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1.
Yakugaku Zasshi ; 140(10): 1243-1249, 2020.
Artigo em Japonês | MEDLINE | ID: mdl-32999203

RESUMO

Here the author describes the tumor-selective delivery of a fluorescence photosensitizing agent and an antitumor agent, based on the polymer effect of an N-(2-hydroxypropyl)methacrylamide (HPMA) based copolymer, by utilizing the enhanced permeability and retention (EPR) effect seen in solid tumors. Firstly, the tumor distribution of the photosensitizer, zinc-protoporphyrin IX (ZnPP), was significantly increased by conjugation with the HPMA polymer (P-ZnPP). The P-ZnPP suppressed tumor growth by local generation of cytotoxic singlet oxygen, and the tumor tissue was visualized by fluorescence upon light irradiation. Subsequently, a two-step mechanism for tumor selectivity was observed for the cytotoxic anthracycline, pirarubicin (THP), which conjugated the HPMA-based copolymer via a hydrazone bond (P-THP). The EPR-dependent accumulation of P-THP and the tumor-selective release of THP in the tumor tissues led to highly tumor-selective toxicity. Rapid cell uptake of THP compared to other anthracyclines, and deeper P-THP penetration of the tumor cell spheroid were attributed to the superior antitumor activity of P-THP. The molecular weight of P-THP affected its antitumor activity; oligomeric P-THP derivatives with higher molecular weights, DP-THP and SP-THP, showed even higher antitumor activity. P-THP was effective for both implanted tumor and autochthonous tumor models. These results indicate that nano-sized anticancer drugs based on polymer effect are promising clinical therapeutics.


Assuntos
Antineoplásicos , Sistemas de Liberação de Medicamentos , Desenvolvimento de Medicamentos/métodos , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Fármacos Fotossensibilizantes , Polímeros , Protoporfirinas , Animais , Antraciclinas/química , Antraciclinas/metabolismo , Antineoplásicos/química , Antineoplásicos/metabolismo , Doxorrubicina/análogos & derivados , Doxorrubicina/química , Doxorrubicina/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Metacrilatos/química , Terapia de Alvo Molecular , Peso Molecular , Neoplasias/patologia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/metabolismo , Protoporfirinas/química , Protoporfirinas/metabolismo
2.
J Environ Qual ; 49(1): 172-183, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33016355

RESUMO

Use of recycled concrete aggregate (RCA) as highway basecourse material conserves virgin aggregate, reduces energy consumption and CO2 emissions, and may also decrease costs during construction. However, concerns remain over possible negative environmental impacts associated with high pH (>11) effluent from RCA in contact with water. This study examines the reactive transport of high-pH and high-alkalinity water, modeled on RCA leachate, through model subgrade soils. By developing an understanding of the reactions controlling effluent neutralization, this study aims to quantify the change in pH from the discharge site through surrounding subgrade soils. Four types of subgrade soils with a range of mineral composition, Atterberg limits, and cation exchange capacities (CECs) are examined. They include a clayey sand (SC10), low-plasticity clays (M14, SC25), and a high-plasticity clay (CH38). Batch reaction experiments are used to develop kinetic parameters describing the neutralization of high-pH and -alkalinity leachate by clay minerals through mineral dissolution and reprecipitation. Given this information, a reactive transport model incorporating advection, diffusion, and reaction is used to model the change in pH as a function of distance traveled through model subgrade soils and is applied to laboratory-scale column experiments. The rate at which the high pH front travels is directly related to a soil's clay mineral content. Soils with high CECs effectively delay the propagation of hydroxide front by the dissolution of clay minerals. This study demonstrates that common subgrade soils with moderate clay content will effectively neutralize high pH leachate initially produced by RCA.


