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  1 / 2531 MEDLINE  
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PMID:28792526
Autor:Haserick JR; Klein JA; Costello CE; Samuelson J
Endereço:Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Boston, Massachusetts, United States of America.
Título:Cryptosporidium parvum vaccine candidates are incompletely modified with O-linked-N-acetylgalactosamine or contain N-terminal N-myristate and S-palmitate.
Fonte:PLoS One; 12(8):e0182395, 2017.
ISSN:1932-6203
País de publicação:United States
Idioma:eng
Resumo:Cryptosporidium parvum (studied here) and Cryptosporidium hominis are important causes of diarrhea in infants and immunosuppressed persons. C. parvum vaccine candidates, which are on the surface of sporozoites, include glycoproteins with Ser- and Thr-rich domains (Gp15, Gp40, and Gp900) and a low complexity, acidic protein (Cp23). Here we used mass spectrometry to determine that O-linked GalNAc is present in dense arrays on a glycopeptide with consecutive Ser derived from Gp40 and on glycopeptides with consecutive Thr derived from Gp20, a novel C. parvum glycoprotein with a formula weight of ~20 kDa. In contrast, the occupied Ser or Thr residues in glycopeptides from Gp15 and Gp900 are isolated from one another. Gly at the N-terminus of Cp23 is N-myristoylated, while Cys, the second amino acid, is S-palmitoylated. In summary, C. parvum O-GalNAc transferases, which are homologs of host enzymes, densely modify arrays of Ser or Thr, as well as isolated Ser and Thr residues on C. parvum vaccine candidates. The N-terminus of an immunodominant antigen has lipid modifications similar to those of host cells and other apicomplexan parasites. Mass spectrometric demonstration here of glycopeptides with O-glycans complements previous identification C. parvum O-GalNAc transferases, lectin binding to vaccine candidates, and human and mouse antibodies binding to glycopeptides. The significance of these post-translational modifications is discussed with regards to the function of these proteins and the design of serological tests and vaccines.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Glycoproteins); 0 (Monosaccharides); 0 (Myristates); 0 (Palmitates); 0 (Polysaccharides); 0 (Protozoan Proteins); 0 (Protozoan Vaccines); KM15WK8O5T (Acetylgalactosamine)


  2 / 2531 MEDLINE  
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PMID:28792380
Autor:Perry MD; Corden SA; Lewis White P
Endereço:Public Health Wales Microbiology Cardiff, University Hospital of Wales, Heath Park, Cardiff, UK.
Título:Evaluation of the BD MAX Enteric Parasite Panel for the detection of Cryptosporidium parvum/hominis, Giardia duodenalis and Entamoeba histolytica.
Fonte:J Med Microbiol; 66(8):1118-1123, 2017 Aug.
ISSN:1473-5644
País de publicação:England
Idioma:eng
Resumo:PURPOSE: Conventional laboratory detection methods for gastrointestinal parasites are time consuming, require considerable technical expertise and may suffer from poor analytical sensitivity. This study sought to evaluate the automated BD MAX Enteric Parasite Panel (EPP) for the detection of Cryptosporidium parvum/hominis, Entamoeba histolytica and Giardia duodenalis.Methodolgy. A total of 104 known positive samples (43 Cryptosporidium parvum/hominis and 61 G. duodenalis), 15 simulated samples (E. histolytica and other Entamoeba species) and 745 patient stool samples, submitted for enteric pathogen culture and microscopy, were inoculated into BD MAX EPP sample buffer tubes (SBTs). All specimens were blinded and tested within 7 days of SBT inoculation using the BD MAX EPP assay with results compared to those generated by microscopy.Results/Key findings. Combining the results from the known positive samples and anonymously tested patient samples, the sensitivity of the BD MAX EPP assay was 100 % for both Cryptosporidium spp. and G. duodenalis. Specificities of 99.7 and 98.9 % were calculated for the detection of Cryptosporidium spp. and G. duodenalis respectively. Insufficient clinical specimen data was available to determine the performance of the assay for E. histolytica detection. CONCLUSIONS: The findings of this study indicate that the BD MAX EPP is suitable for the detection of Cryptosporidium parvum/hominis and G. duodenalis from clinical specimens with reduced hands-on time and complexity compared to microscopy. Results for the detection of E. histolytica were promising although further work is required to evaluate the assay for the detection of this pathogen.
Tipo de publicação: EVALUATION STUDIES; JOURNAL ARTICLE
Nome de substância:0 (DNA, Protozoan)


