The inhibitor Verapamil (Verap), Ethidium Bromide (EtBr) and Calcium Chloride (CaCl2) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Praziquantel was purchased from Merck & Co. (Kenilworth, NJ, USA) and dissolved in Dimethyl Sulfoxide (DMSO) from Sigma-Aldrich, used for stock solutions, which were subsequently diluted to an appropriate concentration in culture media. All solutions were prepared in distilled, sterile water, on the day of the experiments.
Praziquantel
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Sao Tome and Principe, Switzerland
Praziquantel is a pharmaceutical compound primarily used as an antiparasitic agent. It is a crystalline solid and is used in the treatment of various parasitic infections, particularly those caused by trematodes and cestodes. Praziquantel acts by disrupting the calcium homeostasis of the parasites, leading to their death or expulsion from the host.
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Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (7 mentions)
Penicillin, by Merck Group (3 mentions)
Albendazole, by GlaxoSmithKline (2 mentions)
Artesunate, by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Flubendazole, by Merck Group (2 mentions)
Resveratrol resv, by Santa Cruz Biotechnology (2 mentions)
Melatonin mel, by Cayman Chemical (2 mentions)
Auranofin, by Merck Group (2 mentions)
Calcium chloride cacl2, by Merck Group (1 mentions)
Stereomicroscope, by Zeiss (1 mentions)
Serum bovine albumin, by Merck Group (1 mentions)
Ellman s reagent, by Merck Group (1 mentions)
Glutaraldehyde solution, by Merck Group (1 mentions)
Phenol red and l glutamine, by Vitrocell (1 mentions)
Inactivated fetal bovine serum, by Vitrocell (1 mentions)
Penicillin g streptomycin sulfate, by Vitrocell (1 mentions)
M199 medium, by Vitrocell (1 mentions)
Nsaids, by Merck Group (1 mentions)
Methylene blue, by Merck Group (1 mentions)
37 protocols using praziquantel
1
Inhibitor Compound Preparation Protocol
2
Praziquantel for Schistosomiasis Treatment
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The product used was Praziquantel, an existing product manufactured by Merck, SA de CV, Mexico. The drug is also used by the NSCP in MDAs in school-aged children in Senegal. Treatment was administered at the standard dose of 40 mg/kg at the health centre or at the school in each village, in accordance with Senegalese NSCP guidelines and in the presence of community relays, persons which help the health post in the MDAs at the community level. To make sure that the drug was swallowed correctly, each child was asked to open their mouth for verification. If the drugs had not been swallowed, another glass of water was given to the child. For children who vomited the drug, a second full dose was given approximately 30 min after. Children who continued to eliminate schistosome eggs (either viable or not) 1 month after the initial treatment received a second dose of 40 mg/kg.
3
Praziquantel Stock Solution Preparation
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Praziquantel was purchased from Merck & Co. (Kenilworth, NJ, USA) and dissolved in 1% dimethyl sulfoxide (DMSO) from Sigma-Aldrich as described by Melman et al. (2009 (link)), used for stock solution, which was subsequently diluted to appropriate concentrations in culture media.
4
Praziquantel Effectiveness on Liver Enzymes
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Praziquantel was supplied by Merck S.A., Mexico, Mexico. Total bilirubin and alkaline phosphatase kits (Inmesco kits) were purchased from Inmesco, Wied, Germany. Transaminases kits were procured from Quibasa Quimica Basica Ldta, Belo Horizonte, Brasil. The following were products of Sigma-Aldrich, St Louis, USA: Ellman's reagent, serum albumin bovine (SAB), trichloroacetic acid (TCA), thiobarbituric acid (TBA), chemicals used for the preparation of the Biuret reagent and buffers. Methanol and others chemicals were of analytical grade. For parasitological study, we used a stereo-microscope (Zeiss, Hamburg, Germany) and a light microscope (Leica, Wetzlar, Germany). For biochemical analyses, we used the Genesys 20 spectrophotometer (Sigma-Aldrich, St Louis, USA).
5
Deworming and Malaria Treatment Protocol
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Each child was examined by the project nurse and offered praziquantel (40 mg/Kg, Merck KGaA, Darmstadt, Germany) and albendazole (400 mg, GSK, Brentford, UK) irrespective of their infection status, although children observed with a positive malaria RDT were each offered a take-home, 3-day course of artemether–lumefantrine (Coartem® Lonart; Cipla, Mumbai, India) treatment with the first and third treatment administrations overseen by the project nurse. The Ugandan Council for Science and Technology and the Liverpool School of Tropical Medicine granted approvals for this study.