Assuntos
Poluentes do Solo/análise , Solo , Argila , Concentração de Íons de Hidrogênio , Reciclagem
3.
J Environ Qual ; 49(5): 1445-1452, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33016443

RESUMO

Volatilization of NH3 following urea application or livestock urine deposition can result in significant loss of N to the environment. Urea hydrolysis to NH4 + results in an increase in pH, which in turn promotes transformation of NH4 + to NH3 . Accurately predicting changes in soil pH following urea (or urine) application will allow successful simulation of NH3 volatilization. The magnitude of the pH change depends on the soil's pH buffering capacity (pHBC). However, as actual pHBC values are not generally available, pHBC proxies (e.g., cation exchange capacity) have been used in modeling studies. In a 34-d laboratory incubation study, we measured soil pH and mineral N (NH4 + and NO3 - ) following a large application of urea (800 mg N kg-1 soil) to four soils with a range of pHBC values. In a second incubation, pH changes and mineral N dynamics were monitored in soil treated with sheep urine (773 mg N kg-1 soil) in the absence and presence of the nitrification inhibitor dicyandiamide. In both incubations, pH changes associated with urea hydrolysis and subsequent nitrification of NH4 + were predicted well using measured pHBC data. Our results confirmed that pHBC is base-type dependent (values greater when measured using KOH than NH4 OH). Soil pHBC is easily measured, and the use of a measured value (determined using NH4 OH) can improve model simulations of pH in the field and, potentially, lead to improved estimates of NH3 loss from animal-deposited urine patches and urea-treated soil.


Assuntos
Solo , Ureia , Animais , Concentração de Íons de Hidrogênio , Nitrificação , Ovinos , Volatilização
4.
J Environ Qual ; 49(4): 961-972, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33016495

RESUMO

The effects of enhanced acid deposition from the atmosphere, and associated elevated inputs of N, are widely evident, especially for forests where excess N has led to a variety of deleterious effects. These include declines in biodiversity, a response that will likely require considerable time for recovery. The purpose of this study was to determine responses of plant nutrient availability in surface mineral soil to 25 yr of experimental acidification and N addition in a central Appalachian hardwood forest ecosystem. We hypothesized that chronic additions of (NH4 )2 SO4 will increase mineral N, decrease soil pH, P, and base cations, increase micronutrients (Mn2+ and Fe2+ ), and increase levels of Al3+ . Results supported these predictions, although Mn2+ did not vary significantly. Earlier work on these plots found no response of any of the extractable nutrients to 3 yr of treatment, yet after 25 yr, our results suggest that impacts are apparent in the top 5 cm of the A horizon. We surmise that impacts in these soils may have lagged behind the onset of acidification treatments or that several years of treatment were required to overcome preexisting differences in soil ions. Generally, current findings confirm that (NH4 )2 SO4 treatments have lowered the pH, enhanced levels of exchangeable Al3+ , and increased stream-water exports of NO3 - and base cations-a process that further acidifies soil. The combination of these changes in surface soils, with their high proportion of fine roots, may contribute to the reduced growth and competitiveness of some hardwood species at the acidified site.


Assuntos
Ecossistema , Solo , Região dos Apalaches , Florestas , Concentração de Íons de Hidrogênio
5.
Water Sci Technol ; 82(5): 887-904, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33031068

RESUMO

The effectiveness of two Azadirachta indica bark activated carbons (ABAC) for the removal of selected toxic metals from mining wastewater and the attendant challenge of multivariate factors in the process were enhanced through optimization studies. Experimental design was carried out using adsorbent dosage, agitation rate, contact time, grain size, pH and temperature as independent variables. Batch adsorption experiments were conducted using the experimental design result, then the experimental data obtained were optimized using Design-Expert software and the results validated. Optimum values for ABAC-NaOH adsorbent were 1.999 g of adsorbent dosage, 149.73 rpm agitation rate, 119.55 min contact time, 2 mm grain size, pH of 7 and 30 °C temperature; while for ABAC-HCl adsorbent the optimum values were 3.993 g of adsorbent dosage, 150 rpm agitation rate, 120 min contact time, 2 mm grain size, pH of 7.001 and 30 °C temperature. These resulted in 100% removal efficiencies for all the selected toxic metals with standard errors of between 0.02 and 2.72%. So the optimization process is a very useful tool in adsorption studies. It has the merits of being economical, energy and time saving, and is therefore strongly recommended for the biosorption of toxic metals from mining wastewater using Azadirachta indica adsorbent.