  3 / 2531 MEDLINE  
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PMID:28598342
Autor:Lu P; Amburgey JE; Hill VR; Murphy JL; Schneeberger CL; Arrowood MJ; Yuan T
Endereço:Department of Environmental Science and Spatial Informatics, China University of Mining and Technology, Xuzhou 221116, China E-mail: lupingcumt@126.com; Civil and Environmental Engineering, University of North Carolina at Charlotte, Charlotte, NC 28223, USA.
Título:Removals of cryptosporidium parvum oocysts and cryptosporidium-sized polystyrene microspheres from swimming pool water by diatomaceous earth filtration and perlite-sand filtration.
Fonte:J Water Health; 15(3):374-384, 2017 Jun.
ISSN:1477-8920
País de publicação:England
Idioma:eng
Resumo:Removal of Cryptosporidium-sized microspheres and Cryptosporidium parvum oocysts from swimming pools was investigated using diatomaceous earth (DE) precoat filtration and perlite-sand filtration. In pilot-scale experiments, microsphere removals of up to 2 log were obtained with 0.7 kg·DE/m at a filtration rate of 5 m/h. A slightly higher microsphere removal (2.3 log) was obtained for these DE-precoated filters when the filtration rate was 3.6 m/h. Additionally, pilot-scale perlite-sand filters achieved greater than 2 log removal when at least 0.37 kg/m of perlite was used compared to 0.1-0.4 log removal without perlite both at a surface loading rate of 37 m/h. Full-scale testing achieved 2.7 log of microspheres and oocysts removal when 0.7 kg·DE/m was used at 3.6 m/h. Removals were significantly decreased by a 15-minute interruption of the flow (without any mechanical agitation) to the DE filter in pilot-scale studies, which was not observed in full-scale filters. Microsphere removals were 2.7 log by perlite-sand filtration in a full-scale swimming pool filter operated at 34 m/h with 0.5 kg/m of perlite. The results demonstrate that either a DE precoat filter or a perlite-sand filter can improve the efficiency of removal of microspheres and oocysts from swimming pools over a standard sand filter under the conditions studied.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Polystyrenes); 0SG101ZGK9 (Perlite); 61790-53-2 (Diatomaceous Earth); 7631-86-9 (Silicon Dioxide); LMI26O6933 (Aluminum Oxide)


  4 / 2531 MEDLINE  
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PMID:28541457
Autor:Arnold SLM; Choi R; Hulverson MA; Schaefer DA; Vinayak S; Vidadala RSR; McCloskey MC; Whitman GR; Huang W; Barrett LK; Ojo KK; Fan E; Maly DJ; Riggs MW; Striepen B; Van Voorhis WC
Endereço:Department of Medicine, Division of Allergy and Infectious Disease, Center for Emerging and Reemerging Infectious Disease.
Título:Necessity of Bumped Kinase Inhibitor Gastrointestinal Exposure in Treating Cryptosporidium Infection.
Fonte:J Infect Dis; 216(1):55-63, 2017 Jul 01.
ISSN:1537-6613
País de publicação:United States
Idioma:eng
Resumo:There is a substantial need for novel therapeutics to combat the widespread impact caused by Crytosporidium infection. However, there is a lack of knowledge as to which drug pharmacokinetic (PK) characteristics are key to generate an in vivo response, specifically whether systemic drug exposure is crucial for in vivo efficacy. To identify which PK properties are correlated with in vivo efficacy, we generated physiologically based PK models to simulate systemic and gastrointestinal drug concentrations for a series of bumped kinase inhibitors (BKIs) that have nearly identical in vitro potency against Cryptosporidium but display divergent PK properties. When BKI concentrations were used to predict in vivo efficacy with a neonatal model of Cryptosporidium infection, these concentrations in the large intestine were the sole predictors of the observed in vivo efficacy. The significance of large intestinal BKI exposure for predicting in vivo efficacy was further supported with an adult mouse model of Cryptosporidium infection. This study suggests that drug exposure in the large intestine is essential for generating a superior in vivo response, and that physiologically based PK models can assist in the prioritization of leading preclinical drug candidates for in vivo testing.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Naphthalenes); 0 (Piperidines); 0 (Protein Kinase Inhibitors); 0 (Pyrazoles); 0 (bumped kinase inhibitor 1294)


