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1.
Chemosphere ; 362: 142787, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38972261

RESUMO

In this research, the dimensional catalysts of pure g-C3N4 photocatalysts (1D, 2D, and 3D) were investigated for the reduction of the highly toxic/carcinogenic Cr(VI) under visible light irradiation. The catalysts underwent explanation through various surface analysis techniques. According to the BET data, the specific surface area of the 3D catalyst was 1.3 and 7 times higher than those of the 2D and 1D CN catalysts, respectively. The 3D catalyst demonstrated superior performance, achieving an efficiency greater than 99% within 60 min under visible light irradiation in the presence of EDTA due to the abundance of active sites. The study also delved into the influence of factors such as the amount of EDTA-hole scavenger, pH, catalyst dosage, and temperature on the photocatalytic reduction of Cr(VI). Moreover, the 3D catalyst showed excellent reusability, maintaining an efficiency of more than 80% even after 10 cycles, and performed effectively in real water samples. The 3D CN catalyst, with its facile synthesis process, excellent visible light harvesting properties, high reduction efficiency that sustains over multiple cycles, and outstanding performance in real water samples, presents a significant advancement for practical applications in environmental remediation. This research contributes to a new understanding of developing efficient degradation methods for heavy metals in polluted water, highlighting the potential of 3D g-C3N4 catalysts in environmental cleanup efforts.

2.
Bioresour Technol ; 361: 127732, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35934247

RESUMO

In this study, the brown algae Sargassum Hemiphyllum was used as a carbon source for synthesis of magnetic porous biochar via pyrolyzing at high temperature and and doping iron oxide particles (Fe-BAB). Cu (II) species were removed from aqueous solutions using Fe-BAB under various conditions. Fe-BAB demonstrated superior Cu (II) adsorption (105.3 mg g-1) compared to other biochars. On the surface of Fe-BAB, there are several oxygen-containing functional groups, such as -COOH and -OH, which are likely responsible for the excellent heavy metal removal performance. By utilizing magnet, the Fe-BAB can be conveniently separated from the solution and ready for further usage. Multi-adsorption mechanisms were responsible for Cu adsorption on Fe-BAB. Using the magnetic algal biochar for heavy metal removal is feasible due to its high adsorption efficiency and simplicity of separation.


Assuntos
Metais Pesados , Sargassum , Poluentes Químicos da Água , Adsorção , Carvão Vegetal , Fenômenos Magnéticos , Poluentes Químicos da Água/análise
3.
Diagnostics (Basel) ; 12(2)2022 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-35204399

RESUMO

Chronic antibody-mediated rejection (AMR) is a key limiting factor for the clinical outcome of a kidney transplantation (Ktx), where early diagnosis and therapeutic intervention is needed. This study describes the identification of the biomarker CXC-motif chemokine ligand (CXCL) 9 as an indicator for AMR and presents a new aptamer-antibody-hybrid lateral flow assay (hybrid-LFA) for detection in urine. Biomarker evaluation included two independent cohorts of kidney transplant recipients (KTRs) from a protocol biopsy program and used subgroup comparisons according to BANFF-classifications. Plasma, urine and biopsy lysate samples were analyzed with a Luminex-based multiplex assay. The CXCL9-specific hybrid-LFA was developed based upon a specific rat antibody immobilized on a nitrocellulose-membrane and the coupling of a CXCL9-binding aptamer to gold nanoparticles. LFA performance was assessed according to receiver operating characteristic (ROC) analysis. Among 15 high-scored biomarkers according to a neural network analysis, significantly higher levels of CXCL9 were found in plasma and urine and biopsy lysates of KTRs with biopsy-proven AMR. The newly developed hybrid-LFA reached a sensitivity and specificity of 71% and an AUC of 0.79 for CXCL9. This point-of-care-test (POCT) improves early diagnosis-making in AMR after Ktx, especially in KTRs with undetermined status of donor-specific HLA-antibodies.