Assuntos
Azadirachta , Águas Residuárias , Concentração de Íons de Hidrogênio , Metais , Mineração , Casca de Planta
6.
J Environ Manage ; 263: 110415, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32883481

RESUMO

Drinking water containing excess fluoride is a major health concern across the globe. The present study reports the feasibility of zirconium impregnated hybrid anion exchange resin (HAIX-Zr) for treating fluoride contaminated groundwater. The HAIX-Zr resin was prepared by impregnating ZrO2 nanoparticles on polymeric anion exchanger resin. Fluoride uptake by HAIX-Zr was quite rapid, 60% removal was obtained within 30 min. Kinetics of fluoride uptake by HAIX-Zr resin followed the pseudo-second-order kinetic model and adsorption data fitted best to Freundlich adsorption isotherm model. Maximum fluoride uptake capacity was observed as 12.0 mg/g. The defluoridation capacity of the resin decreases with increase in solution pH. The co-existing anions like chloride, phosphate, bicarbonate, nitrate, and sulphate at 100 mg/L concentration significantly affected fluoride removal and bicarbonate showed the highest interference. Continuous flow packed bed experiments were performed with real groundwater. To maintain a lower pH, weak acid cation exchange resin (INDION-236) was used before HAIX-Zr. It was observed that reducing the pH of the sample water to 4-4.5, increased the number of treated bed volumes fifteen times. Regeneration of fluoride-containing resin was done by passing 3% NaOH and 3% NaCl solution through an exhausted resin bed. The results revealed that HAIX-Zr can effectively remove fluoride from groundwater.


Assuntos
Água Subterrânea , Poluentes Químicos da Água/análise , Purificação da Água , Adsorção , Resinas de Troca de Ânions , Fluoretos , Concentração de Íons de Hidrogênio , Cinética , Zircônio
7.
Mar Environ Res ; 160: 104987, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32907725

RESUMO

Ocean acidification alters physiology, acid-base balance and metabolic activity in marine animals. Near future elevated pCO2 conditions could be expected to influence the bioaccumulation of metals, feeding rate and immune parameters in marine mussels. To better understand such impairments, a series of laboratory-controlled experiment was conducted by using a model marine mussel, Mytilus galloprovincialis. The mussels were exposed to three pH conditions according to the projected CO2 emissions in the near future (one ambient: 8.10 and two reduced: 7.80 and 7.50). At first, the bioconcentration of Ag and Cd was studied in both juvenile (2.5 cm) and adult (5.1 cm) mussels by using a highly sensitive radiotracer method (110mAg and 109Cd). The uptake and depuration kinetics were followed 21 and 30 days, respectively. The biokinetic experiments demonstrated that the effect of ocean acidification on bioconcentration was metal-specific and size-specific. The uptake, depuration and tissue distribution of 110mAg were not affected by elevated pCO2 in both juvenile and adult mussels, whereas 109Cd uptake significantly increased with decreasing pH in juveniles but not in adults. Regardless of pH, 110mAg accumulated more efficiently in juvenile mussels than adult mussels. After executing the biokinetic experiment, the perturbation was sustained by using the same mussels and the same experimental set-up, which enabled us to determine filtration rate, haemocyte viability, lysosomal membrane stability, circulating cell-free nucleic acids (ccf-NAs) and protein (ccf-protein) levels. The filtration rate and haemocyte viability gradually decreased by increasing pCO2 level, whereas the lysosomal membrane stability, ccf-NAs, and ccf-protein levels remained unchanged in the mussels exposed to elevated pCO2 for eighty-two days. This study suggests that acidified seawater partially shift metal bioaccumulation, physiological and cellular parameters in the mussel Mytilus galloprovincialis.


