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  1 / 4025 MEDLINE  
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[PMID]:29250543
[Au] Autor:Li Z; Su L; Duan X; Wu D; Wu J
[Ad] Endereço:State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
[Ti] Título:Efficient Expression of Maltohexaose-Forming -Amylase from in SP3 and Its Use in Maltose Production.
[So] Source:Biomed Res Int;2017:5479762, 2017.
[Is] ISSN:2314-6141
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The maltohexaose-forming, Ca -independent -amylase gene from (AmyMH) was efficiently expressed in SP3. To improve the production of AmyMH in SP3, the temperature and initial pH of culture medium were optimized. In addition, single-factor and response surface methodologies were pursued to optimize culture medium. Addition of proline to the culture medium significantly improved the production of recombinant -amylase in SP3. This improvement may result from improved cellular integrity of recombinant SP3 in existence of proline. Culture medium optimization resulted in an 8-fold improvement in -amylase yield, which reached 1.72 × 10 U·mL . The recombinant -amylase was applied to the production of maltose on a laboratory scale. A maltose content of 90.72%, which could be classified as an extremely high maltose syrup, could be achieved using 15% (m/v) corn starch as the substrate. This study demonstrated that the SP3 expression system was able to produce substantial quantities of recombinant -amylase that has potential application in the starch industry.
[Mh] Termos MeSH primário: Brevibacillus/metabolismo
Geobacillus stearothermophilus/enzimologia
Maltose/metabolismo
Oligossacarídeos/metabolismo
alfa-Amilases/metabolismo
[Mh] Termos MeSH secundário: Proteínas de Bactérias/genética
Proteínas de Bactérias/metabolismo
Reatores Biológicos
Brevibacillus/genética
Meios de Cultura
Fermentação
Geobacillus stearothermophilus/genética
Glucose/metabolismo
Maltose/análise
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
alfa-Amilases/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Culture Media); 0 (Oligosaccharides); 0 (Recombinant Proteins); 34620-77-4 (maltohexaose); 69-79-4 (Maltose); EC 3.2.1.1 (alpha-Amylases); IY9XDZ35W2 (Glucose)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180129
[Lr] Data última revisão:
180129
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171219
[St] Status:MEDLINE
[do] DOI:10.1155/2017/5479762


  2 / 4025 MEDLINE  
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[PMID]:28547803
[Au] Autor:He Y; Zhang H; Wen N; Hu R; Wu G; Zeng Y; Li X; Miao X
[Ad] Endereço:College of Food Science and Technology, Hainan University, Haikou, China.
[Ti] Título:Effects of maltose and lysine treatment on coffee aroma by flash gas chromatography electronic nose and gas chromatography-mass spectrometry.
[So] Source:J Sci Food Agric;98(1):154-165, 2018 Jan.
[Is] ISSN:1097-0010
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Arabica coffee is a sub-tropical agricultural product in China. Coffee undergoes a series of thermal reactions to form abundant volatile profiles after roasting, so it loses a lot of reducing sugars and amino acids. Adding carbonyl compounds with amino acids before roasting could ensure the nutrition and flavour of coffee. The technology is versatile for the development of coffee roasting process. This investigation evaluates the effects of combining maltose and lysine (Lys) to modify coffee aroma and the possibly related mechanisms. Arabica coffee was pretreated with a series of solvent ratios of maltose and Lys with an identical concentration (0.25 mol L ) before microwave heating. RESULTS: It was found that the combination of maltose and Lys significantly (P ≤ 0.05) influenced quality indices of coffee (pH and browning degree). Ninety-six aromatic volatiles have been isolated and identified. Twelve volatile profiles revealed the relationship between fragrance difference and compound content in coffee. Moreover, coffee aroma was modified by a large number of volatiles with different chemical classes and character. CONCLUSION: Thus, our results suggest that the combination of reagents changed overall aroma quality through a series of complex thermal reactions, especially the ratio of Lys/maltose over 2:1. © 2017 Society of Chemical Industry.