  5 / 2531 MEDLINE  
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PMID:28520899
Autor:Bolick DT; Mayneris-Perxachs J; Medlock GL; Kolling GL; Papin JA; Swann JR; Guerrant RL
Endereço:Division of Infectious Diseases and International Health, UVA Center for Global Health, University of Virginia, Charlottesville.
Título:Increased Urinary Trimethylamine N-Oxide Following Cryptosporidium Infection and Protein Malnutrition Independent of Microbiome Effects.
Fonte:J Infect Dis; 216(1):64-71, 2017 Jul 01.
ISSN:1537-6613
País de publicação:United States
Idioma:eng
Resumo:Cryptosporidium infections have been associated with growth stunting, even in the absence of diarrhea. Having previously detailed the effects of protein deficiency on both microbiome and metabolome in this model, we now describe the specific gut microbial and biochemical effects of Cryptosporidium infection. Protein-deficient mice were infected with Cryptosporidium parvum oocysts for 6-13 days and compared with uninfected controls. Following infection, there was an increase in the urinary excretion of choline- and amino-acid-derived metabolites. Conversely, infection reduced the excretion of the microbial-host cometabolite (3-hydroxyphenyl)propionate-sulfate and disrupted metabolites involved in the tricarboxylic acid (TCA) cycle. Correlation analysis of microbial and biochemical profiles resulted in associations between various microbiota members and TCA cycle metabolites, as well as some microbial-specific degradation products. However, no correlation was observed between the majority of the infection-associated metabolites and the fecal bacteria, suggesting that these biochemical perturbations are independent of concurrent changes in the relative abundance of members of the microbiota. We conclude that cryptosporidial infection in protein-deficient mice can mimic some metabolic changes seen in malnourished children and may help elucidate our understanding of long-term metabolic consequences of early childhood enteric infections.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Biomarkers); 0 (Lipocalin-2); 0 (Methylamines); 126469-30-5 (Lcn2 protein, mouse); EC 1.11.1.7 (Peroxidase); FLD0K1SJ1A (trimethyloxamine)


  6 / 2531 MEDLINE  
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PMID:28506035
Autor:Sim S; Won J; Kim JW; Kim K; Park WY; Yu JR
Endereço:Department of Environmental and Tropical Medicine & International Healthcare Research Institute, Konkuk University School of Medicine, Seoul 05029, Korea.
Título:Simultaneous Molecular Detection of and from Raw Vegetables in Korea.
Fonte:Korean J Parasitol; 55(2):137-142, 2017 Apr.
ISSN:1738-0006
País de publicação:Korea (South)
Idioma:eng
Resumo:and are well-known coccidian protozoa that can cause waterborne and foodborne diarrheal illnesses. There have been a few reports regarding contamination in different vegetables with , but no data are available regarding the sources of infections in Korea. In the present study, we collected 6 kinds of vegetables (perilla leaves, winter-grown cabbages, chives, sprouts, blueberries, and cherry tomatoes) from July 2014 to June 2015, and investigated contamination by these 2 protozoa using multiplex quantitative real-time PCR. Among 404 vegetables, and were detected in 31 (7.7%) and 5 (1.2%) samples, respectively. In addition, was isolated from all 6 kinds of vegetables, whereas was detected in 4 kinds of vegetables (except perilla leaves and chives). (17.8%) and (2.9%) had the highest detection rates in chives and winter-grown cabbages, respectively. was detected all year long; however, was detected only from October to January. In 2 samples (sprout and blueberry), both and were detected. Further investigations using I restriction enzyme fragmentation and nested PCR confirmed and , respectively. In conclusion, we detected in vegetables for the first time in Korea. This suggests that screening should be employed to prevent these protozoal infections in Korea.
Tipo de publicação: JOURNAL ARTICLE