Assuntos
Dióxido de Carbono , Metais , Mytilus , Poluentes Químicos da Água , Animais , Bioacumulação , Concentração de Íons de Hidrogênio , Metais/farmacocinética , Mytilus/química , Água do Mar , Poluentes Químicos da Água/farmacocinética
8.
J Chem Phys ; 153(11): 115101, 2020 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-32962355

RESUMO

Broad-spectrum antiviral drugs are urgently needed to stop the Coronavirus Disease 2019 pandemic and prevent future ones. The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is related to the SARS-CoV and Middle East respiratory syndrome coronavirus (MERS-CoV), which have caused the previous outbreaks. The papain-like protease (PLpro) is an attractive drug target due to its essential roles in the viral life cycle. As a cysteine protease, PLpro is rich in cysteines and histidines, and their protonation/deprotonation modulates catalysis and conformational plasticity. Here, we report the pKa calculations and assessment of the proton-coupled conformational dynamics of SARS-CoV-2 in comparison to SARS-CoV and MERS-CoV PLpros using the recently developed graphical processing unit (GPU)-accelerated implicit-solvent continuous constant pH molecular dynamics method with a new asynchronous replica-exchange scheme, which allows computation on a single GPU card. The calculated pKa's support the catalytic roles of the Cys-His-Asp triad. We also found that several residues can switch protonation states at physiological pH among which is C270/271 located on the flexible blocking loop 2 (BL2) of SARS-CoV-2/CoV PLpro. Simulations revealed that the BL2 can open and close depending on the protonation state of C271/270, consistent with the most recent crystal structure evidence. Interestingly, despite the lack of an analogous cysteine, BL2 in MERS-CoV PLpro is also very flexible, challenging a current hypothesis. These findings are supported by the all-atom fixed-charge simulations and provide a starting point for more detailed studies to assist the structure-based design of broad-spectrum inhibitors against CoV PLpros.


Assuntos
Antivirais/farmacologia , Betacoronavirus/enzimologia , Desenho de Fármacos , Coronavírus da Síndrome Respiratória do Oriente Médio/enzimologia , Simulação de Dinâmica Molecular , Papaína/química , Papaína/metabolismo , Prótons , Sequência de Aminoácidos , Histidina , Concentração de Íons de Hidrogênio , Papaína/antagonistas & inibidores , Domínios Proteicos
9.
Chemosphere ; 258: 127213, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32947655

RESUMO

Worldwide, the aquatic environment is contaminated by micro-pollutants, such as ingredients of personal care products, pesticides and pharmaceuticals. This contamination is one of the major environmental issues of global concern. Adsorption is one of approach, which has been most extensively discussed within recent years for the reduction of the input of micro-pollutants into the environment. In the present study, the natural clay classified as Na-montmorillonite, was characterized and tested for its potential to remove four model compounds representing different polarity and ionizability: i) diatrizoic acid (DAT), ii) iopamidol (IOP), iii) metformin (MTF), and iv) carbamazepine (CBZ). The adsorption efficiency of clay was evaluated by initial compound concentration, effect of pH, contact time and temperature. The results indicated that clay was able to remove the pharmaceuticals from aqueous medium with an efficiency of 70% for CBZ and MTF. In contrast, clay showed a lower removal of 30% for DAT and no removal for IOP. The results indicate that clay could rapidly and efficiently reduce the concentration of CBZ and MTF, which could provide a solution to remove some substances, without undesirable by-product generation. However, this study clearly demonstrated that removal rates strongly depend on the compound. Albeit chemical structure may play a role for the different degree of removal, this study could not completely explain the sorption mechanism between sorbent-sorbate interactions.


Assuntos
Bentonita/química , Eliminação de Resíduos Líquidos/métodos , Poluentes Químicos da Água/análise , Adsorção , Carbamazepina , Argila , Concentração de Íons de Hidrogênio , Temperatura , Águas Residuárias
10.
Chemosphere ; 258: 127393, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32947669

