LE Strain Research Articles

Innate immune receptors are differentially expressed in mice during experimental Schistosoma mansoni early infection.

The impact of Schistosoma mansoni infection over the immune response and the mechanisms involved in pathogenesis are not yet completely understood. This study aimed to evaluate the expression of innate immune receptors in three distinct mouse lineages (BALB/c, C57BL/6 and Swiss) during experimental S. mansoni infection with LE strain. The parasite burden, intestinal tissue oogram and presence of hepatic granulomas were evaluated at 7- and 12-weeks post infection (wpi). The mRNA expression for innate Toll-like receptors, Nod-like receptors, their adaptor molecules, and cytokines were determined at 2, 7 and 12 wpi in the hepatic tissue by real-time quantitative polymerase chain reaction (qPCR). Swiss mice showed 100% of survival, had lower parasite burden and intestinal eggs, while infected BALB/c and C57BL/6 presented 80% and 90% of survival, respectively, higher parasite burden and intestinal eggs. The three mouse lineages displayed distinct patterns in the expression of innate immune receptors, their adaptor molecules and cytokines, at 2 and 7 wpi. Our results suggest that the pathogenesis of S. mansoni infection is related to a dynamic early activation of innate immunity receptors and cytokines important for the control of developing worms.

Experimental infection with Schistosoma mansoni isolated from the wild rodent Holochilus sciureus shows a low parasite burden but induces high schistosomiasis severity in BALB/c mice.

Wild mammals, especially rodents, can participate in the life cycle of Schistosoma mansoni; however, the impact of these parasite strains on the severity of schistosomiasis remains unclear. The aim of this study was to comparatively evaluate the parasitological and immunopathological alterations induced by an S. mansoni strain isolated from the wild rodent Holochilus sciureus (HS strain) and a parasite strain isolated from a human (LE strain) in experimentally infected mice. Male BALB/c mice were subcutaneously infected with 50 cercariae/mouse of either the HS or the LE strain and were evaluated for 12 weeks. In the experimental groups, the parasite burden was estimated by worm and egg (feces and tissues) count, and immunopathological alterations were evaluated in the liver and intestines. Compared to experimental infection with the LE parasite strain, HS-infected mice showed reduced number of parasite worms but higher fecundity rate, significant reduction in IL-5, IL-10 and IL-13 concentrations, lower EPO-activity in liver homogenate and higher concentrations of TNF-α, IFN-γ, IL-12 and IL-17 in the small intestine homogenate. Moreover, HS infection resulted in higher concentrations of NO end-products in both the liver and intestine, suggesting a predominance of the Th1/Th17 immune response. HS-infected mice also showed higher plasma transaminase levels, formed larger granulomas, and had a higher mortality rate in comparison with LE-infected mice. Data indicate that BALB/c mice infected with the HS strain of S. mansoni showed reduced susceptibility to the parasite but stronger tissue inflammation and high disease severity.

Insights from the shotgun-based proteomics analysis of Biomphalaria glabrata.

Biomphalaria glabrata is an important host in the transmission of human schistosomiasis in the Caribbean and South America. Therefore, it is of interest to analyse the proteome data of Biomphalaria glabrata hemolymph to identify immunity related proteins in host-pathogen relationship. We used shotgun proteomic and bioinformatic analyses of the non-depleted and depleted [0.5 and 0.75% Trifluoroacetic acid (TFA) depletion] hemolymph of B. glabrata (LE strain). Analysis showed 148 proteins from the hemolymph. 148 were obtained from the 0.5% TFA-depleted sample. 62 proteins follow this from the 0.75% TFA-depleted sample. However, only 59 were found from non-depleted hemolymph. A number of proteins were identified from the hemolymph of this schistosomiasis snail vector linked to immunity related functions. This provides insights to the understanding of schistosome-snail interaction.

