Application of an ELISA test using Schistosoma bovis adult worm antigens in travellers and immigrants from a schistosomiasis endemic area and its correlation with clinical findings

[EN] We have recently evaluated an ELISA for the diagnosis of human schistosomiasis using S. bovis adult worm antigens (AWA Sb), showing a sensitivity of 94% and a specificity of 97% for patients diagnosed by egg detection. Nevertheless, the comparison of this AWA Sb ELISA with direct parasitological findings as the gold standard could introduce a Selection bias, due to the well-known lack of sensitivity of direct methods in the detection of acute schistosomiasis and of low burden infections. The objective of the present work is to compare it with parasitological methods and commercial indirect haemagglutination test using S. mansoni antigens (WA Sm IHA) in 254 immigrants and travellers with different clinical settings; in addition, to find specific bands in the EITB of different phases of schistosomiasis. The AWA Sb ELISA showed 72% of seropositivity in patients with Katayama fever, while patients with eosinophilia and genito-urinary complaints showed 27% and 93%, respectively. The diagnosis yield was globally higher than direct egg detection or WA Sm IHA test with regard to the clinical setting. Finally, the utilization of EITB with S. bovis AWA permits the confirmation of diagnosis in chronic and acute phases of the disease

EducaFarma 8.0

Memoria ID-033 . Ayudas de la Universidad de Salamanca para la innovación docente, curso 2019-2020

Malaria proteomic research

Malaria is one of the main infectious diseases in the world, particularly in tropical and subtropical areas. Among the species that can cause this disease, in recent years P. vivax has been growing due to its own attributes, which make it highly difficult to eradicate. This study was focused on analyzing and identifying the proteome of P. vivax during the blood stage through a mass spectrometry analysis (LC-MS-MS). Results allowed us to identify 743 proteins, of which 522 never had been described before. Furthermore, the comparison of the expression of these proteins with P. vivax transcriptional profile allowed us to corroborate the adaptive change in the P. vivax VCG-1 strain transcriptome, previously described.La malaria sigue siendo una de las principales enfermedades transmisibles del planeta, especialmente en áreas tropicales y subtropicales. Dentro de las diversas especies causantes de esta enfermedad, en los últimos años ha ganado relevancia el estudio de P. vivax debido a sus características propias, que la hacen especialmente difícil de erradicar. En el presente estudio, nos proponemos analizar e identificar las proteínas de la fase hemática de P.vivax mediante cromatografía líquida asociada a espectrometría de masas en tándem (LC-MS-MS). Los resultados del estudio nos permitieron identificar 743 proteínas, de las que 522 no habían sido previamente descritas. Además, la comparación del perfil de expresión de estas proteínas con el perfil transcripcional de P. vivax nos permitió corroborar lo descrito en estudios anteriores: el cambio adaptativo en el perfil transcripcional de la cepa VCG-1 de P. vivax

Detection of Schistosoma mansoni-derived DNA in human urine samples by loopmediated isothermal amplification (LAMP)

[EN]Background Schistosoma mansoni is the main species causing hepatic and intestinal schistosomiasis in Sub-Saharan Africa, and it is the only species in South America. Adult stages of the parasite reside in the mesenteric venous plexus of infected hosts, and eggs are shed in feces. Collecting patient stool samples for S. mansoni diagnostic purposes is difficult in large-scale field trials. Urine samples would be an alternative approach for molecular S. mansoni detection since they have several advantages over stool samples, including better handling, management and storage. Additionally, loop-mediated isothermal amplification (LAMP) technology is a powerful molecular diagnostic tool for infectious diseases, particularly under field conditions in developing countries. The present study aimed to assess the effectiveness of our previously developed LAMP assay (SmMIT-LAMP) for S. mansoni-specific detection in clinical urine samples. Methodology/Principal findings The sensitivity of SmMIT-LAMP in urine was established in simulated fresh human urine samples artificially spiked with genomic DNA from S. mansoni. LAMP for 120 min instead of 60 min improved the sensitivity, reaching values of 0.01 fg/μL. A set of well-defined frozen stored human urine samples collected from Sub-Saharan immigrant patients was selected from a biobank to evaluate the diagnostic validity of SmMIT-LAMP. The set included urine samples from patients with microscopy-confirmed infections with S. mansoni, S. haematobium and other nonschistosome parasites, as well as urine samples from patients with microscopy-negative eosinophilia without a confirmed diagnosis. The SmMIT-LAMP was incubated for 60 and 120 min. A longer incubation time was shown to increase the LAMPpositive results in patient urine samples. We also tested urine samples from mice experimentally infected with S. mansoni, and LAMP-positive results were obtained from the third week after infection. A real-time LAMP assay was also performed with three individual urine samples. Conclusions/Significance The SmMIT-LAMP could effectively detect S. mansoni DNA in mouse urine samples and produced promising results for human clinical samples. The detection of S. mansoni DNA in mouse urine samples from the third week after infection indicates that early diagnosis of active S. mansoni infection is possible using urine as a source of DNA. Further studies are still needed, but our method could be used as a promising molecular tool applicable to urine samples to diagnose human intestinal schistosomiasis caused by S. mansoni