4.
ACS Chem Biol ; 17(1): 129-137, 2022 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-35018777

RESUMO

Renal rejection is a major incidence in patients after kidney transplantation and associated with allograft scarring and function loss, especially in antibody-mediated rejection. Regular clinical monitoring of kidney-transplanted patients is thus necessary, but measuring donor-specific antibodies is not always predictive, and graft biopsies are time-consuming and costly and may come up with a histological result unsuspicious for rejection. Therefore, a noninvasive diagnostic approach to estimate an increased probability of kidney graft rejection by measuring specific biomarkers is highly desired. The chemokine CXCL9 is described as an early indicator of rejection. In this work, we identified clickmers and an aptamer by split-combine click-SELEX (systematic evolution of ligands by exponential enrichment) that bind CXLC9 with high affinity. The aptamers recognize native CXCL9 and maintain binding properties under urine conditions. These features render the molecules as potential binding and detector probes for developing point-of-care devices, e.g., lateral flow assays, enabling the noninvasive monitoring of CXCL9 in renal allograft patients.


Assuntos
Quimiocina CXCL9/química , Química Click , Rejeição de Enxerto/metabolismo , Biomarcadores/metabolismo , Humanos , Ligantes , Ligação Proteica
5.
High Blood Press Cardiovasc Prev ; 28(2): 141-150, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33453048

RESUMO

INTRODUCTION: Modelling of associations of systolic blood pressure (BP) and blood glucose (BG) with their explanatory factors in separate regressions treats them as having independent biological mechanisms. This can lead to statistical inferences that are unreliable because the substantial overlap in their etiologic and disease mechanisms is ignored. AIM: This study aimed to examine the relationship of systolic blood pressure (BP) and blood glucose (BG) with measures of obesity and central fat distribution and other factors whilst taking account of the inter-dependence between them. METHODS: Participants (n = 14706, 53.5 % females) aged 25-64 years were selected by multi-stage stratified cluster sampling from eight provinces each representing one of the eight geographical regions of Vietnam. Measurements were made using the World Health Organization STEPS protocols. RESULTS: Structural modelling identified direct effects for BG (men P = 0.000, women P = 0.029), age (men P = 0.000, women P = 0.000) and body mass index (BMI) (men P = 0.000, women P = 0.000) in the estimation of systolic BP, and for systolic BP (men P = 0.036, women P = 0.000) and waist circumference (WC) (men P = 0.032, women P = 0.009) in the estimation of BG. There were indirect effects of age, cholesterol, physical activity and tobacco smoking via their influence on WC and BMI. The errors in estimation of systolic BP and BG were correlated (men P = 0.000, women P = 0.004), the stability indices (men 0.466, women 0.495) showed the non-recursive models were stable, and the proportion of variance explained was mid-range (men 0.553, women 0.579). CONCLUSION: This study provided statistical evidence of a feedback loop between systolic BP and BG. BMI and WC were confirmed to be their primary explanatory factors. Saturated fat intake and physical activity were identified as possible targets of intervention for overweight and obesity, and indirectly for reducing systolic BP and BG. Harmful/hazardous alcohol intake was identified as a target of intervention for systolic BP.


Assuntos
Adiposidade , Glicemia/metabolismo , Pressão Sanguínea , Transtornos do Metabolismo de Glucose/sangue , Hipertensão/fisiopatologia , Obesidade/fisiopatologia , Adulto , Biomarcadores/sangue , Índice de Massa Corporal , Feminino , Transtornos do Metabolismo de Glucose/diagnóstico , Transtornos do Metabolismo de Glucose/epidemiologia , Humanos , Hipertensão/diagnóstico , Hipertensão/epidemiologia , Masculino , Pessoa de Meia-Idade , Obesidade/diagnóstico , Obesidade/epidemiologia , Prognóstico , Medição de Risco , Fatores de Risco , Vietnã/epidemiologia , Circunferência da Cintura
6.
ACS Comb Sci ; 22(11): 617-629, 2020 11 09.
Artigo em Inglês | MEDLINE | ID: mdl-32894679