Antiparasitary Activity Of The Juglone Compound: A Narrative Review

Objective: To report, based on the literature, the action of the compound 5-hydroxy-1,4-naphthoquinone against parasites (protozoa and helminths) that affect humans. Methods: This is a narrative review that used Pubmed and Google Scholar as a data tool. This work included articles published until September 2020 that were directly related to the use of the compound juglone in antiparasitic trials. Results: The compound juglone demonstrated promising effects as a human and animal antiparasitic substance. In protozoa, the Apicomplexo Toxoplasma gondii parasite showed a high mortality rate in concentrations of juglone in the nanomolar range. The juglone showed an average inhibitory concentration (IC50) of 1.62 µM, >100 µM, and 2.02 µM µM for Trypanosoma cruzi, T. brucei rhodesiense, and Leishmania donovani, respectively. Also, the juglone showed antihelmintic activity on Hymenolepis nana in mice, and on adult worms of Schistosoma mansoni (LE strain) with IC50 34.16 µM, 32.14 µM, and 25 µM in the 24h, 48h, and 72 h, respectively. Conclusion: The results published so far show the in vitro antiparasitic potential of juglone, and the need for further studies on the specific mode of action that interacts with parasites. Besides, the literature is still limited to studies that evaluate in vivo the compound juglone, requiring better information on its interaction with living organisms.

Phenotypic plasticity of maleSchistosoma mansonifrom the peritoneal cavity and hepatic portal system of laboratory mice and hamsters

Morphometric analysis of Schistosoma mansoni male worms obtained from AKR/J and Swiss mice was carried out. Rodents infected by the intraperitoneal route with 80 cercariae of the schistosome (LE strain) were killed by cervical dislocation at 45 and 60 days post-infection and both peritoneal lavage and perfusion of the portal system were performed for the recovery of adult worms. Characteristics including total body length, the distance between oral and ventral suckers, extension of testicular mass and the number of testes were considered in the morphological analysis. Changes that occurred in S. mansoni recovered from the peritoneal cavity or from the portal system of AKR/J and Swiss mice included total body length and reproductive characteristics. Significant morphometric alterations were also observed when worms recovered from the portal system of both strains of mice were compared with the schistosomes obtained from hamsters (Mesocricetus auratus), the vertebrate host in which the LE strain had been adapted and maintained by successive passages for more than four decades. The present results reinforce the idea that S. mansoni has high plastic potential and adaptive capacity.

Hybridism between Biomphalaria cousini and Biomphalaria amazonica and its susceptibility to Schistosoma mansoni

Molecular techniques can aid in the classification of Biomphalaria species because morphological differentiation between these species is difficult. Previous studies using phylogeny, morphological and molecular taxonomy showed that some populations studied were Biomphalaria cousini instead of Biomphalaria amazonica. Three different molecular profiles were observed that enabled the separation of B. amazonica from B. cousini. The third profile showed an association between the two and suggested the possibility of hybrids between them. Therefore, the aim of this work was to investigate the hybridism between B. cousini and B. amazonica and to verify if the hybrids are susceptible to Schistosoma mansoni. Crosses using the albinism factor as a genetic marker were performed, with pigmented B. cousini and albino B. amazonica snails identified by polymerase chain reaction-restriction fragment length polymorphism. This procedure was conducted using B. cousini and B. amazonica of the type locality accordingly to Paraense, 1966. In addition, susceptibility studies were performed using snails obtained from the crosses (hybrids) and three S. mansoni strains (LE, SJ, AL). The crosses between B. amazonica and B. cousini confirmed the occurrence of hybrids. Moreover, hybrids can be considered potential hosts of S. mansoni because they are susceptible to LE, SJ and AL strains (4.4%, 5.6% and 2.2%, respectively). These results indicate that there is a risk of introducing schistosomiasis mansoni into new areas.