[Mh] Termos MeSH primário: Coffea/química
Café/química
Aditivos Alimentares/análise
Manipulação de Alimentos/métodos
Lisina/análise
Maltose/análise
Compostos Orgânicos Voláteis/química
[Mh] Termos MeSH secundário: Culinária
Nariz Eletrônico
Cromatografia Gasosa-Espectrometria de Massas
Temperatura Alta
Odorantes/análise
Sementes/química
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Coffee); 0 (Food Additives); 0 (Volatile Organic Compounds); 69-79-4 (Maltose); K3Z4F929H6 (Lysine)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180102
[Lr] Data última revisão:
180102
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170527
[St] Status:MEDLINE
[do] DOI:10.1002/jsfa.8450


  3 / 4025 MEDLINE  
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[PMID]:28455338
[Au] Autor:Joyet P; Mokhtari A; Riboulet-Bisson E; Blancato VS; Espariz M; Magni C; Hartke A; Deutscher J; Sauvageot N
[Ad] Endereço:Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, Jouy-en-Josas, France.
[Ti] Título:Enzymes Required for Maltodextrin Catabolism in Enterococcus faecalis Exhibit Novel Activities.
[So] Source:Appl Environ Microbiol;83(13), 2017 Jul 01.
[Is] ISSN:1098-5336
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Maltose and maltodextrins are formed during the degradation of starch or glycogen. Maltodextrins are composed of a mixture of maltooligosaccharides formed by α-1,4- but also some α-1,6-linked glucosyl residues. The α-1,6-linked glucosyl residues are derived from branching points in the polysaccharides. In , maltotriose is mainly transported and phosphorylated by a phosphoenolpyruvate:carbohydrate phosphotransferase system. The formed maltotriose-6″-phosphate is intracellularly dephosphorylated by a specific phosphatase, MapP. In contrast, maltotetraose and longer maltooligosaccharides up to maltoheptaose are taken up without phosphorylation via the ATP binding cassette transporter MdxEFG-MsmX. We show that the maltose-producing maltodextrin hydrolase MmdH (GenBank accession no. EFT41964) in strain JH2-2 catalyzes the first catabolic step of α-1,4-linked maltooligosaccharides. The purified enzyme converts even-numbered α-1,4-linked maltooligosaccharides (maltotetraose, etc.) into maltose and odd-numbered (maltotriose, etc.) into maltose and glucose. Inactivation of therefore prevents the growth of on maltooligosaccharides ranging from maltotriose to maltoheptaose. Surprisingly, MmdH also functions as a maltogenic α-1,6-glucosidase, because it converts the maltotriose isomer isopanose into maltose and glucose. In addition, contains a glucose-producing α-1,6-specific maltodextrin hydrolase (GenBank accession no. EFT41963, renamed GmdH). This enzyme converts panose, another maltotriose isomer, into glucose and maltose. A mutant had therefore lost the capacity to grow on panose. The genes and are organized in an operon together with GenBank accession no. (renamed ). Purified MmgT transfers glucosyl residues from one α-1,4-linked maltooligosaccharide molecule to another. For example, it catalyzes the disproportionation of maltotriose by transferring a glucosyl residue to another maltotriose molecule, thereby forming maltotetraose and maltose together with a small amount of maltopentaose. The utilization of maltodextrins by has been shown to increase the virulence of this nosocomial pathogen. However, little is known about how this organism catabolizes maltodextrins. We identified two enzymes involved in the metabolism of various α-1,4- and α-1,6-linked maltooligosaccharides. We found that one of them functions as a maltose-producing α-glucosidase with relaxed linkage specificity (α-1,4 and α-1,6) and exo- and endoglucosidase activities. A third enzyme, which resembles amylomaltase, exclusively transfers glucosyl residues from one maltooligosaccharide molecule to another. Similar enzymes are present in numerous other , such as streptococci and lactobacilli, suggesting that these organisms follow the same maltose degradation pathway as .
