S100 proteins and their implication in the thalamus: an integrative review / Las proteínas S100 y su implicación en el tálamo: una revisión integradora

SUMMARY: S100 proteins belong group of calcium-binding proteins and are present in physiological intracellular and extracellular regulatory activities, such as cell differentiation, and act in inflammatory and neoplastic pathological processes. Recently, its expressions in the nervous system have been extensively studied, seeking to elucidate its action at the level of the thalamus: A structure of the central nervous system that is part of important circuits, such as somatosensory, behavioral, memory and cognitive, as well as being responsible for the transmission and regulation of information to the cerebral cortex. This article is an integrative review of scientific literature, which analyzed 12 studies present in Pubmed. The analysis showed that the relationship of S100 proteins and the thalamus has been described in neoplastic processes, mental disorders, hypoxia, trauma, stress, infection, Parkinson's disease and epilepsy. In summary, it is possible to conclude that this protein family is relevant as a marker in processes of thalamic injury, requiring further studies to better understand its clinical, preclinical meanings and its prognostic value.

Las proteínas S100 pertenecen al grupo de proteínas fijadoras de calcio y están presentes en actividades reguladoras fisiológicas intracelulares y extracelulares, como la diferenciación celular, y actúan en procesos patológicos inflamatorios y neoplásicos. Recientemente, sus expresiones en el sistema nervioso han sido ampliamente estudiadas, buscando dilucidar su acción a nivel del tálamo: una estructura del sistema nervioso central que forma parte de importantes circuitos, como el somatosensorial, conductual, de memoria y cognitivo, así como además de ser responsable de la transmisión y regulación de la información a la corteza cerebral. Este artículo es una revisión integradora de la literatura científica, que analizó 12 estudios presentes en Pubmed. El análisis mostró que la relación de las proteínas S100 y el tálamo ha sido descrita en procesos neoplásicos, trastornos mentales, hipoxia, trauma, estrés, infección, enfermedad de Parkinson y epilepsia. En resumen, es posible concluir que esta familia de proteínas es relevante como marcador en procesos de lesión talámica, requiriendo más estudios para comprender mejor su significado clínico, preclínico y su valor pronóstico.

Morphology of Echinococcus granulosus protoscolex / Morfología de los Protoescolex de echinococcus granulosus

SUMMARY: Echinococcus granulosus (E. granulosus), is a tapeworm that spreads between intermediate and definitive hosts through the ingestion of fecal matter contaminated with the parasite's eggs. The life cycle consists of differentiation from eggs to oncospheres to embryos, which eventually form cysts in organs like the liver, lungs and others. Within these cysts are protoscolices, an intermediate stage of the parasite which develop into adult tapeworms once they infect their definitive hosts. When these hydatid cysts form in humans, it is known as Cystic Echinococcosis (CE). This disease is treated through surgical excision of the cysts and or chemotherapy with benzimidazole compounds. Understanding the morphology of the intermediate developmental stage of E. granulosus, protoscolex stage, can allow researchers to identify defining structural changes and protein functions that could be used to develop treatment modalities for CE. Unique characteristics in the tegumental surface during the protoescolex stage and proteins associated with cyst fertility have all been identified in previous research studies and bring researchers closer to understanding the underlying mechanisms of E. granulosus development, and consequently, means to disrupt it to achieve better control of the disease.

El Echinococcus granulosus (E. granulosus), es un cestodo que se propaga entre huéspedes intermedios y definitivos a través de la ingestión de materia fecal contaminada con los huevos del parásito. El ciclo de vida consiste en la diferenciación de huevos a oncosferas y embriones, que finalmente forman quistes en órganos como el hígado, los pulmones y otros. Dentro de estos quistes hay protoescólices, una etapa intermedia del parásito que se convierte en su forma adulta (tenia), una vez que infectan a sus huéspedes definitivos. Cuando estos quistes hidatídicos se desarrollan en seres humanos, se les conoce como equinococosis quística (EC). Esta enfermedad se trata mediante la extirpación quirúrgica de los quistes o la quimioterapia con compuestos benzimidazólicos. La comprensión de la morfología de la etapa de desarrollo intermedia del E. granulosus y la etapa de protosclex, puede permitir a los investigadores identificar cambios estructurales definidos y funciones de proteínas que podrían usarse para desarrollar modalidades de tratamiento para la CE. Las características únicas en la superficie tegumentaria durante la etapa de protoescolex y las proteínas asociadas con la fertilidad del quiste se han identificado en estudios de investigación anteriores y acercan a los investigadores a la comprensión de los mecanismos subyacentes del desarrollo del E. granulosus y, en consecuencia, los medios para interrumpirlo para lograr un mejor control de la enfermedad.