  7 / 2531 MEDLINE  
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PMID:28445027
Autor:Varga MJ; Dzierlenga MW; Schwartz SD
Endereço:Department of Chemistry and Biochemistry, University of Arizona , 1306 East University Boulevard, Tucson, Arizona 85721, United States.
Título:Structurally Linked Dynamics in Lactate Dehydrogenases of Evolutionarily Distinct Species.
Fonte:Biochemistry; 56(19):2488-2496, 2017 May 16.
ISSN:1520-4995
País de publicação:United States
Idioma:eng
Resumo:We present new findings about how primary and secondary structure affects the role of fast protein motions in the reaction coordinates of enzymatic reactions. Using transition path sampling and committor distribution analysis, we examined the difference in the role of these fast protein motions in the reaction coordinate of lactate dehydrogenases (LDHs) of Apicomplexa organisms Plasmodium falciparum and Cryptosporidium parvum. Having evolved separately from a common malate dehydrogenase ancestor, the two enzymes exhibit several important structural differences, notably a five-amino acid insertion in the active site loop of P. falciparum LDH. We find that these active site differences between the two organisms' LDHs likely cause a decrease in the contribution of the previously determined LDH rate-promoting vibration to the reaction coordinate of P. falciparum LDH compared to that of C. parvum LDH, specifically in the coupling of the rate-promoting vibration and the hydride transfer. This effect, while subtle, directly shows how changes in structure near the active site of LDH alter catalytically important motions. Insights provided by studying these alterations would prove to be useful in identifying LDH inhibitors that specifically target the isozymes of these parasitic organisms.
Tipo de publicação: COMPARATIVE STUDY; JOURNAL ARTICLE
Nome de substância:0 (Isoenzymes); 0 (Protozoan Proteins); EC 1.1.- (Lactate Dehydrogenases)


  8 / 2531 MEDLINE  
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PMID:28329187
Autor:Hulverson MA; Vinayak S; Choi R; Schaefer DA; Castellanos-Gonzalez A; Vidadala RSR; Brooks CF; Herbert GT; Betzer DP; Whitman GR; Sparks HN; Arnold SLM; Rivas KL; Barrett LK; White AC; Maly DJ; Riggs MW; Striepen B; Van Voorhis WC; Ojo KK
Endereço:Division of Allergy & Infectious Disease, Center for Emerging & Reemerging Infectious Disease (CERID), University of Washington, Seattle, Washington 98109, USA.
Título:Bumped-Kinase Inhibitors for Cryptosporidiosis Therapy.
Fonte:J Infect Dis; 215(8):1275-1284, 2017 04 15.
ISSN:1537-6613
País de publicação:United States
Idioma:eng
Resumo:Bumped kinase inhibitors (BKIs) of Cryptosporidium parvum calcium-dependent protein kinase 1 (CpCDPK1) are leading candidates for treatment of cryptosporidiosis-associated diarrhea. Potential cardiotoxicity related to anti-human ether-à-go-go potassium channel (hERG) activity of the first-generation anti-Cryptosporidium BKIs triggered further testing for efficacy. A luminescence assay adapted for high-throughput screening was used to measure inhibitory activities of BKIs against C. parvum in vitro. Furthermore, neonatal and interferon γ knockout mouse models of C. parvum infection identified BKIs with in vivo activity. Additional iterative experiments for optimum dosing and selecting BKIs with minimum levels of hERG activity and frequencies of other safety liabilities included those that investigated mammalian cell cytotoxicity, C. parvum proliferation inhibition in vitro, anti-human Src inhibition, hERG activity, in vivo pharmacokinetic data, and efficacy in other mouse models. Findings of this study suggest that fecal concentrations greater than parasite inhibitory concentrations correlate best with effective therapy in the mouse model of cryptosporidiosis, but a more refined model for efficacy is needed.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Antiprotozoal Agents); 0 (Protein Kinase Inhibitors)