RESUMO

UV/chlorine and chlorination processes have drawn great interests of water treatment utilities for oxidation and disinfection purposes. This work proposed a restricted chlorine-dosing strategy for UV/chlorine and post-chlorination under different pH and UV irradiation conditions by comprehensively assessing the oxidation of natural organic matter (NOM), formation of 9 haloacetic acids (HAA9) and bromate, and alteration of toxicity. During UV/chlorine with restricted chlorine doses, the oxidation of NOM chromophores (i.e., ΔUVA254) showed an apparent dependence on cumulative exposures of free available chlorine (CTFAC); Meanwhile, HAA9 formation was determined by CTFAC values and could be linearly correlated with ΔUVA254 irrespective of pH and UV irradiation wavelength. Irradiated by 254 nm LP-Hg lamp, the faster chlorine photolysis produced relatively higher steady-state concentrations of Cl• and HO• species but resulted in lower CTFAC. Reducing CTFAC values by operation parameters (pH, UV wavelength and irradiation fluence) could mitigate HAA9 formation during UV/chlorine at a specific chlorine dose. Additionally, high bromide concentration and acidic pH promoted more bromo-HAAs formation, and the presence of NOM significantly suppressed bromate formation. Analogous to ozonation, the UV/chlorine pre-oxidation could reduce the HAA9 formation potentials during post-chlorination at mildly alkaline pH. The photobacterium bioassay further demonstrated that although the UV/chlorine treatment might have increased the acute toxicity, the post-chlorination treatment could polish the acute toxicity to the level of chlorination alone. These results suggest that with the restricted chlorine-dosing strategy, the trade-off between oxidation/disinfection efficiency and DBPs formation can be controlled by monitoring CTFAC and ΔUVA254 values during UV/chlorine treatment.


Assuntos
Purificação da Água/métodos , Bromatos , Brometos/efeitos da radiação , Cloro , Desinfecção , Halogenação , Concentração de Íons de Hidrogênio , Oxirredução , Fotólise , Raios Ultravioleta , Poluentes Químicos da Água/análise , Purificação da Água/normas
11.
Pestic Biochem Physiol ; 170: 104704, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32980065

RESUMO

Carboxylesterases have widely been used in a series of industrial applications, especially, the detoxification of pesticide residues. In the present study, EstC, a novel carboxylesterase from Streptomyces lividans TK24, was successfully heterogeneously expressed, purified and characterized. Phylogenetic analysis showed that EstC can be assigned as the first member of a novel family XIX. Multiple sequence alignment indicated that EstC has highly conserved structural features, including a catalytic triad formed by Ser155, Asp248 and His278, as well as a canonical Gly-His-Ser-Ala-Gly pentapeptide. Biochemical characterization indicated that EstC exhibited maximal activity at pH 9.0 (Tris-HCl buffer) and 55 °C. It also showed higher activity towards short-chain substrates, with the highest activity for p-nitrophenyl acetate (pNPA2) (Km = 0.31 ± 0.02 mM, kcat/Km = 1923.35 ± 9.62 s-1 mM-1) compared to other pNP esters used in this experiment. Notably, EstC showed hyper-thermostability and good alkali stability. The activity of EstC had no significant changes when it was incubated under 55 °C for 100 h and reached half-life after incubation at 100 °C for 8 h. Beyond that, EstC also showed stability at pH ranging from 6.0 to 11.0 and about 90% residual activity still reserved after treatment at pH 8.0 or 9.0 for 26 h, especially. Furthermore, EstC had outstanding potential for bioremediation of chlorpyrifos-contaminated environment. The recombinant enzyme (0.5 U mL-1) could hydrolyze 79.89% chlorpyrifos (5 mg L-1) at 37 °C within 80 min. These properties will make EstC have a potential application value in various industrial productions and detoxification of chlorpyrifos residues.


Assuntos
Carboxilesterase/genética , Clorpirifos , Sequência de Aminoácidos , Hidrolases de Éster Carboxílico/genética , Clonagem Molecular , Concentração de Íons de Hidrogênio , Filogenia , Proteínas Recombinantes/genética , Especificidade por Substrato , Temperatura
12.
Nan Fang Yi Ke Da Xue Xue Bao ; 40(7): 972-980, 2020 Jul 30.
Artigo em Chinês | MEDLINE | ID: mdl-32895149