Preliminary study on the effect of Echinococcus multilocaris on phenotypic transformations of glucose metabolism and polarization types in macrophages / 中国血吸虫病防治杂志

Objective To investigate the effects of Echinococcus multilocularis on the phenotypic transformations of glucose metabolism, polarization types and inflammatory responses in macrophages, so as to provide insights into elucidation of echinococcosis pathogenesis. Methods Bone marrow cells were isolated from C57BL/6J mice at ages of 6 to 8 weeks, and induced into bone marrow-derived macrophages (BMDMs) with mouse macrophage colony-stimulating factor (M-CSF), which served as controls (BMDMs-M0). BMDMs-M0 induced M2 macrophages by interleukin-4 for 24 hours served as the IL-4 induction group, and BMDMs-M0 co-cultured with 2.4 ng/mL E. multilocularis cystic fluid (CF) served as the BMDM-CF co-culture group, while BMDMs-M0 co-cultured with E. multilocularis protoscolex (PSC) at a ratio of 500:1 served as the BMDM-PSC co-culture group. The types of polarization of BMDMs co-cultured with E. multilocularis CF and PSC were analyzed using flow cytometry, and the expression of macrophage markers, inflammatory factors, and glucose metabolism-related enzymes was quantified using fluorescent quantitative real-time PCR (qPCR) and Western blotting assays. Results There were significant differences among the four groups in terms of Arginase-1 (Arg1) (F = 1 457.00, P < 0.000 1), macrophages-derived C-C motif chemokine 22 (Ccl22) (F = 22 203.00, P < 0.000 1), resistin-like α (Retnla) (F = 151.90, P < 0.000 1), inducible nitric oxide synthase (iNOS) (F = 107.80, P < 0.001), hexokinase (HK) (F = 9 389.00, P < 0.000 1), pyruvate kinase (PK) (F = 641.40, P < 0.001), phosphofructokinase 1 (PFK1) (F = 43.97, P < 0.01), glucokinase (GK) (F = 432.50, P < 0.000 1), pyruvate dehydrogenase kinases1 (PDK1) (F = 737.30, P < 0.000 1), lactic dehydrogenase (LDH) (F = 3 632.00, P < 0.000 1), glucose transporter 1 (GLUT1) (F = 532.40, P < 0.000 1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (F = 460.00, P < 0.000 1), citrate synthase (CS) (F = 5 642.00, P < 0.01), glycogen synthase1 (GYS1) (F = 273.30, P < 0.000 1), IL-6 (F = 1 823.00, P < 0.000 1), IL-10 (F = 291.70, P < 0.000 1), IL-1β (F = 986.60, P < 0.000 1), and tumor necrosis factor (TNF)-α (F = 334.80, P < 0.000 1) and transforming growth factor (TGF)-β mRNA expression (F = 163.30, P < 0.001). The proportion of M2 macrophages was significantly higher than that of M1 macrophages in the BMDM-PSC co-culture group [(22.87% ±1.48%) vs. (1.70% ±0.17%); t = 24.61, P < 0.001], and the proportion of M2 macrophages was significantly higher than that of M1 macrophages in the BMDM-CF co-culture group [(20.07% ±0.64%) vs. (1.93% ±0.25%); t = 45.73, P < 0.001]. The mRNA expression of M2 macrophages markers Arg1, Ccl22 and Retnla was significantly higher in the BMDM-CF and BMDM-PSC co-culture groups than in the control group (all P values < 0.01), and no significant difference was seen in the mRNA expression of the M1 macrophage marker iNOS among the three groups (P > 0.05), while qPCR assay quantified higher mRNA expression of key glycolytic enzymes HK, PK and PFK, as well as inflammatory factors IL-10, IL-1β, TNF-α and TGF-β in the BMDM-CF and BMDM-PSC co-culture groups than in the control group (all P values < 0.01). Western blotting assay determined higher HK, PK and PFK protein expression in the BMDM-PSC co-culture group than in the control group (all P values < 0.05), and qPCR quantified higher GLUT1, GAPDH and IL-6 mRNA expression in the BMDM-CF co-culture group than in the control group (all P values < 0.05), while higher HK, PK and PFK protein and mRNA expression (all P values < 0.01), as well as lower IL-6 and TNF-α and higher TGF-β mRNA expression (both P values < 0.05) was detected in the IL-4 induction group than in the control group. Glycolytic stress test showed no significant difference in the extracellular acidification rate (ECAR) of mouse BMDM among the control group, IL-4 induction group and BMDM-PSC co-culture group (F = 124.4, P < 0.05), and a higher ECAR was seen in the BMDM-PSC co-culture group and a lower ECAR was found in the IL-4 induction group than in the control group (both P values < 0.05). Conclusions Treatment of E. multilocularis CF or PSC mainly causes polarization of BMDM into M2 macrophages, and phenotypic transformation of glucose metabolism into high-energy and high-glycolytic metabolism, and affects inflammatory responses in BMDM.