[Mh] Termos MeSH primário: Proteínas de Bactérias/metabolismo
Enterococcus faecalis/enzimologia
Hidrolases/metabolismo
Polissacarídeos/biossíntese
[Mh] Termos MeSH secundário: Transportadores de Cassetes de Ligação de ATP/genética
Transportadores de Cassetes de Ligação de ATP/metabolismo
Proteínas de Bactérias/genética
Enterococcus faecalis/genética
Enterococcus faecalis/metabolismo
Hidrolases/genética
Maltose/metabolismo
Oligossacarídeos/metabolismo
Óperon
Trissacarídeos/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (ATP-Binding Cassette Transporters); 0 (Bacterial Proteins); 0 (Oligosaccharides); 0 (Polysaccharides); 0 (Trisaccharides); 0 (maltooligosaccharides); 639K0T34IK (maltotriose); 69-79-4 (Maltose); 7CVR7L4A2D (maltodextrin); EC 3.- (Hydrolases)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171225
[Lr] Data última revisão:
171225
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170430
[St] Status:MEDLINE


  4 / 4025 MEDLINE  
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[PMID]:28941825
[Au] Autor:Smith AT; Sestok AE
[Ad] Endereço:Department of Chemistry and Biochemistry, University of Maryland, Baltimore County, Baltimore, MD, 21250, USA. Electronic address: smitha@umbc.edu.
[Ti] Título:Expression and purification of functionally active ferrous iron transporter FeoB from Klebsiella pneumoniae.
[So] Source:Protein Expr Purif;142:1-7, 2018 Feb.
[Is] ISSN:1096-0279
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The acquisition of ferrous iron (Fe ) is an important virulence factor utilized by several hospital-acquired (nosocomial) pathogens such as Klebsiella pneumoniae to establish infection within human hosts. Virtually all bacteria use the ferrous iron transport system (Feo) to acquire ferrous iron from their environments, which are often biological niches that stabilize Fe relative to Fe . However, the details of this process remain poorly understood, likely owing to the few expression and purification systems capable of supplying sufficient quantities of the chief component of the Feo system, the integral membrane GTPase FeoB. This bottleneck has undoubtedly hampered efforts to understand this system in order to target it for therapeutic intervention. In this study, we describe the expression, solubilization, and purification of the Fe transporter from K. pneumoniae, KpFeoB. We show that this protein may be heterologously overexpressed in Escherichia coli as the host organism. After testing several different commercially-available detergents, we have developed a solubilization and purification protocol that produces milligram quantities of KpFeoB with sufficient purity for enzymatic and biophysical analyses. Importantly, we demonstrate that KpFeoB displays robust GTP hydrolysis activity (k of ∼10 s ) in the absence of any additional stimulatory factors. Our findings suggest that K. pneumoniae may be capable of using its Feo system to drive Fe import in an active manner.