Gene expression analyses determine two different subpopulations in KIT-negative GIST-like (KNGL) patients

Introduction: there are limited findings available on KIT-negative GIST-like (KNGL) population. Also, KIT expression may be post-transcriptionally regulated by miRNA221 and miRNA222. Hence, the aim of this study is to characterize KNGL population, by differential gene expression, and to analyze miRNA221/222 expression and their prognostic value in KNGL patients. Methods: KIT, PDGFRA, DOG1, IGF1R, MIR221 and MIR222 expression levels were determined by qRT-PCR. We also analyzed KIT and PDGFRA mutations, DOG1 expression, by immunohistochemistry, along with clinical and pathological data. Disease-free survival (DFS) and overall survival (OS) differences were calculated using Log-rank test. Results: hierarchical cluster analyses from gene expression data identified two groups: group I had KIT, DOG1 and PDGFRA overexpression and IGF1R underexpression and group II had overexpression of IGF1R and low expression of KIT, DOG1 and PDGFRA. Group II had a significant worse OS (p = 0.013) in all the series, and showed a tendency for worse OS (p = 0.11), when analyzed only the localized cases. MiRNA222 expression was significantly lower in a control subset of KIT-positive GIST (p < 0.001). OS was significantly worse in KNGL cases with higher expression of MIR221 (p = 0.028) or MIR222 (p = 0.014). Conclusions: we identified two distinct KNGL subsets, with a different prognostic value. Increased levels of miRNA221/222, which are associated with worse OS, could explain the absence of KIT protein expression of most KNGL tumors

Inicio de la gestión de la Farmacia de AUSAF (Aula de Atención Farmacéutica de la Universidad de Salamanca)

Memoria ID-204. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2018-2019

EducaFarma 7.0

Memoria ID-013. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2018-2019

EducaFarma 9.0

Memoria ID-020 Ayudas de la Universidad de Salamanca para la innovación docente, curso 2020-2021

Malignant bone tumors (other than Ewing's) : clinical practice guidelines for diagnosis, treatment and follow-up by Spanish Group for Research on Sarcomas (GEIS)

Primary malignant bone tumors are uncommon and heterogeneous malignancies. This document is a guideline developed by the Spanish Group for Research on Sarcoma with the participation of different specialists involved in the diagnosis and treatment of bone sarcomas. The aim is to provide practical recommendations with the intention of helping in the clinical decision-making process. The diagnosis and treatment of bone tumors requires a multidisciplinary approach, involving as a minimum pathologists, radiologists, surgeons, and radiation and medical oncologists. Early referral to a specialist center could improve patients' survival. The multidisciplinary management of osteosarcoma, chondrosarcoma, chordoma, giant cell tumor of bone and other rare bone tumors is reviewed in this guideline. Ewing's sarcoma will be the focus of a separate guideline because of its specific biological, clinical and therapeutic features. Each statement has been accompanied by the level of evidence and grade of recommendation on the basis of the available data. Surgical excision is the mainstay of treatment of a localized bone tumor, with various techniques available depending on the histologic type, grade and location of the tumor. Chemotherapy plays an important role in some chemosensitive subtypes (such as high-grade osteosarcoma). In other subtypes, historically considered chemoresistant (such as chordoma or giant cell tumor of bone), new targeted therapies have emerged recently, with a very significant efficacy in the case of denosumab. Radiation therapy is usually necessary in the treatment of chordoma and sometimes of other bone tumors