Estudio morfocuantitativo de protoescólices y otras estructuras fundamentales de Echinococcus granulosus en hospederos humanos / Morphoquantitative study of protoscolices and other fundamental structures of Echinococcus granulosus in human hosts

La equinococosis quística (EQ) a pesar de ser una enfermedad endémica en diversos lugares del planeta, presenta pocos estudios morfológicos y cuantitativos de las estructuras fundamentales del Echinococcus granulosus en humanos, en especial de los protoescólices. El objetivo de este estudio fue analizar morfocuantitativamente protoescólices y otras estructuras fundamentales de E. granulosus obtenidos de hospederos humanos. Estudio de corte transversal. Se estudiaron 8 especímenes de EQ hepática humana, aplicando un muestreo no probabilístico por conveniencia. Se evaluó las capas quísticas, el líquido y la arenilla hidatídica. Las capas fueron fijadas en formaldehido tamponado a 10 % y procesadas para su inclusión en paraplast. Se realizaron cortes de 5 µm de grosor y fueron teñidas con H-E para su análisis con microscopía óptica. El líquido y arenilla fueron centrifugados y al sedimento obtenido se le realizó análisis directo para determinar las medidas morfométricas de los protoescólices y de los ganchos grandes y pequeños. Se utilizó estadística descriptiva. De los 8 quistes estudiados, 6 eran quistes univesicular, uno multivesicular y un quiste abscedado, cuyas capas laminada y germinativa se encontraban bien definidas. Las vesículas prolígeras presentaban forma redondeada con protoescólices en su interior. Los protoescólices invaginados presentaron un largo y ancho promedio de 140,8 ± 34,3 µm y 106,2 ± 29,5 µm, respectivamente; y los desarrollados un largo de 237,2 ± 53,0 µm y ancho de 128,7 ± 32,0 µm. Los ganchos rostelares presentaron contornos suaves distribuidos en dos filas regulares. El promedio del largo total de los ganchos grandes y pequeños fue 20,1 ± 2,7 µm; el promedio del ancho total fue 7,4 ± 1,2 µm. En conclusión, las características morfocuantitativas de los ganchos de E. granulosus en humanos, son distintos a otras especies hospederas intermediarias y de otros Echinococcus spp. Es posible que el abandono del estado de resistencia de los protoescólices invaginado hasta su desarrollo genere implicancias epidemiológicas de interés.