[Mh] Termos MeSH primário: Proteínas de Bactérias/genética
Proteínas de Transporte de Cátions/genética
Guanosina Trifosfato/metabolismo
Ferro/metabolismo
Klebsiella pneumoniae/química
[Mh] Termos MeSH secundário: Proteínas de Bactérias/química
Proteínas de Bactérias/isolamento & purificação
Proteínas de Bactérias/metabolismo
Proteínas de Transporte de Cátions/química
Proteínas de Transporte de Cátions/isolamento & purificação
Proteínas de Transporte de Cátions/metabolismo
Cátions Bivalentes
Clonagem Molecular
Detergentes/química
Ensaios Enzimáticos
Escherichia coli/genética
Escherichia coli/metabolismo
Expressão Gênica
Hidrólise
Transporte de Íons
Cinética
Klebsiella pneumoniae/enzimologia
Maltose/análogos & derivados
Maltose/química
Plasmídeos/química
Plasmídeos/metabolismo
Polietilenoglicóis/química
Conformação Proteica em alfa-Hélice
Proteínas Recombinantes/química
Proteínas Recombinantes/genética
Proteínas Recombinantes/isolamento & purificação
Proteínas Recombinantes/metabolismo
Solubilidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Cation Transport Proteins); 0 (Cations, Divalent); 0 (Detergents); 0 (Recombinant Proteins); 0 (dodecyl maltopyranoside); 3055-98-9 (dodecyloctaethyleneglycol monoether); 30IQX730WE (Polyethylene Glycols); 69-79-4 (Maltose); 86-01-1 (Guanosine Triphosphate); E1UOL152H7 (Iron)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171128
[Lr] Data última revisão:
171128
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170925
[St] Status:MEDLINE


  5 / 4025 MEDLINE  
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[PMID]:28973244
[Au] Autor:Kim YW; Yang H
[Ad] Endereço:Graduate School of Cancer Science and Policy, National Cancer Center, Goyang, Republic of Korea.
[Ti] Título:Ferric Carboxymaltose to Treat Isovolemic Anemia-Reply.
[So] Source:JAMA;318(13):1281-1282, 2017 10 03.
[Is] ISSN:1538-3598
[Cp] País de publicação:United States
[La] Idioma:eng
[Mh] Termos MeSH primário: Compostos Férricos
Maltose
[Mh] Termos MeSH secundário: Anemia
Anemia Ferropriva
Seres Humanos
Maltose/análogos & derivados
[Pt] Tipo de publicação:LETTER; COMMENT
[Nm] Nome de substância:
0 (Ferric Compounds); 6897GXD6OE (ferric carboxymaltose); 69-79-4 (Maltose)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:171004
[St] Status:MEDLINE
[do] DOI:10.1001/jama.2017.11651


  6 / 4025 MEDLINE  
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[PMID]:28973241
[Au] Autor:Yu Z; Li T
[Ad] Endereço:Department of Neurosurgery, Affiliated Hospital of Chengdu University, Chengdu, China.
[Ti] Título:Ferric Carboxymaltose to Treat Isovolemic Anemia.
[So] Source:JAMA;318(13):1281, 2017 10 03.
[Is] ISSN:1538-3598
[Cp] País de publicação:United States
[La] Idioma:eng
[Mh] Termos MeSH primário: Compostos Férricos
Maltose
[Mh] Termos MeSH secundário: Anemia
Anemia Ferropriva
Seres Humanos
Maltose/análogos & derivados
[Pt] Tipo de publicação:LETTER; COMMENT
[Nm] Nome de substância:
0 (Ferric Compounds); 6897GXD6OE (ferric carboxymaltose); 69-79-4 (Maltose)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:171004
[St] Status:MEDLINE
[do] DOI:10.1001/jama.2017.11637


  7 / 4025 MEDLINE  
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[PMID]:28850567
[Au] Autor:Pullicin AJ; Penner MH; Lim J
[Ad] Endereço:Department of Food Science & Technology, Oregon State University, Corvallis, Oregon, United States of America.
[Ti] Título:Human taste detection of glucose oligomers with low degree of polymerization.