Schistosoma mansoni: a method for inducing resistance to praziquantel using infected Biomphalaria glabrata snails

To elucidate the mechanisms of antischistosoma resistance, drug-resistant Schistosoma mansoni laboratory isolates are essential. We developed a new method for inducing resistance to praziquantel (PZQ) using successive drug treatments of Biomphalaria glabrata snails infected with S. mansoni. Infected B. glabrata were treated three times with 100 mg/kg PZQ for five consecutive days with a one-week interval between them. After the treatment, the cercariae (LE-PZQ) produced from these snails and the LE strains (susceptible) were used to infect mice. Forty-five days after infection, mice were treated with 200, 400 or 800 mg/kg PZQ. Thirty days post-treatment, we observed that the mean number of worms recovered by perfusion was significantly higher in the group of mice infected with the LE-PZQ isolate treated with 200 and 400 mg/kg in comparison to the LE strain with the same treatment. Moreover, there was a significant difference between the ED50 (effective dose required to kill 50% of the worms) of the LE-PZQ isolate (362 mg/kg) and the LE strain (68 mg/kg). In the in vitro assays, the worms of the LE-PZQ isolate were also less susceptible to PZQ. Thus, the use of infected snails as an experimental model for development of resistance to S. mansoni is effective, fast, simple and cheap.

Portevin – Le Chatelier effect and strain behavior of austenitic steel 02Kh18N8G2FV2AB at high temperatures

The strain behavior of austenitic steel 02Kh18N8G2FV2AB in static tensile tests is studied in a wide range of temperatures (23 - 740°C) and deformation rates (1.3 × 10 -5 - 1.3 × 10 -2 sec -1 ). The influence of the Portevin - Le Chatelier effect on the strength and ductility of steel 02Kh18N8G2FV2AB at elevated temperatures is determined. The relation between the characteristics of static strength and the creep resistance of the steel is considered.

The five choice serial reaction time task: Comparison between Sprague–Dawley and Long–Evans rats on acquisition of task, and sensitivity to phencyclidine

The 5-choice serial reaction time task (5-CSRTT) allows examination of multiple aspects of cognition/executive functions (attention/impulsivity/ perseveration). Most 5-CSRTT studies are performed with pigmented (i.e. Long–Evans: LE) rats; however, albino strains (i.e. Sprague–Dawley: SD) are more commonly used in behavioural pharmacology experiments. Hence, we compared 5-CSRTT performances of SD and LE rats and their sensitivity to acute phencyclidine (PCP, 1–2.5 mg/kg). SD required significantly fewer sessions (35 versus 50) than LE rats for task acquisition, especially at shortest stimulus light duration (1 s). However, once trained, under vehicle conditions, both strains performed similarly. In contrast, PCP treatment differentially affected the two strains. Thus, whilst percentage of accuracy was decreased for both strains, in SD rats number of premature responses was more markedly decreased, whereas omissions and latency time to correct responses were more notably increased. In addition, PCP monotonically diminished in SD, but augmented (1–1.5 mg/kg) in LE rats compulsive responding. To summarize, under our experimental conditions, the SD offer advantages over LE strain for speed of acquisition of 5-CSRTT. Once trained, basal performances of both strains were equivalent and stable enough for challenge with pharmacological compounds. However, PCP differentially affected the strains on several parameters considered.

Morbidity due to Schistosoma mansoni - Entamoeba histolytica coinfection in hamsters (Mesocricetus auratus)

Data on Schistosoma mansoni-Entamoeba histolytica coinfection are scarce in the literature. In the present study, hamsters that had been infected for 70 days with Schistosoma mansoni (LE strain) were inoculated via the portal vein with two strains of trophozoites of Entamoeba histolytica: ICB-EGG (highly virulent) and ICB-RPS (non-virulent). The most evident result of coinfection was increased morbidity and mortality, in comparison with either of the infections alone. Histologically, there were no evident signs of interaction between these two infections. The morphological findings of schistosomal granuloma and amoebic abscesses in the liver were similar to those seen in the respective single-infection controls. However, there was severe wasting of the animals with both infections and greater numbers of amoebic lesions in their livers. The results obtained indicated that schistosomiasis aggravates the course of amoebiasis in hamsters.