SUMMARY: Although cystic echinococcosis (CE) is an endemic disease in several parts of the world, few morphological and quantitative studies of the fundamental structures of Echinococcus granulosus in humans, especially protoscolices. The aim of this study was to perform a morphoquantitative analysis of protoescolex and other fundamental structures of E. granulosus from human hosts. Cross- sectional study. Eight human hepatic EQ specimens were studied, applying non-probabilistic convenience sampling. Cystic layers, fluid and hydatid grit were evaluated. The layers were fixed in 10% buffered formaldehyde and processed for embedding in paraplast. Slices of 5 µm thickness were made and stained with H-E for light microscopic analysis. The liquid and grit were centrifuged and the sediment obtained was analyzed directly to determine the morphometric measurements of the protoscolices and the large and small hooks. Descriptive statistics were used. Of the 8 cysts studied, 6 were univesicular cysts, one multivesicular and one abscessed cyst, whose lamellar and germinative layers were well defined. The proligerous vesicles had a rounded shape with protoscolices inside. The invaginated protoscolices had an average length and width of 140.8 ± 34.3 µm and 106.2 ± 29.5 µm, respectively; and the developed ones had a length of 237.2 ± 53.0 µm and width of 128.7 ± 32.0 µm. The rostellar hooks presented smooth contours distributed in two regular rows. The average total length of the large and small hooks was 20.1 ± 2.7 µm; the average total width was 7.4 ± 1.2 µm. In conclusion, the morphoquantitative characteristics of E. granulosus hooks in humans are distinct from other intermediate host species and from other Echinococcus spp. It is possible that the abandonment of the resistance state of the invaginated protoscolices until their development generates epidemiological implications of interest.

Mesenchymal stem cell calcification induced by protoscolex of two species of Echinococcus: A differential analysis / 中国组织工程研究

BACKGROUND: The growth pattern of Echinococcus granulosus is different from that of Echinococcus alveolaris. Hepatic echinococcosis can form a complete fibrous calcified cyst wall, while hepatic alveolar echinococcosis can grow infiltratively and cannot form a complete fibrous calcified cyst wall. Bone marrow mesenchymal stem cells (BMSCs) are involved in the formation of calcified wall of hydatidosis, but the calcification characteristics of Echinococcus granulosus and Echinococcus alveolaris are different and the role of BMSCs is still unclear. OBJECTIVE: To compare the effects of Echinococcus granulosus and Echinococcus alveolaris on the calcification of BMSCs and to preliminarily investigate the formation mechanism of echinococcosis calcifications. METHODS: BMSCs of C57BL/6 mice were extracted, cultured and identified, followed by co-culture with the protoscolex of Echinococcus granulosus (BMSC+CE group) and Echinococcus alveolaris (BMSC+AE group), respectively. BMSCs cultured alone were used as control group. After 1, 4, and 7 days of co-culture, alkaline phosphatase activity was detected by a microplate reader, the expression of BMP2 and RUNX2 mRNA was detected by RT-q PCR, and the expression of BMP2, RUNX2 and phosphorylated Smad1/5/8 (P-Smad1/5/8) proteins was detected by western blot assay. RESULTS AND CONCLUSION: (1) The alkaline phosphatase activity of the BMSC+CE group and BMSC+AE group was significantly higher than that of the control group at 1 and 4 days after culture (P < 0.05), and the alkaline phosphatase activity of the BMSC+CE group was significantly higher than that of BMSC+AE group (P < 0.05). (2) Western blot results showed that the expression of BMP2, RUNX2, and P-Smad1/5/8 protein in the BMSC+CE group and BMSC+AE group was significantly higher than that in the control group at 1 and 4 days after culture (P < 0.05), while the expression of BMP2, RUNX2, and P-Smad1/5/8 protein in the BMSC+CE group was significantly higher than that in the BMSC+AE group (P < 0.05). (3) RT-qPCR results showed that the expression of BMP2 and RUNX2 mRNA in the BMSC+CE group was significantly higher than that in the control group at 1, 4 and 7 days after culture (P < 0.05), and was significantly higher than that in the BMSC+AE group at 4 and 7 days after culture (P < 0.05). The expression of RUNX2 mRNA in the BMSC+AE group was significantly higher than that in the control group at 1, 4, and 7 days after culture (P < 0.05). (4) To conclude, co-culture of the protoscolex of Echinococcus alveolaris and BMSCs promotes the expression of alkaline phosphatase and RUNX2 in BMSCs by up-regulating BMP-Smad1/5/8 pathway. At the later stage of co-culture, the effect of Echinococcus alveolaris on BMSCs calcification is significantly weakened, while the effect of Echinococcus granulosus on BMSCs calcification remains unchanged, suggesting that this mechanism may be related to the different growth patterns of two kinds of hydatids.

