight in rAd-mIL-28B group (8.3 ± 3.5 g) (P < 0.05), especially in Albendazole (ABZ) + rAd-mIL-28B group (5.8 ± 1.4 g) (P < 0.01). We also observed the severe damage of the germinal layer and the laminated layer of cysts after treatment. rAd-mIL-28B group showed a prominent increase in the level of Th1 type cytokines (such as IFN-γ, IL-2 and TNF-α). Meanwhile, the frequency of Foxp3+ T cells was decreased in the rAd-mIL-28B group (4.83 ± 0.81%) and ABZ + rAd-mIL-28B group (4.60 ± 0.51%), comparing with the untreated group (8.13 ± 2.60%) (P < 0.05). In addition, compared with the untreated control (122.14 ± 81.09 pg/ml), the level of IFN-γ significantly increased in peritoneal fluid in the rAd-mIL-28B group (628.87 ± 467.16 pg/ml) (P < 0.05) and ABZ + rAd-mIL-28B group (999.76 ± 587.60 pg/ml) (P < 0.001). Taken together, it suggested that ABZ + IL-28B may be a potential therapeutic agent against CE.

Defensive Impact of Co Q10 in Japanese Quail Males Raised Under Oxidative Stress Conditions

This study was investigated the protective role of (CoQ10) on reproductive dysfunction of males quail induced by hydrogen peroxide (H2O2). Forty Japanese quail male 45 days old were randomly allotted into four groups with ten replicate one per each , and treated for 28 days as follows : (G1): the first control group (G2): reserved standard ratio and (1%) H2O2 in drinking water. (G3): reserved production ratio + (100 mg Co Q10/kg diet) and (1%) H2O2 in drinking water. (G4): reserved production ratio + (100 mg Co Q10/kg diet) and normal water. The results showed that exposure of birds to H2O2 caused decrease in serum glutathione level (GSH), testosterone, luteinizing hormones (LH) and follicle stimulating hormone (FSH) hormones and this reflects on histology of testis by reducing seminiferous tubules diameter , area of germinal layer and germinal layer thickness. Whereas supplement of CoQ10 caused an increase the concentrations of theses, hormones in-group G4 as compared with group G2. It can noted that an addition was able to restore the oxidative stress birds (G3) to a state close to the natural state (G1). Though testicular histological modifications were made strides in grown-up Japanese quail male treated with CoQ10. In conclusion, the comes about of the display think about appeared that utilize of Co Q10 can easing the pernicious impacts on male regenerative work takingafter introduction toH2O, maybe through enhancement the antioxidant parameters or testicular capacities or other related endocrine organs.

The protective effect of crocin on cisplatin-induced testicular impairment in rats

Background Side effects of cisplatin (CIS) such as testicular toxicity restrict its clinical use. Instead, evidence indicates that crocin (CR) has synergistic anti-cancer potential with CIS and exhibited beneficial effects on CIS-induced hepatorenal damage. The aim of this study was to investigate the protective potential of CR against CIS-induced testicular toxicity in rats. Methods Fifty adult male Wistar rats randomly assigned to five equal groups including control, CIS, and CIS plus CR at doses of 6.25 mg/kg (CIS + CR6.25), 25 mg/kg (CIS + CR25), and 100 mg/kg (CIS + CR100). CIS and CIS + CR groups received a single intraperitoneally (i.p.) injection of CIS (7 mg/kg). CR (6.25–100 mg/kg i.p.) injections were started three days before the CIS injection and continued once a day for up to 13 days. On the 14th day, all animals were sacrificed and their blood samples and testes were removed for biochemical and histological analyses. Results Compared to the control group, CIS significantly decreased relative testis weight (0.28 vs. 0.39, p < 0.001), testosterone level (0.3 vs. 2.31 ng/mL, p < 0.001), germinal layer area (25,886 vs. 35,320 µm2, p < 0.001), superoxide dismutase (SOD) (0.9 vs.1.73 U/mg, p < 0.001) and increased testicular lipid peroxidation (3.05 vs. 15.35 nmol/mg, p < 0.001). CR at 25 mg/kg ameliorated testicular lipid peroxidation and enhanced SOD activity compared to CIS group (p < 0.05). Besides, CR treatment at the maximum dose (100 mg/kg) resulted in reversing CIS effects on testis weight, testosterone level, SOD, lipid peroxidation, and germinal layer area. Conclusions These findings demonstrated that CR co-treatment could prevent CIS-induced testicular toxicity in rats.