Reappraisal of experimental infections with cercariae and schistosomula of a brazilian strain of Schistosoma mansoni in mice

The present investigation involves a reevaluation of previous results obtained after experimental infection of Swiss Webster mice with cercariae and schistosomula of the Schistosoma mansoni LE strain maintained under laboratory conditions. Three experimental groups of mice were considered: the animals of the first group were percutaneously (ring method) infected with cercariae, those of the second were subcutaneously inoculated with cercariae and the mice of the third were inoculated by the same route with schistosomula transformed in vitro. The data obtained so far indicated that the most effective method of infection is the subcutaneous injection with schistosomula, with a mean adult worm burden recovery of 54.1% when compared to the abdominal percutaneous and subcutaneous routes of infection with cercariae, in which the values were 36.7% and 32.4%, respectively. This suggests that, in experimental infections of SW mice with a LE S. mansoni strain, the skin is to be considered an effective attrition site in the percutaneous route, whereas in the case of inoculation with cercariae, a small amount of larvae fails to be transformed into viable schistosomula, possibly due to skin phase avoidance. A brief discussion about attrition sites and elimination of larval S. mansoni worms in mice is presented.

Biomphalaria tenagophila: dominant character of the resistance to Schistosoma mansoni in descendants of crossbreedings between resistant (Taim, RS) and susceptible (Joinville, SC) strains

The aim of the present work was to study parasitological, molecular, and genetic aspects in descendants of crossbreedings between a totally resistant Biomphalaria tenagophila strain (Taim, RS) and another one highly susceptible (Joinville, SC) to Schistosoma mansoni. Descendants F1 and F2 were submitted to S. mansoni infection (LE strain). The susceptibility rates for individuals from Group F1 were 0 to 0.6%, and from Group F2 was 7.2%. The susceptible individuals from Group F2 discharged a lower number of cercariae, when compared with the susceptible parental group, and in 2 out of 9 positive snails the cercarial elimination was discontinued. In order to identify genetic markers associated with resistance the genotype of parental snails and their offspring F1 and F2 were analyzed by means of the randomly amplified polymorphic DNA method. Nevertheless, it was not possible to detect any marker associated to resistance, but the results showed that in the mentioned species the resistance character is determined by two dominant genes.

Presence of a modifier gene(s) affecting early renal carcinogenesis in the Tsc2 mutant (Eker) rat model

The Eker (Tsc2 mutant) rat model of renal carcinoma is an example of Mendelian dominantly inherited predisposition to a specific cancer. Effects of genetic background on renal carcinogenesis in the Eker rat model (Eker/Eker > Eker/BN strain) indicate the presence in the BN rat genome of a modifier gene(s) that suppresses tumorigenesis. The identification of such a modifier gene(s) might help clarify the diversity of tuberous sclerosis in humans. i) We found that preneoplastic lesions in 8-week-old F1 rats [(Eker x LE) and (Eker x BN)] were more numerous in the LE strain than in the BN strain although the difference was not large. ii) We next administered N-ethyl-N-nitrosourea (ENU; single injection, i.p.) at the age of 4 weeks to amplify the strain difference in tumorigenesis, as we had done in an earlier study to identify the predisposing gene. iii) This experiment was also done in BN congenic Eker rats to confirm the strain difference in tumorigenesis. Preneoplastic lesions were fewer in BN congenic rats than in Eker rats by a factor of 100. We used this ENU system to perform a backcross experiment [F1(Eker x BN) x Eker] and finally succeeded in mapping a new modifier locus on rat chromosome 5 (the LOD score of the D5Rat12 was 3.13).