In-vitro evaluation of Nigella sativa and Punica granatum effect on protoscolices of hydatid cysts / Avaliação in vitro do efeito de Nigella sativa e Punica granatum em protoescólices de cistos hidáticos

Abstract Cystic echinococcosis (CE) are commonly found in the liver and lungs of affected hosts. The treatment approach is usually surgical, or giving drugs in conjunction before surgery to kill protoscolices, to avoid anaphylactic shock from leakage of hydatid fluid into the peritoneum and to decrease opportunities for recurrences. The present study was to evaluate the in vitro scolicidal efficacy of hydroalcoholic extract of Punica granatum peel and Nigella sativa, on the protoscolices of CE that collected from the lungs of infected camels. Different concentrations of extracts with different exposure times were used and a viability assay was applied to measure the scolicidal effect. N. sativa showed its highest scolicidal efficacy at 100 mg/mL and 10 mg/mL concentrations after 30 and 60 min. P. granatum peel extract showed its maximum scolicidal efficacy at 100 mg/mL concentration after 120 min. All experiments of the current study revealed that the extracts of both N. sativa and P. granatum had a scolicidal effects on the protoscolices of camel hydatid cysts. It could be concluded that N. sativa extract is more potent than P. granatum peel extract regarding scolicidal effect, but the efficacies of both extracts were of moderate significant correlation to exposure time and concentrations.

Resumo Os cistos hidáticos (equinococose cística, CE) são comumente encontrados no fígado e nos pulmões dos hospedeiros afetados. A abordagem do tratamento geralmente é cirúrgica, e algumas drogas são administradas em conjunto antes da cirurgia para matar protoscólices e evitar choque anafilático devido ao vazamento de fluido hidático no peritônio e diminuir as oportunidades de recorrência. O presente estudo foi avaliar a eficácia in-vitro do extrato hidroalcoólico de casca de Punica granatum e Nigella sativa, sobre os protoescólices de cistos hidáticos, que foram coletados dos pulmões de camelos infectados. Concentrações dos extratos com diferentes tempos de exposição foram utilizados e um ensaio de viabilidade foi aplicado para medir o efeito escolicida. A N. sativa apresentou sua maior eficácia escolicida nas concentrações de 100 mg/mL e 10 mg/mL após 30 e 60 min. O extrato de casca de P. granatum mostrou sua máxima eficácia escolicida na concentração de 100 mg/mL após 120 min. Todos os experimentos do presente estudo revelaram que os extratos de N. sativa e P. granatum tiveram efeito escolicida dependente da dose e do tempo nos protoescólices dos cistos hidáticos de camelo. Pode-se concluir que o extrato de N. sativa é mais potente que o extrato de casca de P. granatum em relação ao efeito escolicida, mas a eficácia de ambos os extratos foi de correlação significativa moderada com o tempo de exposição e as concentrações.

Evaluating the effect of Myrtus communis on programmed cell death in hydatid cyst protoscolices

OBJECTIVE@#To evaluate the possible involvement of programmed cell death strategy in hydatid cyst protoscolices following treatment with Myrtus communis (M. communis) as an herbal medicine.@*METHODS@#Protoscolices were aseptically collected from sheep liver hydatid cysts. Evaluating the effect of M. communis extract on programmed cell death and increased activity of caspases 3, 8, and 9 in hydatid cyst protoscolices was conducted by treating the protoscolices with different concentration (5, 50, and 100 mg/mL) of M. communis extract at 37 °C and 5% CO for 4 h by using the Bradford test and ELISA commercial kits.@*RESULTS@#The extract of M. communis at all concentrations led to initiation of programmed cell death in protoscolices and this effect, was only significant at 50 and 100 mg/mL concentrations, compared to the negative control (P < 0.05). Also, the activity of caspases 3, 8, and 9 in hydatid cyst protoscolices, was shown that the extract at all 3 concentrations could only increase the activity of caspases 3 and 9. Moreover, a significant increase in the activity of caspase 3 was only observed at concentrations 50 and 100 mg/mL by 37.00% and 66.19% while a significant increase in the activity of caspase 9 at the same concentrations was observed by 20.89% and 63.67%, respectively (P < 0.05).@*CONCLUSIONS@#The extract of M. communis at different concentrations could increase the activity of caspases 3 and 9 and caused programmed cell death in hydatid cyst protoscolices however, this effect was significant at high concentrations of the extract.