RESUMO

For improved cost-effectiveness and temperature-stability, a ready to use lateral flow assay (LFA) is developed in this work for detecting inflammation/infection biomarker C-reactive protein (CRP) in human patient samples on the basis of aptamers. In prescreening investigations, an aptamer with CRP affinity was immobilized on microarray chips in forward and sandwich formats to optimize assay conditions. We suggest these microarray techniques as a resource-sparing and fast-screening instrument for evaluation of various conditions. The capability of the aptamer to detect CRP was shown. Optimized assay conditions were consequently transferred to the LFA-platform. Here we could demonstrate for the first time an aptamer-based LFA for the detection of CRP in human patient samples in pathologically relevant concentrations. The cutoff for CRP detection is set at 10 mg/L, providing a distinctive "yes" (≥10 mg/L CRP) or "no" (<10 mg/L CRP) answer for the patient. The resulting aptamer-based LFA is promising with regard to its application as point-of-care testing (POCT) for efficient monitoring, especially of patients affected by frequent infections or inflammations.


Assuntos
Aptâmeros de Peptídeos/química , Proteína C-Reativa/análise , Citometria de Fluxo/métodos , Análise Serial de Proteínas/métodos , Sequência de Aminoácidos , Técnicas Biossensoriais , Proteína C-Reativa/metabolismo , Colódio/química , Ouro/química , Humanos , Nanopartículas Metálicas/química , Sensibilidade e Especificidade , Propriedades de Superfície
7.
Elife ; 92020 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-32840482

RESUMO

Avian influenza outbreaks have been occurring on smallholder poultry farms in Asia for two decades. Farmer responses to these outbreaks can slow down or accelerate virus transmission. We used a longitudinal survey of 53 small-scale chicken farms in southern Vietnam to investigate the impact of outbreaks with disease-induced mortality on harvest rate, vaccination, and disinfection behaviors. We found that in small broiler flocks (≤16 birds/flock) the estimated probability of harvest was 56% higher when an outbreak occurred, and 214% higher if an outbreak with sudden deaths occurred in the same month. Vaccination and disinfection were strongly and positively correlated with the number of birds. Small-scale farmers - the overwhelming majority of poultry producers in low-income countries - tend to rely on rapid sale of birds to mitigate losses from diseases. As depopulated birds are sent to markets or trading networks, this reactive behavior has the potential to enhance onward transmission.


The past few decades have seen the circulation of avian influenza viruses increase in domesticated poultry, regularly creating outbreaks associated with heavy economic loss. In addition, these viruses can sometimes 'jump' into humans, potentially allowing new diseases ­ including pandemics ­ to emerge. The Mekong river delta, in southern Vietnam, is one of the regions with the highest circulation of avian influenza. There, a large number of farmers practice poultry farming on a small scale, with limited investments in disease prevention such as vaccination or disinfection. Yet, it was unclear how the emergence of an outbreak could change the behavior of farmers. To learn more, Delabouglise et al. monitored 53 poultry farms, with fewer than 1000 chickens per farm, monthly for over a year and a half. In particular, they tracked when outbreaks occurred on each farm, and how farmers reacted. Overall, poultry farms with more than 17 chickens were more likely to vaccinate their animals and use disinfection practices than smaller farms. However, disease outbreaks did not affect vaccination or disinfection practices. When an outbreak occurred, farmers with fewer than 17 chickens tended to sell their animals earlier. For instance, they were 214% more likely to send their animals to market if an outbreak with sudden deaths occurred that month. Even if they do not make as much money selling immature individuals, this strategy may allow them to mitigate economical loss: they can sell animals that may die soon, saving on feeding costs and potentially avoiding further contamination. However, as animals were often sold alive in markets or to itinerant sellers, this practice increases the risk of spreading diseases further along the trade circuits. These data could be most useful to regional animal health authorities, which have detailed knowledge of local farming systems and personal connections in the communities where they work. This can allow them to effect change. They could work with small poultry farmers to encourage them to adopt efficient disease management strategies. Ultimately, this could help control the spread of avian influenza viruses, and potentially help to avoid future pandemics.