Populational structure of Schistosoma mansoni assessed by DNA microsatellites

DNA microsatellites were used as molecular markers to analyse the population structure of the laboratory LE strain and of 10 field isolates of Schistosoma mansoni, the aetiologic agent of schistosomiasis. Out of 16,000 DNA sequences analysed in databases, 622 microsatellite loci were identified in 481 sequences (3.0%). The AT repetitions were the most frequent, followed by AAT and AC. Six loci showing perfect repetitions were selected and used in the polymerase chain reaction to evaluate polymorphisms in the number of repeats. Two groups of worms were studied. The first group consisted of 78 individuals, 39 of each sex, of the LE strain. The second group of worms consisted of 10 field isolates: seven from humans and three from snails. Four of the six loci were polymorphic, containing 11–17 alleles per locus. No linkage disequilibrium was observed among loci and none of the loci was sex linked. In both groups of worms, a significant deviation from Hardy–Weinberg equilibrium was observed. The observed heterozygosity was always lower than the expected one. The polymerase chain reaction primers were S. mansoni specific. The LE strain showed a lower total number of alleles or a lower average number of alleles/polymorphic locus than the field isolates, suggesting that 41 years of laboratory maintenance exerted selective pressure on the LE strain. The S. mansoni populations from the field were most genetically undifferentiated (RST<0.027), suggesting a high gene flow among them. Our results showed the usefulness of microsatellites for population analysis of S. mansoni, offering a new alternative for a better understanding of schistosomiasis epidemiology.

Schistosoma mansoni: the action of lovastatin on the murine model

Aimed to evaluate the lovastatin action on Schistosoma mansoni oviposition, infected mice with 100 plus minus 10 cercariae of the LE strain were used. Thirty days after infection the animals were treated with 100, 200 and 400mg/kg of lovastatin, per os, during five consecutive days and then sacrificed 7, 15, 30 or 60 days after treatment. We analyzed: distribution of worms in mesenteries and liver; mortality of worms in the liver; alteration of the oogram; eggs counting in the jejunum and liver; presence of intrauterine eggs and morphology of the worms from the treated and control groups (infected and not treated animals). Significant statistical differences were found between treated and control groups when the presence of intrauterine eggs was considered and also, alteration on the oogram, eggs at different stages of development in jejunum, liver and in the body length of males and females. The morphological study of the worms showed that the degenerative modifications occurred, mainly in the reproductive system, with reduction and alteration of the viteline follicles and the ovary of the females. Also modifications in the males testicles were observed. The results suggest that the drug under study reduces, considerably, the female S. mansoni, oviposition increases the worms' size, leads to alteration in the reproductive system of males and females. It also may induce death of a significant part of the worm populations at the dose of 400mg/kg.

Resistance of Biomphalaria occidentalis from Varzea das Flores dam, Minas Gerais, to Schistosoma mansoni infection detected by low stringency polymerase chain reaction.

Biomphalaria occidentalis Paraense, 1981 from Varzea das Flores dam, MG, Brazil, was exposed to infection with Schistosoma mansoni. Individual infection was performed with 140 B. occidentalis and 100 B. glabrata snails using LE and SJ strains. Two groups of B. occidentalis were killed after seven day-miracidia exposure to detect S. mansoni DNA, through the low stringency polymerase chain reaction (LS-PCR), and were negative. The infection rates were 69.2% (LE strain) and 96.7% (SJ strain) for B. glabrata and 0% for B. occidentalis. LS-PCR enabled early resistance diagnosis.