Evaluating the effect of Myrtus communis on programmed cell death in hydatid cyst protoscolices

Objective To evaluate the possible involvement of programmed cell death strategy in hydatid cyst protoscolices following treatment with Myrtus communis (M. communis) as an herbal medicine. Methods Protoscolices were aseptically collected from sheep liver hydatid cysts. Evaluating the effect of M. communis extract on programmed cell death and increased activity of caspases 3, 8, and 9 in hydatid cyst protoscolices was conducted by treating the protoscolices with different concentration (5, 50, and 100 mg/mL) of M. communis extract at 37 °C and 5% CO

Effect of protoscolex on T subsets in mice spleen cells in vitro / 中国免疫学杂志

Objective:To observe the effect of protoscolex on Th subsets and correlative cytokine in mice spleen cells in vitro.Methods:Co-culture spleen cells from BALB/c mice with protoscolices,then IL-4,IFN-γand TGF-βproduction in cell culture supernatants were analyzed by ELISA.The percentage of Th subsets were detected by Flow Cytometry analysis.Results:Secretion levels of IL-4 and TGF-βwere significantly increased in spleen cells at different time point in co-culture system with protoscolices.Ratios of Th2 and Treg cells were also significantly increased in co-culture system at different time points than the control groups.However,there was no statistical significance for ratio of Th1 cells at different time points.Conclusion:The protoscolex can increase the ratios of Th2 cells and Treg cells from spleen cells.Secretion levels of IL-4 and TGF-βwere also increased in spleen cells co-cultured with protosco-lices.The results suggest that these Th cell subsets play a role in the immune escape of the hydatid disease.

Molecular Cloning and Characterization of a P38-Like Mitogen-Activated Protein Kinase from Echinococcus granulosus

Cystic echinococcosis (CE) treatment urgently requires a novel drug. The p38 mitogen-activated protein kinases (MAPKs) are a family of Ser/Thr protein kinases, but still have to be characterized in Echinococcus granulosus. We identified a 1,107 bp cDNA encoding a 368 amino acid MAPK protein (Egp38) in E. granulosus. Egp38 exhibits 2 distinguishing features of p38-like kinases: a highly conserved T-X-Y motif and an activation loop segment. Structural homology modeling indicated a conserved structure among Egp38, EmMPK2, and H. sapiens p38α, implying a common binding mechanism for the ligand domain and downstream signal transduction processing similar to that described for p38α. Egp38 and its phosphorylated form are expressed in the E. granulosus larval stages vesicle and protoscolices during intermediate host infection of an intermediate host. Treatment of in vitro cultivated protoscolices with the p38-MAPK inhibitor ML3403 effectively suppressed Egp38 activity and led to significant protoscolices death within 5 days. Treatment of in vitro-cultivated protoscolices with TGF-β1 effectively induced Egp38 phosphorylation. In summary, the MAPK, Egp38, was identified in E. granulosus, as an anti-CE drug target and participates in the interplay between the host and E. granulosus via human TGF-β1.