[So] Source:PLoS One;12(8):e0183008, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Studies have reported that some animals, including humans, can taste mixtures of glucose oligomers (i.e., maltooligosaccharides, MOS) and that their detection is independent of the known T1R2/T1R3 sweet taste receptor. In an effort to understand potential mechanisms underlying the taste perception of glucose oligomers in humans, this study was designed to investigate: 1) the variability of taste sensitivity to MOS with low degree-of-polymerization (DP), and 2) the potential role of hT1R2/T1R3 in the MOS taste detection. To address these objectives, a series of food grade, narrow-DP-range MOS were first prepared (DP 3, 3-4, 5-6, and 6-7) by fractionating disperse saccharide mixtures. Subjects were then asked to discriminate these MOS stimuli as well as glucose (DP 1) and maltose (DP 2) from blanks after the stimuli were swabbed on the tongue. All stimuli were presented at 75 mM with and without a sweet taste inhibitor (lactisole). An α-glucosidase inhibitor (acarbose) was added to all test stimuli to prevent oral digestion of glucose oligomers. Results showed that all six stimuli were detected with similar discriminability in normal tasting conditions. When the sweet receptor was inhibited, DP 1, 2, and 3 were not discriminated from blanks. In contrast, three higher-DP paired MOS stimuli (DP 3-4, 5-6, and 6-7) were discriminated from blanks at a similar degree. Overall, these results support the presence of a sweet-independent taste perception mechanism that is stimulated by MOS greater than three units.
[Mh] Termos MeSH primário: Discriminação (Psicologia)/efeitos dos fármacos
Oligossacarídeos/administração & dosagem
Edulcorantes/administração & dosagem
Percepção Gustatória/efeitos dos fármacos
Paladar/efeitos dos fármacos
[Mh] Termos MeSH secundário: Adulto
Derivados de Benzeno/farmacologia
Feminino
Glucose/administração & dosagem
Seres Humanos
Masculino
Maltose/administração & dosagem
Meia-Idade
Paladar/fisiologia
Papilas Gustativas/efeitos dos fármacos
Percepção Gustatória/fisiologia
Adulto Jovem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Benzene Derivatives); 0 (Oligosaccharides); 0 (Sweetening Agents); 0 (maltooligosaccharides); 69-79-4 (Maltose); IY9XDZ35W2 (Glucose); ZU3D90W5GZ (lactisole)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171019
[Lr] Data última revisão:
171019
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170830
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0183008


  8 / 4025 MEDLINE  
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[PMID]:28701470
[Au] Autor:van Veldhuisen DJ; Ponikowski P; van der Meer P; Metra M; Böhm M; Doletsky A; Voors AA; Macdougall IC; Anker SD; Roubert B; Zakin L; Cohen-Solal A; EFFECT-HF Investigators
[Ad] Endereço:From Department of Cardiology, University Medical Center Groningen, University of Groningen, The Netherlands (D.J.v.V., P.v.d.M., A.A.V.); Department of Heart Diseases, Medical University, Clinical Military Hospital, Wroclaw, Poland (P.P.); Department of Cardiology, University Hospital, Brescia, Ita
[Ti] Título:Effect of Ferric Carboxymaltose on Exercise Capacity in Patients With Chronic Heart Failure and Iron Deficiency.
[So] Source:Circulation;136(15):1374-1383, 2017 Oct 10.
[Is] ISSN:1524-4539
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Iron deficiency is common in patients with heart failure (HF) and is associated with reduced exercise capacity and poor outcomes. Whether correction of iron deficiency with (intravenous) ferric carboxymaltose (FCM) affects peak oxygen consumption [peak VO ], an objective measure of exercise intolerance in HF, has not been examined. METHODS: We studied patients with systolic HF (left ventricular ejection fraction ≤45%) and mild to moderate symptoms despite optimal HF medication. Patients were randomized 1:1 to treatment with FCM for 24 weeks or standard of care. The primary end point was the change in peak VO from baseline to 24 weeks. Secondary end points included the effect on hematinic and cardiac biomarkers, quality of life, and safety. For the primary analysis, patients who died had a value of 0 imputed for 24-week peak VO . Additional sensitivity analyses were performed to determine the impact of imputation of missing peak VO data. RESULTS: A total of 172 patients with HF were studied and received FCM (n=86) or standard of care (control group, n=86). At baseline, the groups were well matched; mean age was 64 years, 75% were male, mean left ventricular ejection fraction was 32%, and peak VO was 13.5 mL/min/kg. FCM significantly increased serum ferritin and transferrin saturation. At 24 weeks, peak VO had decreased in the control group (least square means -1.19±0.389 mL/min/kg) but was maintained on FCM (-0.16±0.387 mL/min/kg; =0.020 between groups). In a sensitivity analysis, in which missing data were not imputed, peak VO at 24 weeks decreased by -0.63±0.375 mL/min/kg in the control group and by -0.16±0.373 mL/min/kg in the FCM group; =0.23 between groups). Patients' global assessment and functional class as assessed by the New York Heart Association improved on FCM versus standard of care. CONCLUSIONS: Treatment with intravenous FCM in patients with HF and iron deficiency improves iron stores. Although a favorable effect on peak VO was observed on FCM, compared with standard of care in the primary analysis, this effect was highly sensitive to the imputation strategy for peak VO among patients who died. Whether FCM is associated with an improved outcome in these high-risk patients needs further study. CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier: NCT01394562.