  9 / 2531 MEDLINE  
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PMID:28192123
Autor:Feng Y; Li N; Roellig DM; Kelley A; Liu G; Amer S; Tang K; Zhang L; Xiao L
Endereço:College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China; State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China. Electronic address: yyfeng@scau.edu.cn.
Título:Comparative genomic analysis of the IId subtype family of Cryptosporidium parvum.
Fonte:Int J Parasitol; 47(5):281-290, 2017 Apr.
ISSN:1879-0135
País de publicação:England
Idioma:eng
Resumo:Host adaptation is known to occur in Cryptosporidium parvum, with IIa and IId subtype families preferentially infecting calves and lambs, respectively. To improve our understanding of the genetic basis of host adaptation in Cryptosporidium parvum, we sequenced the genomes of two IId specimens and one IIa specimen from China and Egypt using the Illumina technique and compared them with the published IIa IOWA genome. Sequence data were obtained for >99.3% of the expected genome. Comparative genomic analysis identified differences in numbers of three subtelomeric gene families between sequenced genomes and the reference genome, including those encoding SKSR secretory proteins, the MEDLE family of secretory proteins, and insulinase-like proteases. These gene gains and losses compared with the reference genome were confirmed by PCR analysis. Altogether, 5,191-5,766 single nucleotide variants were seen between genomes sequenced in this study and the reference genome, with most SNVs occurring in subtelomeric regions of chromosomes 1, 4, and 6. The most highly polymorphic genes between IIa and IId encode mainly invasion-associated and immunodominant mucin proteins, and other families of secretory proteins. Further studies are needed to verify the biological significance of these genomic differences.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (DNA Transposable Elements); 0 (DNA, Protozoan)


  10 / 2531 MEDLINE  
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PMID:28179475
Autor:Haserick JR; Leon DR; Samuelson J; Costello CE
Endereço:From the ‡Center for Biomedical Mass Spectrometry, Department of Biochemistry, Cell Biology and Genomics, Boston University School of Medicine, Boston, Massachusetts 02118 and.
Título:Asparagine-Linked Glycans of Contain a Single Long Arm, Are Barely Processed in the Endoplasmic Reticulum (ER) or Golgi, and Show a Strong Bias for Sites with Threonine.
Fonte:Mol Cell Proteomics; 16(4 suppl 1):S42-S53, 2017 Apr.
ISSN:1535-9484
País de publicação:United States
Idioma:eng
Resumo:causes severe diarrhea in infants in developing countries and in immunosuppressed persons, including those with AIDS. We are interested in the Asn-linked glycans ( -glycans) of , because (1) the -glycan precursor is predicted to contain five mannose and two glucose residues on a single long arm nine mannose and three glucose residues on the three-armed structure common in host -glycans, (2) is a rare eukaryote that lacks the machinery for -glycan-dependent quality control of protein folding in the lumen of the Endoplasmic Reticulum (ER), and (3) ER and Golgi mannosidases, as well as glycosyltransferases that build complex -glycans, are absent from the predicted proteome. The -glycans reported here, which were determined using a combination of collision-induced dissociation and electronic excitation dissociation, contain a single, unprocessed mannose arm ± terminal glucose on the trimannosyl chitobiose core. Upon nanoUPLC-MS/MS separation and analysis of the tryptic peptides, the total ion and extracted oxonium ion chromatograms delineated 32 peptides with occupied -glycan sites; these were derived from 16 glycoproteins. Although the number of potential -glycan sites with Thr (NxT) is only about twice that with Ser (NxS), almost 90% of the occupied -glycan sites contain NxT. The two most abundant proteins modified with -glycans were an immunodominant antigen on the surface of sporozoites (gp900) and the possible oocyst wall protein 1 (POWP1). Seven other glycoproteins with -glycans were unique to ; five shared common ancestry with other apicomplexans; two glycoproteins shared common ancestry with many organisms. In summary, -glycans are remarkable for the absence of ER and Golgi modification and for the strong bias toward occupancy of -glycan motifs containing Thr.
Tipo de publicação: JOURNAL ARTICLE
Nome de substância:0 (Glycoproteins); 0 (Polysaccharides); 0 (Protozoan Proteins); 452VLY9402 (Serine); 7006-34-0 (Asparagine)



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