Construction of In Vivo Fluorescent Imaging of Echinococcus granulosus in a Mouse Model

Human hydatid disease (cystic echinococcosis, CE) is a chronic parasitic infection caused by the larval stage of the cestode Echinococcus granulosus. As the disease mainly affects the liver, approximately 70% of all identified CE cases are detected in this organ. Optical molecular imaging (OMI), a noninvasive imaging technique, has never been used in vivo with the specific molecular markers of CE. Thus, we aimed to construct an in vivo fluorescent imaging mouse model of CE to locate and quantify the presence of the parasites within the liver noninvasively. Drug-treated protoscolices were monitored after marking by JC-1 dye in in vitro and in vivo studies. This work describes for the first time the successful construction of an in vivo model of E. granulosus in a small living experimental animal to achieve dynamic monitoring and observation of multiple time points of the infection course. Using this model, we quantified and analyzed labeled protoscolices based on the intensities of their red and green fluorescence. Interestingly, the ratio of red to green fluorescence intensity not only revealed the location of protoscolices but also determined the viability of the parasites in vivo and in vivo tests. The noninvasive imaging model proposed in this work will be further studied for long-term detection and observation and may potentially be widely utilized in susceptibility testing and therapeutic effect evaluation.

In Vitro Effects of SB202190 on Echinococcus granulosus

Spillage of cyst contents during surgical operation is the major cause of recurrence after hydatid cyst surgery. Instillation of a scolicidal agent into a hepatic hydatid cyst is the most commonly employed measure to prevent this complication. SB202190 is a pyridinyl imidazole derivative and is known to be a specific inhibitor of p38 MAPK. In the present study, the scolicidal effect of SB202190 was investigated. Freshly isolated Echinococcus granulosus protoscolices were subjected to SB202190 treatment (10, 20, 40, and 80 microM), and the effects on parasite viability were monitored by trypan blue staining. Corresponding effects were visualized by scanning and transmission electron microscopy. Dose-dependent protoscolex death within a few days of SB202190 treatment was observed. Although the in vitro scolicidal effect of SB202190 was satisfactory, the in vivo efficacy of this drug and also possible side effects remain to be further investigated.

In Vitro Effects of Some Herbs Used in Egyptian Traditional Medicine on Viability of Protoscolices of Hydatid Cysts

The present work evaluated the effects of alcoholic extracts of salvia (Salvia officinalis), thyme (Thymus vulgaris), and 2 pure compounds (thymol and menthol) on the viability of Echinococcus granulosus protoscolices in vitro. Four different concentrations of each extract (2,500, 1,500, 1,000, and 500 microg/ml) and 3 different concentrations each of thymol and menthol (50, 10, and 1 microg/ml) were used. Concentration of 2,500 microg/ml of both extracts showed a significant protoscolicidal activity on the 6th day. Complete loss of viability of protoscolices occurred with 500 microg/ml concentration of both extracts at day 6 and day 7 post-treatment (PT), respectively. Pure compounds, i.e., menthol and thymol, showed potent effects with 50 microg/ml concentration at day 2 and day 5 PT, respectively. These effects were compared with those of albendazole sulfoxide (800 microg/ml), a commonly used treatment drug for hydatidosis. Krebs-Ringer solution and the hydatid cystic fluid at a ratio of 4:1 was a good preservative solution which kept the protoscolices viable for 15 days.

An Imported Case of Echinococcosis of the Liver in a Korean Who Traveled to Western and Central Europe

Echinococcus granulosus, an intestinal tapeworm of dogs and other canids, infects humans in its larval stage and causes human echinococcosis or hydatid disease. In the Republic of Korea, 31 parasite-proven human echinococcosis cases have been reported, most of which were imported from the Middle East. We recently examined a 61-year-old Korean man who had a large cystic mass in his liver. ELISA was negative for tissue parasitic infections, including echinococcosis, cysticercosis, paragonimiasis, and sparganosis. The patient underwent surgery to remove the cyst, and the resected cyst was processed histopathologically for microscopic examinations. In sectioned cyst tissue, necrotizing protoscolices with disintegrated hooklets of E. granulosus were found. In some areas, only freed, fragmented hooklets were detected. The patient had traveled to western and central Europe in 1996, and had no other history of overseas travel. We report our patient as a hepatic echinococcosis case which was probably imported from Europe.