Dysregulation of Ovine Toll-Like Receptors 2 and 4 Expression by Hydatid Cyst-Derived Antigens

Background: Cystic echinococcosis (CE) is a zoonotic disease caused by infection with Echinococcus granulosus. Toll-like receptors (TLRs) as the first line of defense against various parasites play a critical role in sensing and triggering anti-parasite responses. Methods: The study was conducted at the Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Iran in 2019. Ovine peripheral blood mononuclear cells (PBMCs) were stimulated with hydatid cyst-derived antigens including hydatid cyst fluid (HCF), germinal layer antigens (GL), somatic and excretory/secretory (ES) products of protoscoleces (PSC). To investigate whether the expression of TLR2 and TLR4 was altered during exposure to these antigens, PBMCs were stimulated with two different concentrations at different time points. Results: After exposure of PBMCs to ES and somatic antigens of protoscoleces (PSC) the expression of TLR2 and TLR4 was down-regulated in comparison with control group. Similarly, HCF markedly down-regulated TLR2 and TLR4 transcripts independent of dose and time. GL antigens significantly down-regulated TLR2, while TLR4 expression was up-regulated as compared with control group. Conclusion: Hydatid cyst-derived antigens could dysregulate the expression of TLR2 and TLR4 in ovine PBMCs, suggesting a possible mechanism to suppress host immunity to establish chronic infection.

Cerebral Coenurosis of a Long-Tailed Goral, Naemorhedus caudatus, in Korea

We intended to describe a case of cerebral coenurosis in a long-tailed goral, Naemorhedus caudatus, from Hwacheon-gun, Gangwon-do (Province), in the Korea. The goral, a 10-year-old male, was suffering from neurological symptoms, such as turning the circle to one side without lifting the head straight, and died at 30 days after admission to the wildlife medical rescue center in Chuncheon-si, Gangwon-do. A fluid-filled cyst was detected in the left cerebral hemisphere by computed tomography and magnetic resonance imaging. The cyst removed from the deceased goral was transparent, about 3×3 cm in size, contained a clear fluid and approximately 320 protoscolices invaginating from the internal germinal layer. The protoscolex had 4 suckers and a rostellum with 28 hooklets arranged in 2 rows. By the present study, a case of cerebral coenurosis was first confirmed in a long-tailed goral, N. caudatus, from Gangwon-do, in Korea. The residents frequently exposed in the sylvatic environment should be careful the accidental infections of zoonotic metacestode of Taenia multiceps, Coenurus cerebralis, in Korea.

IMPACTS OF QUERCETIN AND MERCAPTOTHION ON MALE REPRODUCTIVE PARAMETERS OF RATS

Quercetin is an herbal polyphenol with valuable antioxidant properties. Mercaptothion is categorized as organophosphates which can generate free radicals and induce male fertility disorders. This study was aimed to assess the impacts of Quercetin against destruction of male fertility parameters induced by Mercaptothion. 64 male rats were randomly assigned into 8 groups; control, and Mercaptothion (250 mg/kg) groups; Quercetin groups (7.5, 15, and 30 mg/kg) and Mercaptothion + Quercetin (7.5, 15, and 30 mg/kg). Treatments were administered intraperitoneally (Mercaptothion) and orally (Quercetin) daily for 65 days. The sperm parameters, testis malondialdehyde (MDA), total antioxidant capacity (TAC), testosterone level and height of germinal layer were evaluated. Expressions of p53, caspase-3, Bax, and Bcl-2 were measured through real-time quantitative PCR. Values of all factors were reduced significantly except the MDA level (which increased) in Mercaptothion group compared to the control group (p<0.001). Studied criteria in groups of Quercetin and Quercetin + Mercaptothion in whole doses increased significantly except MDA level (which reduced) compared to the Mercaptothion group (p<0.001). Also, downregulated levels of p53, caspase-3, and Bax genes and unregulated levels of Bcl-2 gene expression were detected in control and the sixth treatment groups significantly in Quercetin group compared to the Mercaptothion group (p<0.001). No significant alterations were detected in Quercetin groups compared to the control group (p>0.05). Quercetin reduced toxic effects of Mercaptothion on male fertility parameters.

