Schistosomiasis is one of the most important helminthic parasitic infections in the world, with over 700 million people at risk of infection. Species of Schistosoma have a complex life cycle involving the infection of freshwater snails before infecting their mammalian definitive host.  https://www.selleckchem.com/products/Rapamycin.html  Taking about 130,000 lives per annum, S. mansoni is the major cause of intestinal schistosomiasis worldwide. Within Biomphalaria glabrata snails, asexual replication of the parasite gives rise to cercariae larvae. Cercariae actively penetrate the host's skin to complete their life cycle and eventually transform into adult worms. If left untreated, intestinal schistosomiasis can lead to peripheral destruction of the portal vein system, gastric hemorrhage from esophageal varices, as well as hepatic failure. Mass spectrometry (MS) is the method of choice for proteomics analysis. The bottom-up proteomics approach-also known as "shotgun proteomics"-typically includes a protein extraction and solubilization step followed by proteolytic digestion and tandem MS (MS/MS) analysis. Proteins are later identified by peptide de novo sequencing upon MS and MS/MS spectra of digest peptides. In this chapter, we introduce an analytical workflow for proteome profiling of S. mansoni cercariae using bottom-up proteomics. The cercariae were isolated and lysed. Proteins were then extracted, enzymatically digested, and subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Proteins were identified using MaxQuant software. Cercariae are the first life stage of the parasite S. mansoni which humans encounter, and conducting proteomic analysis on this life cycle stage can shed light on possible drug or vaccine candidates to help disable the parasite's ability to infect or arm the immune system for parasite clearance.Schistosomes are deadly pathogens responsible for the neglected tropical disease schistosomiasis. The parasite's virulence is aided by a skin-like tissue called the tegument. The study of the tegument is hampered by a lack of tools suitable for visualizing the tissue. Here we describe a novel methodology employing fluorophore-conjugated dextrans that allows specific fluorescent labeling of the tegument and that is compatible with downstream fluorescence-labeling techniques including phalloidin labeling, RNA FISH, and immunofluorescence.Individual developmental stages of blood fluke Schistosoma mansoni excrete or secrete a different set of molecules. Here we describe optimized protocols for collection of excretory/secretory products (E/S products) from cercariae, schistosomula, adult worms, and eggs. These E/S products are essential for successful parasitism functioning at the host-parasite interface, enabling invasion into the host and contributing to the survival of the parasite by modulation of host physiology and immune responses. Collection of sufficient amounts of E/S products is required for detailed research of these processes.In situ hybridization is a tool for evaluation of gene expression within tissues or single cells. This protocol describes optimized sensitive fluorescence detection of gene transcripts (mRNAs) in semithin sections of Schistosoma mansoni adult worms using specifically designed and labeled RNA probes. Due to improved methodologies in tissue preservation, sectioning, amplification of fluorescent signal, and prehybridization tissue treatment, it is possible to detect transcripts in the fine structures of schistosomes. The protocol is sensitive enough to detect very low abundance targets. This procedure is optimized for tissues derived from S. mansoni adult worms; however, it can be successfully applied to other trematode species.Immunofluorescence allows the detection, visualization, and localization of proteins by using the ability of antibodies to firmly bind to specific antigens. Proteins must be accessible to thorough interaction with the specific antibodies. Different immune evasion mechanisms of parasites are directed to hamper or prevent access of antibodies to critical proteins or virulence factors. The blood fluke Schistosoma mansoni would not survive a day in the host blood capillaries if antibodies were able to readily bind to proteins located at the surface and mediate its attrition and demise by the complement system and/or the FcγR- or FcαR-bearing leukocytes. The worm surface is the area of parasite-host interaction and the route to critical nutrients, but is selectively permeable, allowing access of nutrient molecules but not host antibodies. Gentle procedures, which, however, are not commonly in use in vivo, are required to increase the permeability of the parasite outer membrane shield to just allow access of specific antibodies and identify and localize the proteins at the apical surface. Robust methods involving acetone, methanol, and Triton X-100 treatment lead to disintegration of the dual lipid bilayer cover with exposure of the proteins located in the tegument beneath. Internal proteins may not be accessed except following cryostat or paraffin sectioning. Accordingly, vaccine-induced specific antibodies to the apical surface or tegument proteins are unable to harm intact parasites. Specific antibodies to surface membrane proteins may only add to the action of administered or endo schistosomicides via acceleration of killing and interference with repair of severely and lightly impacted parasites, respectively. Therefore, careful immunofluorescent localization of S. mansoni proteins is important for devising the different control strategies against infection.Schistosomiasis is a chronic neglected tropical disease, highlighted by the presence of Schistosoma worms, which presents in advanced cases in approximately 80 countries, affecting almost 300 million people. The treatment is based on only one drug, praziquantel, a drug discovered in the 1970s that shows moderate efficacy against the adult parasite, but low efficacy against the larval stages of the parasite. Therefore, the use of only one drug has brought concerns and losses on drug-resistance cases, necessitating the development of new effective chemotherapeutic agents against Schistosoma species. One of the strategies that have been implemented in drug development is the computer-aided drug design (CADD), investigating the structural characteristics of the compounds and targets in order to understand their actions and biological activities through 3D virtual manipulation, as the QSAR applied to ligands and molecular docking applied to a respective biological target. These studies help to extract information and characteristics relevant to the activity, as well as to predict potential applications and activity.