6
Mass Praziquantel Treatment for Schistosomiasis
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Praziquantel was given by Merck KGaA; Darmstadt, Germany to the World Health Organization (WHO) for mass distribution by the national program in the fight against schistosomiasis in Senegal. The Niakhar study area is one of the sites selected for five rounds of annual mass treatment against schistosomiasis based on the prevalence of urinary schistosomiasis higher than 50 % [29 (link)]. This study took place a year before the beginning of mass treatments in the study zone. The drug was made available to us by the national program and was administered at the standard dose of 40 mg/kg at the health centers of the study zone, and in the presence of a nurse and an agent from the national program.
7
Praziquantel Treatment in Infected Pigs
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In two separate experiments infected pigs were treated with a single dose of praziquantel (100 mg/Kg, Merck, Darmstadt, Germany) and sacrificed 48h (n = 4) and 120h later (n = 4). Five untreated infected pigs were used as controls. Two hours before euthanasia, pigs were anesthetized and infused with 2% Evans blue (EB, 80 mg/Kg; Sigma-Aldrich, St. Louis, MO) in normal saline (NaCl 0.85%, Laboratorios Baxter, Colombia, Baxter del Peru) by intravenous injection via the carotid artery. Just after euthanasia, the pigs were perfused with chilled saline solution with heparin (NaCl 0.85% with 10 UI of heparin/mL) and the brain was immediately removed at necropsy.
8
Adult Bioassays for Drug Screening
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All NSAIDs were purchased from Sigma-Aldrich (St. Louis, MO, USA), Cayman Chemical (Ann Arbor, MI, USA) and Toronto Research Chemicals (Toronto, Ontario, Canada). Praziquantel was purchased from Merck (São Paulo, SP, Brazil). Stock solutions for use in adult bioassays in vitro were prepared in Dimethyl sulfoxide (DMSO) at a concentration of 10 mM. All drug tested (n = 73) are in Supplementary Table 1.
Roswell Park Memorial Institute (RPMI 1640) culture medium containing phenol red andl -glutamine, M199 medium, inactivated fetal bovine serum (FBS), penicillin G/streptomycin sulfate, and HEPES buffer were obtained from Vitrocell (Campinas, SP, Brazil). DMSO and glutaraldehyde solution were obtained from Sigma-Aldrich.
Roswell Park Memorial Institute (RPMI 1640) culture medium containing phenol red and
9
Schistosomiasis Screening and Treatment in Ghanaian Pupils
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Study subjects were elementary school pupils assigned by the Ghana Health Service, Ministry of Education and Noguchi Memorial Institute for Medical Research for schistosomiasis examination and treatment. Subjects were recruited if they discharged urine with blood and/ or pain. Informed consent was obtained from Education Authorities and parents and/or guardians of pupils before recruitment. Infected children were treated with 40mg/kg body weight of praziquantel (Merck KGaA, Darmstadt, Germany) as recommended by WHO [12 ]. Approval for this study was given by the Noguchi Memorial Institute for Medical Research Institutional Review Board, referenced NMIMR-IRB CPN 042/06-07 rev 2008.
10
Cluster Randomized Trial for Schistosomiasis
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Ethical approval for the cluster randomized trial was obtained from the Zanzibar Medical Research Ethics Committee (ZAMREC 0003/Sept/011), the "Ethikkomission beiber Basel" (EKBB) in Switzerland (reference no. 236/11), and the Institutional Review Board of the University of Georgia (project no. 2012-10138-0). All aspects of sample collections for the cluster randomized trial were carried out in collaboration with staff of the Neglected Diseases Programme of the Zanzibar Ministry of Health (MoH) in Unguja and the Public Health Laboratory-Ivo de Carneri in Pemba. Shehia and school authorities were informed about the purpose and procedures of the study. Written informed consent was obtained from parents for all minors (children �16 years old) prior to recruitment. All participating adults provided their own signed consent. Participation was voluntary and individuals could withdraw or be withdrawn from the study at any time without obligation. All infected participants were offered treatment with praziquantel (40 mg/kg), donated by Merck KgA and facilitated by the WHO and Zanzibar Ministry of Health, as part of the MDA implemented by the Schistosomiasis Control Initiative (SCI) and MoH.
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