Comparative analysis of miRNA expressions in different developmental stages of Echinococcus granulosus in mono-phasic and di-phasic culture systems

Background:: The dog tapeworm, Echinococcus granulosus, is a zoonotic parasite affecting human and livestock across the globe. Basic research on the molecular biology and genetics of E. granulosus improves our understanding of the biology and potential drug targets in various developmental stages of E. granulosus in both definitive and intermediate hosts. There has been increasing interest in identification of microRNAs in parasitic organisms. The purpose of the current study was to compare the activity of a selected profile of miRNAs in different developmental stages of E. granulosus. Methods:: Different developmental stages of the parasite were obtained from ex vivo as well as in vitro cultured E. granu-losus. MicroRNAs were extracted from the ex vivo germinal layer and invaginated protoscoleces as well as the in vitro gen-erated microcysts, evaginated protoscoleces and strobilated worms. Expression of the selected miRNAs was evaluated by RT-qPCR for each stage. Results:: Four out of five miRNAs were present and active in different developmental stages of E. granulosus. A significant over-expression of miR-61 was observed in germinal layer and during the protoscolex transformation into the microcysts, however miR-10 was more expressed in the mature strobilated forms than the other stages. Let-7 and miR-3489 showed a high expression in germinal layer. Conclusion:: Differential expression of four miRNAs among different in vitro and ex vivo developmental stages of E. granu-losus was documented in the present study. Further experimental investigations are required to elucidate the probable role of the miRNAs in bi-directional differentiation of protoscoleces either into the strobilated worm or to a secondary hydatid cyst and the potential of these miRNAs as drug targets.

Ultrastructure of Human Fertile Hydatid Cysts Using a Scanning Electron Microscope (SEM)

Introduction: Echinococcosis and hydatidosis caused by the metacestode of Echinococcus granulosus are among the most important zoonotic diseases in the world. This study aims to study the ultrastructure of fertile hydatid cysts that infect humans using a scanning electron microscope (SEM). Materials and Methods: Twenty samples of human fertile hydatid cysts were collected from the human liver and lung after performing surgery operations and examined with an SEM. Results: The results of the electron microscopy with different magnifications revealed that the laminated layer (LL) consists of sheets that appeared more compact and aligned. The brood capsules appeared, consisting of a net of finger-shaped structures that emerged from bulges of various sizes and shapes. Conclusion: Under a transmission electron microscope, it was found that the LL had a coherent and flexible structure, settling on a three-dimensional microscopic network of hydrophilic fibers, with high humidity. These fibers were arranged irregularly and had a diameter of about 10 nm; therefore, the fibers adjacent to the germinal layer (GL) were possibly attached to microtriches of tegument, which reached a thickness of 1 mm in the LL.

Efficacy of novel albendazole salt formulations against secondary cystic echinococcosis in experimentally infected mice

In this study, we evaluated the efficacy, expressed as a mean weight decrease of the whole echinococcal cyst mass, of novel benzimidazole salt formulations in a murine Echinococcus granulosus infection model. BALB/c mice were intraperitoneally infected with protoscoleces of E. granulosus (genotype G1). At 9 months post-infection, treatment with albendazole (ABZ), ricobendazole (RBZ) salt formulations, and RBZ enantiomer salts (R)-(+)-RBZ-Na and (S)-(−)-RBZ-Na formulations were initiated. Drugs were orally applied by gavage at 10 mg kg−1 body weight per day during 30 days. Experimental treatments with benzimidazole sodium salts resulted in a significant reduction of the weight of cysts compared to conventional ABZ treatment, except for the (S)-(−)-RBZ-Na enantiomer formulation. Scanning electron microscopy and histological inspection revealed that treatments impacted not only the structural integrity of the parasite tissue in the germinal layer, but also induced alterations in the laminated layer. Overall, these results demonstrate the improved efficacy of benzimidazole salt formulations compared to conventional ABZ treatment in experimental murine cystic echinococcosis.