Assuntos
Criação de Animais Domésticos/estatística & dados numéricos , Surtos de Doenças , Fazendas/estatística & dados numéricos , Aves Domésticas , Animais , Surtos de Doenças/estatística & dados numéricos , Surtos de Doenças/veterinária , Desinfecção/estatística & dados numéricos , Fazendeiros , Humanos , Influenza Aviária , Estudos Longitudinais , Modelos Estatísticos , Rios , Vacinação/estatística & dados numéricos , Vacinação/veterinária , Vietnã
8.
Acta Crystallogr E Crystallogr Commun ; 75(Pt 12): 1861-1865, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31871746

RESUMO

In the title compound, C17H21NO4S, the 1,4-di-hydro-pyridine ring has an envelope conformation with the Csp 3 atom at the flap. The thio-phene ring is nearly perpendicular to the best plane through the 1,4-di-hydro-pyridine ring, the dihedral angle being 82.19 (13)°. In the crystal, chains running along the b-axis direction are formed through N-H⋯O inter-actions between the 1,4-di-hydro-pyridine N atom and one of the O atoms of the ester groups. Neighbouring chains are linked by C-H⋯O and C-H⋯π inter-actions. A Hirshfeld surface analysis shows that the most prominent contributuion to the surface contacts are H⋯H contacts (55.1%).

9.
Langmuir ; 35(42): 13761-13768, 2019 10 22.
Artigo em Inglês | MEDLINE | ID: mdl-31553189

RESUMO

Ultrafine bubbles exist in all liquids and are naturally stable. As their properties are not entirely known, it is unclear how they impact the surrounding solution and comparable-sized particles within it. It is essential to further investigate the properties of ultrafine bubbles in order to expand their industrial application. In this regard, the effect of ultrafine bubbles on bacterial development is of particular interest. Our current study, using optical density measurements and fluorescence microscopic images has demonstrated that ultrafine gas bubbles impact the morphology and phenotype of Escherichia coli and Pseudomonas aeruginosa. Specifically, Fourier transform infrared spectroscopic measurements indicated a thickening of bacterial membranes in samples exposed to ultrafine bubbles. The study also confirmed that ultrafine bubbles can inhibit bacterial cell growth. This study signifies the role of surface phenomena in bacterial culture, which is crucial in the upstream processes of recombinant DNA technology applications.


Assuntos
Membrana Celular/metabolismo , Escherichia coli/metabolismo , Pseudomonas aeruginosa/metabolismo , Membrana Celular/química , Escherichia coli/química , Microscopia de Fluorescência , Tamanho da Partícula , Pseudomonas aeruginosa/química
10.
Acta Crystallogr E Crystallogr Commun ; 75(Pt 7): 957-963, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31392003

RESUMO

The synthesis, spectroscopic data and crystal and mol-ecular structures of four 3-(3-phenyl-prop-1-ene-3-one-1-yl)thio-phene derivatives, namely 1-(4-hydroxy-phen-yl)-3-(thio-phen-3-yl)prop-1-en-3-one, C13H10O2S, (1), 1-(4-meth-oxy-phen-yl)-3-(thio-phen-3-yl)prop-1-en-3-one, C14H12O2S, (2), 1-(4-eth-oxy-phen-yl)-3-(thio-phen-3-yl)prop-1-en-3-one, C15H14O2S, (3), and 1-(4--bromophen-yl)-3-(thio-phen-3-yl)prop-1-en-3-one, C13H9BrOS, (4), are described. The four chalcones have been synthesized by reaction of thio-phene-3-carbaldehyde with an aceto-phenone derivative in an absolute ethanol solution containing potassium hydroxide, and differ in the substituent at the para position of the phenyl ring: -OH for 1, -OCH3 for 2, -OCH2CH3 for 3 and -Br for 4. The thio-phene ring in 4 was found to be disordered over two orientations with occupancies 0.702 (4) and 0.298 (4). The configuration about the C=C bond is E. The thio-phene and phenyl rings are inclined by 4.73 (12) for 1, 12.36 (11) for 2, 17.44 (11) for 3 and 46.1 (6) and 48.6 (6)° for 4, indicating that the -OH derivative is almost planar and the -Br derivative deviates the most from planarity. However, the substituent has no real influence on the bond distances in the α,ß-unsaturated carbonyl moiety. The mol-ecular packing of 1 features chain formation in the a-axis direction by O-H⋯O contacts. In the case of 2 and 3, the packing is characterized by dimer formation through C-H⋯O inter-actions. In addition, C-H⋯π(thio-phene) inter-actions in 2 and C-H⋯S(thio-phene) inter-actions in 3 contribute to the three-dimensional architecture. The presence of C-H⋯π(thio-phene) contacts in the crystal of 4 results in chain formation in the c-axis direction. The Hirshfeld surface analysis shows that for all four derivatives, the highest contribution to surface contacts arises from contacts in which H atoms are involved.