RESUMO

OBJECTIVE: To investigate the effects of acid-sensing ion channels (ASICs) on electrophysiological epileptic activities of mouse hippocampal pyramidal neurons in the extracellular acidotic condition. METHODS: We investigated effects of extracellular acidosis on epileptic activities induced by elevated extracellular K + concentration or the application of an antagonist of GABAA receptors in perfusate of mouse hippocampal slices under field potential recordings. We also tested the effects of extracellular acidosis on neuronal excitability under field potential recording and evaluated the changes in epileptic activities of the neurons in response to pharmacological inhibition of ASICs using a specific inhibitor of ASICs. RESULTS: Extracellular acidosis significantly suppressed epileptic activities of the hippocampal neurons by converting ictal-like epileptic activities to non-ictal-like epileptic activities in both high [K +]o and disinhibition models, and also suppressed the intrinsic excitability of the neurons. ASICs inhibitor did not antagonize the inhibitory effect of extracellular acidosis on ictal epileptic activities and intrinsic neuronal excitability, but exacerbated non-ictal epileptic activities of the neurons in extracellular acidotic condition in both high [K+]o and disinhibition models. CONCLUSIONS: ASICs can differentially modulate ictal-like and non-ictallike epileptic activities via its direct actions on excitatory neurons.


Assuntos
Acidose , Canais Iônicos Sensíveis a Ácido , Animais , Hipocampo , Concentração de Íons de Hidrogênio , Camundongos , Células Piramidais
13.
Nan Fang Yi Ke Da Xue Xue Bao ; 40(6): 843-849, 2020 Jun 30.
Artigo em Chinês | MEDLINE | ID: mdl-32895202

RESUMO

OBJECTIVE: To develop a fast, sensitive and cost-effective method based on resonance light scattering (RLS) for characterization of protein solubility to facilitate detection of changes in solubility of mutant proteins. METHODS: We examined the response curve of RLS intensities to the protein concentrations in synchronous scanning mode. The curve intersection points were searched to predict the maximal concentrations of the protein in dispersion state, which defined the solubility of the protein in this given state. Bovine serum albumin (BSA, 0-50 g/L) was used as the model to investigate the influences of pH values (6.5, 7.0, and 7.4) and salt concentrations (0.05, 0.10, 0.15, and 0.20 mol/L) on the determined solubility. The solubility of glutathione S-transferase isoenzymes alpha (GSTA, 0-27.0 g/L) and Mµ (GSTM, 0-20.0 g/L) were estimated for comparison. The RLS-based method was used to determine the solubility of uricase (MGU, 0-0.4 g/L) to provide assistance in improving the solubility of its mutants. RESULTS: We identified two intersection points in the RLS response curves of the tested proteins, among which the lower one represented an approximation of the maximal concentration (or the solubility of the protein) in single molecular dispersion, and the higher one the saturated concentration of the protein in multiple molecular aggregation. In HEPES buffer, the two intersection points of BSA (isoelectric point 4.6) both increased with the increase of pH (6.5-7.4), and their values were ~1.2 g/L and ~33 g/L at pH 7.4, respectively; the latter concentration approached the solubility of commercial BSA in the same buffer at the same pH. The addition of NaCl reduced the values of the two intersection points, and increasing salt ion concentration decreased the values of the lower intersection points. Further characterizations of GSTA and GSTM showed that the low concentration intersection points of the two proteins were ~0.7 g/L and ~0.8 g/L, and their high concentration intersection points were ~10 g/L and ~11 g/L, respectively, both lower than those of BSA, indicating the feasibility of the direct characterization of protein solubility by RLS. The two concentration intersection points of MGU were 0.24 g/L and 0.30 g/L, respectively, and the low concentration intersection point of its selected mutant was increased by 2 times. CONCLUSIONS: RLS allows direct characterization of the solubility of macromolecular proteins. This method, which is simple and sensitive and needs only a small amount of proteins, has a unique advantage for rapid comparison of solubility of low-abundance protein mutants.