[Mh] Termos MeSH primário: Tolerância ao Exercício/efeitos dos fármacos
Compostos Férricos/administração & dosagem
Insuficiência Cardíaca
Maltose/análogos & derivados
[Mh] Termos MeSH secundário: Idoso
Biomarcadores/sangue
Feminino
Ferritinas/sangue
Insuficiência Cardíaca/sangue
Insuficiência Cardíaca/tratamento farmacológico
Insuficiência Cardíaca/fisiopatologia
Seres Humanos
Ferro/deficiência
Masculino
Maltose/administração & dosagem
Meia-Idade
Oxigênio/sangue
Qualidade de Vida
Volume Sistólico/efeitos dos fármacos
Transferrina/metabolismo
[Pt] Tipo de publicação:CLINICAL TRIAL; JOURNAL ARTICLE; MULTICENTER STUDY
[Nm] Nome de substância:
0 (Biomarkers); 0 (Ferric Compounds); 0 (Transferrin); 6897GXD6OE (ferric carboxymaltose); 69-79-4 (Maltose); 9007-73-2 (Ferritins); E1UOL152H7 (Iron); S88TT14065 (Oxygen)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:170714
[St] Status:MEDLINE
[do] DOI:10.1161/CIRCULATIONAHA.117.027497


  9 / 4025 MEDLINE  
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[PMID]:28701129
[Au] Autor:Low MS; Grigoriadis G
[Ad] Endereço:Monash Health, Melbourne, VIC george.grigoriadis@monash.edu.
[Ti] Título:Iron deficiency and new insights into therapy.
[So] Source:Med J Aust;207(2):81-87, 2017 Jul 17.
[Is] ISSN:1326-5377
[Cp] País de publicação:Australia
[La] Idioma:eng
[Ab] Resumo:Iron deficiency and iron deficiency anaemia remain prevalent in Australia. The groups at highest risk are pre-menopausal women, socially disadvantaged people and those of Indigenous background. Diagnosing iron deficiency using a full blood examination and iron studies can be difficult and can be further complicated by concomitant inflammation. Results of iron studies should always be interpreted as an overall picture rather than focusing on individual parameters. In difficult clinical scenarios, soluble transferrin receptor assays can be useful. Management of iron deficiency involves identification and treatment of the cause of iron deficiency, as well as effective iron replacement. Clinicians should always take a detailed history and perform a comprehensive physical examination of a patient with iron deficiency. Patients should be monitored even if a likely cause of iron deficiency is identified. Patients who fail to respond to iron replacement or maintain iron status should be referred for further investigation, including endoscopy to exclude internal bleeding. Both enteral and parenteral iron are effective at replacing iron. For most adult patients, we recommend trialling daily oral iron (30-100 mg of elemental iron) as the first-line therapy. Safety and efficacy of intravenous iron infusions have improved with the availability of a newer formulation, ferric carboxymaltose. Patients who fail to respond to oral iron replacement can be safely managed with intravenous iron. Blood transfusion for iron deficiency anaemia should be reserved for life-threatening situations and should always be followed by appropriate iron replacement.