11.
BMC Vet Res ; 15(1): 205, 2019 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-31208467

RESUMO

BACKGROUND: Poultry farming is widely practiced by rural households in Vietnam and the vast majority of domestic birds are kept on small household farms. However, smallholder poultry production is constrained by several issues such as infectious diseases, including avian influenza viruses whose circulation remains a threat to public health. This observational study describes the demographic structure and dynamics of small-scale poultry farms of the Mekong river delta region. METHOD: Fifty three farms were monitored over a 20-month period, with farm sizes, species, age, arrival/departure of poultry, and farm management practices recorded monthly. RESULTS: Median flock population sizes were 16 for chickens (IQR: 10-40), 32 for ducks (IQR: 18-101) and 11 for Muscovy ducks (IQR: 7-18); farm size distributions for the three species were heavily right-skewed. Muscovy ducks were kept for long periods and outdoors, while chickens and ducks were farmed indoors or in pens. Ducks had a markedly higher removal rate (broilers: 0.14/week; layer/breeders: 0.05/week) than chickens and Muscovy ducks (broilers: 0.07/week; layer/breeders: 0.01-0.02/week) and a higher degree of specialization resulting in a substantially shorter life span. The rate of mortality due to disease did not differ much among species, with birds being less likely to die from disease at older ages, but frequency of disease symptoms differed by species. Time series of disease-associated mortality were correlated with population size for Muscovy ducks (Kendall's coefficient τ = 0.49, p-value < 0.01) and with frequency of outdoor grazing for ducks (τ = 0.33, p-value = 0.05). CONCLUSION: The study highlights some challenges to disease control in small-scale multispecies poultry farms. The rate of interspecific contact and overlap between flocks of different ages is high, making small-scale farms a suitable environment for pathogens circulation. Muscovy ducks are farmed outdoors with little investment in biosecurity and few inter-farm movements. Ducks and chickens are more at-risk of introduction of pathogens through movements of birds from one farm to another. Ducks are farmed in large flocks with high turnover and, as a result, are more vulnerable to disease spread and require a higher vaccination coverage to maintain herd immunity.


Assuntos
Criação de Animais Domésticos/métodos , Galinhas , Patos , Doenças das Aves Domésticas/epidemiologia , Fatores Etários , Animais , Fazendas/estatística & dados numéricos , Dinâmica Populacional , Doenças das Aves Domésticas/mortalidade , Doenças das Aves Domésticas/prevenção & controle , Doenças das Aves Domésticas/virologia , Vietnã
12.
PLoS Curr ; 92017 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-28736677

RESUMO

BACKGROUND: Subtype H5N1 avian influenza viruses, both high pathogenicity and low pathogenicity, have been enzootic in Vietnam since 2001.  The viruses are readily identified at live bird markets, but virus prevalence on smallholder poultry is typically zero or very low.  If the true direction of the viral transmission chain is farm to market, it is unknown why farm prevalence should be low when market prevalence is moderate to high. METHODS: We established a cohort of 50 smallholder poultry farms in Ca Mau province in the Mekong Delta region of Vietnam.  From March 2016 to January 2017, we collected naso-pharyngeal and cloacal samples from 156 ducks and 96 chickens.  In addition, 126 environmental samples were collected.  Samples were assayed for H5 subtype influenza by real-time RT-PCR. Results/Discussion: None of the 378 collected samples were positive for H5 influenza.  This is likely to mean that circulation of subtype H5 influenza viruses was low in Ca Mau in 2016.  Detection of avian influenza on smallholder poultry farms is necessary to determine the directionality and association between farm prevalence and market prevalence of avian influenza viruses.  Larger farm-level studies should be planned as these will be critical for determining the presence and strength of this association.