Dot-Elisa for evaluation of hydatid cyst wall, protoscoleces and hydatid cyst fluid antigens in the serodiagnosis of cystic echinococcosis / Dot-Elisa para avaliação da parede do cisto hidático, protoscoleces e antígenos do fluido do cisto hidático no sorodiagnóstico da equinococose cística

The aim of the present study is to evaluate cyst wall and protoscolex as an alternate source of antigen in serodiagnosis of cystic echinococcosis (CE). A total of 90 blood samples, 30 each of confirmed CE cases, disease controls and healthy controls were collected. Dot-ELISA using cyst wall, protoscolex and cyst fluid were used to demonstrate anti-hydatid antibodies. The sensitivity of Dot-ELISA using cyst wall, protoscolex and cyst fluid was 96.66 percent, 86.66 percent and 93.33 percent respectively and the specificity of the assay was 70 percent for Dot-ELISA using cyst fluid, protoscolex and cyst wall antigens. Results of the present study show that cyst wall and protoscolex can also be an useful source of antigen in detection of hydatid antibodies in the serodiagnosis of CE.

O propósito do presente trabalho é avaliar a parede cística e protoscolex como fontes alternativas de antígeno no sorodiagnóstico de equinococose cística (CE). De um total de 90 amostras de sangue, foram coletadas 30 de casos CE confirmados, 30 de controles de doença e 30 controles saudáveis. Dot-Elisa usando parede cística, protoscolex e fluido cístico foi utilizada para demonstrar anticorpos anti-hidáticos. A sensitividade de Dot-Elisa usando parede cística, protoscolex e fluido cístico foi de: 96,66 por cento, 86,66 por cento e 93,33 por cento respectivamente e a especificidade do ensaio de 70 por cento para Dot-Elisa usando fluido cístico, protoscolex e antígeno da parede cística. Resultados do presente estudo mostram que parede cística e protoscolex podem ser fontes úteis de antígeno na detecção de anticorpos hidáticos no sorodiagnóstico do CE.

In Vitro Observation on Albendazole Sulfoxide and its Enantiomers against Echinococcus granulosus Protoscolex / 中国寄生虫学与寄生虫病杂志

Objective To investigate in vitro anti-hydatid efficacy on Echinococcus granulosus protoscolex(EgPSC) by using albendazole sulfoxide(ASOX) and its two enantiomeric antipodes, L-ASOX and D-ASOX.Methods Eg protoscoleces were divided into eight groups and cultured in the DMEM culture media under two concentrations(50 ?g/ml and 100 ?g/ml) of ASOX, L-ASOX and D-ASOX respectively.The appropriate controls included(i) a culture containing an equal amount of DMSO and(ii) a culture medium alone.The mortality of EgPSC in each group was daily counted until 100% EgPSC death in some groups.Results Significant difference of EgPSC mortality was found among the three drugs with various concentrations compared to control group(P0.05), but between ASOX group and L-ASOX group(P

In vitro Observation on the Apoptosis Induced by H_2O_2 in Protoscolex of Echinococcus granulosus / 中国寄生虫学与寄生虫病杂志

Objective To explore the apoptosis induced by hydrogen peroxide ( H2O2) in protoscolex of Echinococcus granulosus. Methods Protoscoleces were cultured in vitro, and used for the experiment in 2 groups: RPMI 1640 medium and RPMI 1640 medium added with glutamine. They were then treated with different concentrations of H2O2 to induce apoptosis. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay (TUNEL) was employed to observe the apoptosis. Protein expression of caspase-1, caspase-3 and Fas was detected by SP immunohistochemical technique, stained with DAB restained with hematoxylin. A yellow or brown color nucleus revealed positive apoptosis cells in protoscolex, a brown reaction product in cytoplasm showed positive cells of caspase-1 and caspase-3, and brown cell membrane and cytoplasm revealed Fas product; otherwise it was judged as negative. According to the percentage of positive cells in a protoscolex, the expression level was divided as 4 grades. The percentage of less than 5% was regarded as "-", 5%-25% as "+", 26%-50% as "++", more than 50% as "+++". The experiments were repeated 2 times with controls. Results In RPMI 1640 group, positive TUNEL was found in the protoscolex induced by 1 mmol/L H2O2 inducing for 12 hours. Induced by 1 mmol/L H2O2 for 4 h, the "+ -++" expression rate of caspase-1 and caspase-3 in the protoscoleces was 86.6% and 77.8% , and for 8 h, 86.6% and 80.0% respectively, a significant increase in comparison to the control (P