[Mh] Termos MeSH primário: Anemia Ferropriva/epidemiologia
Anemia Ferropriva/terapia
Compostos Férricos/administração & dosagem
Ferro/sangue
Maltose/análogos & derivados
[Mh] Termos MeSH secundário: Administração Oral
Anemia Ferropriva/etiologia
Austrália/epidemiologia
Transfusão de Sangue
Medula Óssea/patologia
Criança
Feminino
Compostos Férricos/efeitos adversos
Seres Humanos
Infusões Intravenosas
Ferro/administração & dosagem
Ferro/deficiência
Maltose/administração & dosagem
Maltose/efeitos adversos
Gravidez
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Ferric Compounds); 6897GXD6OE (ferric carboxymaltose); 69-79-4 (Maltose); E1UOL152H7 (Iron)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170714
[St] Status:MEDLINE


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[PMID]:28692650
[Au] Autor:Abeyrathne PD; Grigorieff N
[Ad] Endereço:Howard Hughes Medical Institute, Janelia Research Campus, Helix Drive, Ashburn, VA, United States of America.
[Ti] Título:Expression, purification, and contaminant detection for structural studies of Ralstonia metallidurance ClC protein rm1.
[So] Source:PLoS One;12(7):e0180163, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Single-particle electron cryo-microscopy (cryo-EM) has become a popular method for high-resolution study of the structural and functional properties of proteins. However, sufficient expression and purification of membrane proteins holds many challenges. We describe methods to overcome these obstacles using ClC-rm1, a prokaryotic chloride channel (ClC) family protein from Ralstonia metallidurans, overexpressed in Escherichia coli (E. coli) BL21(DE3) strain. Mass spectrometry and electron microscopy analyses of purified samples revealed multiple contaminants that can obfuscate results of subsequent high-resolution structural analysis. Here we describe the systematic optimization of sample preparation procedures, including expression systems, solubilization techniques, purification protocols, and contamination detection. We found that expressing ClC-rm1 in E. coli BL21(DE3) and using n-dodecyl-ß-D-maltopyranoside as a detergent for solubilization and purification steps resulted in the highest quality samples of those we tested. However, although protein yield, sample stability, and the resolution of structural detail were improved following these changes, we still detected contaminants including Acriflavine resistant protein AcrB. AcrB was particularly difficult to remove as it co-purified with ClC-rm1 due to four intrinsic histidine residues at its C-terminus that bind to affinity resins. We were able to obtain properly folded pure ClC-rm1 by adding eGFP to the C-terminus and overexpressing the protein in the ΔacrB variant of the JW0451-2 E. coli strain.
[Mh] Termos MeSH primário: Proteínas de Bactérias/química
Proteínas de Bactérias/isolamento & purificação
Canais de Cloreto/química
Canais de Cloreto/isolamento & purificação
Expressão Gênica
Ralstonia/metabolismo
[Mh] Termos MeSH secundário: Proteínas de Bactérias/ultraestrutura
Canais de Cloreto/ultraestrutura
Cromatografia de Afinidade
Cromatografia em Gel
Microscopia Crioeletrônica
Detergentes/química
Escherichia coli/metabolismo
Glucosídeos/química
Proteínas de Fluorescência Verde/metabolismo
Maltose/análogos & derivados
Maltose/química
Espectrometria de Massas
Coloração Negativa
Estabilidade Proteica
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Chloride Channels); 0 (Detergents); 0 (Glucosides); 0 (dodecyl maltopyranoside); 0 (enhanced green fluorescent protein); 147336-22-9 (Green Fluorescent Proteins); 29836-26-8 (octyl-beta-D-glucoside); 69-79-4 (Maltose)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170925
[Lr] Data última revisão:
170925
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170711
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0180163



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