Premixed Edge-Flames in Spatially-Varying Straining Flows

Premixed-gas flames subject to steady spatially-varying straining flows were studied to examine one aspect of premixed flames in strongly turbulent flows where strain rate gradients are present and local strain rates may be high enough to cause local flame-front extinguishment. The spatially-varying straining flows were created using a counterflow slot-jet burner with slightly non-parallel jet exits. When the flow configuration was premixed combustible gas vs. cold inert gas, so that only a single flame was produced, steady flame “edges” could be created where the flame would exist in the low-strain region but would be extinguished in the high-strain region. When the flow configuration was premixed gas vs. premixed gas, twin flames would exist in the low-strain region that converged to a corner-like tip in the high-strain region. For both configurations the local strain at the location of the stationary flame edge was somewhat lower than the strain required to extinguish flames in the same mixture subject to a spatially uniform strain. The difference was greater for the twin-flame configuration, particularly at high Lewis number (Le). Due to diffusive-thermal instabilities, cellular flames were observed at low Le and travelling-wave patterns were observed at high Le. Le effects also led to the formation of isolated “flame tubes” rather than continuous fronts at sufficiently low Le and high strain rates. All of these results are consistent with recent theoretical predictions. These results indicate that “laminar flamelet” models of premixed turbulent combustion may be reasonably accurate for single flames over a wide range of Le and twin flames with Le close to unity, even at conditions approaching those where local flame quenching occurs, but may not be accurate for twin-flames except for Le near unity. This finding is somewhat different from previous experimental and theoretical results for nonpremixed edge-flames, where more substantial differences between uniform flames and edge-flames were found for all Le.

Ｍｏｄ．９Ｃｒ‐１Ｍｏ鋼の高温多軸低サイクル疲労および裂伝ぱ挙動

This paper describes the low cycle fatigue criterion of Mod.9Cr-1Mo steel and the fracture mode under multiaxial stress/strain states. Multiaxial tension-torsion low cycle fatigue tests were carried out using plain Mod.9Cr-1Mo tubular specimens at 823K. Plain, surface notched and through notched specimens were also torsionally fatigued at 823K and room temperature to examine the crack propagation direction. LE strain and the equivalent strain based on crack opening displacement were a suitable strain parameter for correlating the multiaxial low cycle fatigue lives with a small scatter. Mises' stress was a suitable stress parameter for the correlation. Main crack of the smooth, surface notched and through notched specimens propagated in the maximum shear strain direction in torsion tests, so there was no influence of the notches on the crack propagation direction. Surface oxidation has no effect on the main crack propagation direction, but it has an influence on the subcrack propagation direction. Oxide films were brittlely broken in the principal direction.

The resistance of Biomphalaria glabrata to Schistosoma mansoni infection: variations in the prepatent period and in compatibility

Biomphalaria glabrata from Belo Horizonte, Minas Gerais, Brazil, reared in laboratory, has a level of infection of 90% when exposed to 20 miracidia of the autochibonous LE strain. The prepatent period was of 5 to 7 weeks whereas 5 to 10% of exposed snails do not shed cercariae. The eggs of negative snails were collected and the progeny was again submitted to individual infection with 20 miracidia. The mean of infection from F14 to F20 was of 43.6%. Histological sections from F12, F14 and F15 snails showed tissue reactions in those specimens shedding less than 10 cercariae. A prepatent period of 17 to 32 weeks was observed in 35 (17.9%) of 195 infected snails. The index of cercariae of control was extremely compatible and for F12, F13 and F15 snails varied from very compatible class V to compatible class III, showing less compatibility in selected snails.

Resistance to oxamniquine of a Schistosoma mansoni strain isolated from patient submitted to repeated treatments.

A strain of Schistosoma mansoni (R1) was isolated from patient previously submitted to four treatments with oxamniquine, and to another one with praziquantel. The results obtained with chemotherapeutic test, by using oxamniquine in mice infected with the strains R1 and LE (standard), showed an evident resistance to the drug in worms of the strain R1. Thus, at the dose of 250 mg/kg oxamniquine, all mice (17) infected with the LE strain did not show surviving worms, whereas 12 out of 17 mice infected with the R1 strain presented surviving worms. At the dose of 200 mg/kg, the LE strain showed recovery rates of 1.06% and 20.58%, whereas the R1 strain presented 18.57% and 61.14%, for male and female worms, respectively. At the dose of 100 mg/kg, the recovery of male worms was 2.6% for the LE strain, and 29.9% for the R1 strain. At the same dose, the recovery of females did not show statistically significant differences between the two strains (LE = 76.38%, R1 = 79.12%). Praziquantel showed similar antischistosomal activity against both studied strains, when administered at the dose of 500 mg/kg.