13.
J Heterocycl Chem ; 54(1): 155-160, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-28439141

RESUMO

Many 1,2,4-benzotriazine 1,4-dioxides display the ability to selectively kill the oxygen-poor cells found in solid tumors. As a result, there is a desire for synthetic routes that afford access to substituted 1,2,4-benzotriazine 1-oxides that can be used as direct precursors in the synthesis of 1,2,4-benzotriazine 1,4-dioxides. Here we describe the use of Suzuki-Miyaura and Buchwald-Hartwig cross-coupling reactions for the construction of various 1,2,4-benzotriazine 1-oxide analogs bearing substituents at the 3-, 6-, and 7-positions.

14.
Eng Life Sci ; 17(8): 953-958, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32624844

RESUMO

The selection of aptamers represents a promising route in the development of high affinity ligands. In these processes the formation of by-products is a common problem during the PCR-based amplification of complex oligonucleotide libraries. One approach to overcome this drawback is to separate each template oligonucleotide into an individual reaction compartment provided by a droplet. This method, termed emulsion PCR (ePCR), has already emerged to a standard method in sample preparation for 2nd generation sequencing. In this work, we compare different literature protocols that have been developed to generate stable emulsions for ePCR. We investigate different emulsification methods and evaluate the importance of the initial template concentration. We demonstrate that emulsion stability is of utmost importance for the successful inhibition of by-product formation and give an optimized protocol for generation of an emulsified PCR.

15.
Appl Ergon ; 58: 151-155, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27633208

RESUMO

Gavage is a common technique for orally administering compounds to small laboratory animals using a syringe. It involves highly repetitive thumb extensor exertions for filling the syringe, a risk factor for DeQuervain's tenosynovitis. As an intervention, a series of bench tests were performed varying fluid viscosity, syringe size and needle size to determine the forces required for drawing fluid. Forces up to 28 N were observed for a viscosity of 0.29 Pa s. A guide is presented to minimize thumb forces for a particular combination of syringe (3 mL, 5 mL and 10 mL), fluid viscosity (0.001 Pa s, 0.065 Pa s, 0.21 and 0.29 Pa s), and needle length (52 mm, 78 mm and 100 mm) based on maximum acceptable exertion levels. In general, a small syringe and large needle size had a greater number of acceptable rat gavages per day due to the lower forces experienced as compared to all other syringe and needle combinations.


Assuntos
Pesquisa Biomédica/instrumentação , Transtornos Traumáticos Cumulativos/prevenção & controle , Músculo Esquelético/lesões , Traumatismos Ocupacionais/prevenção & controle , Tenossinovite/prevenção & controle , Polegar , Administração Oral , Animais , Feminino , Humanos , Masculino , Agulhas , Azeite de Oliva , Pressão , Ratos , Seringas , Viscosidade , Água
16.
Front Microbiol ; 6: 677, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26236285

RESUMO

Metal and metal-oxide nanoparticles (NPs) are used in numerous applications and have high likelihood of entering engineered and natural environmental systems. Careful assessment of the interaction of these NPs with bacteria, particularly biofilm bacteria, is necessary. This perspective discusses mechanisms of NP interaction with bacteria and identifies challenges in understanding NP-biofilm interaction, considering fundamental material attributes and inherent complexities of biofilm structure. The current literature is reviewed, both for planktonic bacteria and biofilms; future challenges and complexities are identified, both in light of the literature and a dataset on the toxicity of silver NPs toward planktonic and biofilm bacteria. This perspective aims to highlight the complexities in such studies and emphasizes the need for systematic evaluation of NP-biofilm interaction.

17.
Biochemistry ; 48(32): 7673-85, 2009 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-19610625

RESUMO

The flavoprotein methylenetetrahydrofolate reductase from Escherichia coli catalyzes the reduction of 5,10-methylenetetrahydrofolate (CH(2)-H(4)folate) by NADH via a ping-pong reaction mechanism. Structures of the reduced enzyme in complex with NADH and of the oxidized Glu28Gln enzyme in complex with CH(3)-H(4)folate [Pejchal, R., Sargeant, R., and Ludwig, M. L. (2005) Biochemistry 44, 11447-11457] have revealed Phe223 as a conformationally mobile active site residue. In the NADH complex, the NADH adopts an unusual hairpin conformation and is wedged between the isoalloxazine ring of the FAD and the side chain of Phe223. In the folate complex, Phe223 swings out from its position in the NADH complex to stack against the p-aminobenzoate ring of the folate. Although Phe223 contacts each substrate in E. coli MTHFR, this residue is not invariant; for example, a leucine occurs at this site in the human enzyme. To examine the role of Phe223 in substrate binding and catalysis, we have constructed mutants Phe223Ala and Phe223Leu. As predicted, our results indicate that Phe223 participates in the binding of both substrates. The Phe223Ala mutation impairs NADH and CH(2)-H(4)folate binding each 40-fold yet slows catalysis of both half-reactions less than 2-fold. Affinity for CH(2)-H(4)folate is unaffected by the Phe223Leu mutation, and the variant catalyzes the oxidative half-reaction 3-fold faster than the wild-type enzyme. Structures of ligand-free Phe223Leu and Phe223Leu/Glu28Gln MTHFR in complex with CH(3)-H(4)folate have been determined at 1.65 and 1.70 A resolution, respectively. The structures show that the folate is bound in a catalytically competent conformation, and Leu223 undergoes a conformational change similar to that observed for Phe223 in the Glu28Gln-CH(3)-H(4)folate structure. Taken together, our results suggest that Leu may be a suitable replacement for Phe223 in the oxidative half-reaction of E. coli MTHFR.


Assuntos
Proteínas de Escherichia coli , Escherichia coli/enzimologia , Metilenotetra-Hidrofolato Redutase (NADPH2) , Fenilalanina/química , Fenilalanina/metabolismo , Conformação Proteica , Substituição de Aminoácidos , Animais , Domínio Catalítico , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Ácido Fólico/metabolismo , Humanos , Metilenotetra-Hidrofolato Redutase (NADPH2)/química , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Molecular , Mutagênese Sítio-Dirigida , NAD/metabolismo , Oxirredução , Fenilalanina/genética , Ligação Proteica
18.
J Mater Sci Mater Med ; 19(9): 2993-3003, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18360794

RESUMO

Hydroxyapatite (HA) nanoparticles have been studied as nano-sized carriers for the delivery of therapeutic agents. One important consideration for these carriers to be used effectively is their bio-distribution in vivo, of which particle size has a significant effect. In this work, HA nanoparticles doped with Ethylene-diamine-tetramethylene-phosphonate (EDTMP) were synthesized via co-precipitation as a model for HA doped with (153)Samarium ((153)Sm) EDTMP. EDTMP has high affinity for radioactive (153)Sm isotopes that can emit both gamma and beta radiation. The effects of synthesis temperature, amount of dopant and hydrothermal treatment on the size of HA-EDTMP nanoparticles were therefore studied. The results showed that the EDTMP ligand was successfully incorporated in the nanoparticles without changing the crystal structure as shown from X-ray diffractometer (XRD) analysis. From the Field Emission Scanning Electron Microscopy (FESEM) and Transmission Electron Microscopy (TEM) micrographs, it was observed that shorter rod-like nanoparticles, obtained at low synthesis temperatures, became elongated needle-like nanoparticles with increasing temperature. Increasing dopant amount by five fold increases particle size slightly, while a two fold increase in dopant amount has no significant effect. Hydrothermal treatment increases particle crystallinity and results in smooth elongated rod-like structures. The size of HA nanoparticles doped with EDTMP can therefore be manipulated by controlling synthesis temperature and through hydrothermal treatment.


Assuntos
Durapatita/química , Compostos Organometálicos/química , Compostos Organofosforados/química , Radioisótopos/química , Samário/química , Analgésicos não Narcóticos/química , Partículas beta , Raios gama , Ligantes , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Modelos Teóricos , Nanopartículas/química , Nanotecnologia/métodos , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
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