Assuntos
Luz , Concentração de Íons de Hidrogênio , Espalhamento de Radiação , Solubilidade , Análise Espectral
14.
Acta Odontol Latinoam ; 33(2): 82-89, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32920609

RESUMO

The aim of this study was to evaluate the degree of conversion, cytotoxicity, solubility and pH of photopolymerizable calciumbased cements submitted to preheating. The degree of conversion was analyzed by Fourier transform infrared, cytotoxicity by the MTT test and solubility through loss of mass. The data were subjected to statistical tests (ANOVA / Tukey's, p<0.05). The photopolymerizable materials showed a low degree of conversion, regardless of preheating. All materials caused a reduction in cell viability at 24 hours and 7 days, with the Dycal (control) being more cytotoxic. Heat had a positive effect on Biocal at 7 days. Dycal is the most soluble material. Heat had no effect on the solubility or pH of the polymerizable materials. It is concluded that photopolymerizable calcium-based cements have a low degree of conversion and are soluble, which results in mild to moderate cytotoxicity.


Assuntos
Hidróxido de Cálcio/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Cimentos Dentários/química , Agentes de Capeamento da Polpa Dentária e Pulpectomia/toxicidade , Cálcio , Hidróxido de Cálcio/química , Cimentos Dentários/toxicidade , Capeamento da Polpa Dentária , Humanos , Concentração de Íons de Hidrogênio , Cura Luminosa de Adesivos Dentários , Processos Fotoquímicos , Polimerização , Agentes de Capeamento da Polpa Dentária e Pulpectomia/química
15.
Ecotoxicol Environ Saf ; 203: 111007, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32888586

RESUMO

Soil acidification is one of the crucial global environmental problems, affecting sustainable land use, crop yield, and ecosystem stability. Previous research reported the tolerance of crops to acid soil stress. However, the molecular response of woody plant to acid conditions remains largely unclear. Rhododendron L. is a widely distributed woody plant genus and prefers to grow in acidic soils. Herein, weighted gene coexpression network analysis was performed on R. protistum var. giganteum seedlings subjected to five pH treatments (3.5, 4.5, 5.5, 6.0, 7.0), and their ecophysiological characteristics were determined for the identification of their molecular responses to acidic environments. Through pairwise comparison, 855 differentially expressed genes (DEGs) associated with photosynthesis, cell wall, and phenylpropanoid metabolism were identified. Most of the DEGs related to photosynthesis and cell wall were up-regulated after pH 4.5 treatment. Results implied that the species improves its photosynthetic abilities and changes its cell wall characteristics to adapt to acidic conditions. Weighted gene co-expression network analyses showed that most of the hub genes were annotated to the biosynthetic pathways of ribosomal proteins and photosynthesis. Expression pattern analysis showed that genes encoding subunit ribosomal proteins decreased at pH 7.0 treatment, suggesting that pH 7.0 treatment led to cell injury in the seedlings. The species regulates protein synthesis in response to high pH stress (pH 7.0). The present study revealed the molecular response mechanism of woody plant R. protistum var. giganteum to acid environments. These findings can be useful in enriching current knowledge of how woody species adapt to soil acidification under global environmental changes.


Assuntos
Ácidos/farmacologia , Parede Celular/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Fotossíntese/efeitos dos fármacos , Madeira/efeitos dos fármacos , Parede Celular/genética , Parede Celular/metabolismo , Perfilação da Expressão Gênica , Redes Reguladoras de Genes/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Fotossíntese/genética , Plântula/efeitos dos fármacos , Plântula/metabolismo , Solo/química , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética , Madeira/genética , Madeira/metabolismo
16.
Proc Natl Acad Sci U S A ; 117(39): 24450-24458, 2020 09 29.
Artigo em Inglês | MEDLINE | ID: mdl-32900935

RESUMO

The current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has had an enormous impact on society worldwide, threatening the lives and livelihoods of many. The effects will continue to grow and worsen if economies begin to open without the proper precautions, including expanded diagnostic capabilities. To address this need for increased testing, we have developed a sensitive reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay compatible with current reagents, which utilizes a colorimetric readout in as little as 30 min. A rapid inactivation protocol capable of inactivating virions, as well as endogenous nucleases, was optimized to increase sensitivity and sample stability. This protocol, combined with the RT-LAMP assay, has a sensitivity of at least 50 viral RNA copies per microliter in a sample. To further increase the sensitivity, a purification protocol compatible with this inactivation method was developed. The inactivation and purification protocol, combined with the RT-LAMP assay, brings the sensitivity to at least 1 viral RNA copy per microliter in a sample. This simple inactivation and purification pipeline is inexpensive and compatible with other downstream RNA detection platforms and uses readily available reagents. It should increase the availability of SARS-CoV-2 testing as well as expand the settings in which this testing can be performed.


Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Pneumonia Viral/diagnóstico , Betacoronavirus/genética , Técnicas de Laboratório Clínico/economia , Colorimetria , Infecções por Coronavirus/economia , Infecções por Coronavirus/virologia , Genoma Viral/genética , Humanos , Concentração de Íons de Hidrogênio , Técnicas de Diagnóstico Molecular/economia , Técnicas de Amplificação de Ácido Nucleico/economia , Pandemias , Pneumonia Viral/virologia , Estabilidade de RNA , RNA Viral/química , RNA Viral/genética , RNA Viral/isolamento & purificação , Sensibilidade e Especificidade , Fatores de Tempo , Proteínas Virais/genética , Inativação de Vírus
17.
Nat Commun ; 11(1): 4864, 2020 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-32978392

RESUMO

The synthesis of customized glycoconjugates constitutes a major goal for biocatalysis. To this end, engineered glycosidases have received great attention and, among them, thioglycoligases have proved useful to connect carbohydrates to non-sugar acceptors. However, hitherto the scope of these biocatalysts was considered limited to strong nucleophilic acceptors. Based on the particularities of the GH3 glycosidase family active site, we hypothesized that converting a suitable member into a thioglycoligase could boost the acceptor range. Herein we show the engineering of an acidophilic fungal ß-xylosidase into a thioglycoligase with broad acceptor promiscuity. The mutant enzyme displays the ability to form O-, N-, S- and Se- glycosides together with sugar esters and phosphoesters with conversion yields from moderate to high. Analyses also indicate that the pKa of the target compound was the main factor to determine its suitability as glycosylation acceptor. These results expand on the glycoconjugate portfolio attainable through biocatalysis.


Assuntos
Tolerância a Medicamentos/fisiologia , Fungos/enzimologia , Fungos/metabolismo , Xilosidases/química , Xilosidases/metabolismo , Biocatálise , Domínio Catalítico , Fungos/efeitos dos fármacos , Glicoconjugados/metabolismo , Glicosídeo Hidrolases/metabolismo , Glicosídeos/química , Glicosilação , Concentração de Íons de Hidrogênio , Cinética , Modelos Moleculares , Mutagênese , Especificidade por Substrato , Talaromyces/enzimologia , Talaromyces/genética , Xilosidases/genética
18.
Proc Biol Sci ; 287(1934): 20201506, 2020 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-32900308

RESUMO

The sea urchin embryo develops a calcitic endoskeleton through intracellular formation of amorphous calcium carbonate (ACC). Intracellular precipitation of ACC, requires [Formula: see text] concentrating as well as proton export mechanisms to promote calcification. These processes are of fundamental importance in biological mineralization, but remain largely unexplored. Here, we demonstrate that the calcifying primary mesenchyme cells (PMCs) use Na+/H+-exchange (NHE) mechanisms to control cellular pH homeostasis during maintenance of the skeleton. During skeleton re-calcification, pHi of PMCs is increased accompanied by substantial elevation in intracellular [Formula: see text] mediated by the [Formula: see text] cotransporter Sp_Slc4a10. However, PMCs lower their pHi regulatory capacities associated with a reduction in NHE activity. Live-cell imaging using green fluorescent protein reporter constructs in combination with intravesicular pH measurements demonstrated alkaline and acidic populations of vesicles in PMCs and extensive trafficking of large V-type H+-ATPase (VHA)-rich acidic vesicles in blastocoelar filopodial cells. Pharmacological and gene expression analyses underline a central role of the VHA isoforms Sp_ATP6V0a1, Sp_ATP6V01_1 and Sp_ATPa1-4 for the process of skeleton re-calcification. These results highlight novel pH regulatory strategies in calcifying cells of a marine species with important implications for our understanding of the mineralization process in times of rapid changes in oceanic pH.


Assuntos
Bicarbonatos/metabolismo , Ouriços-do-Mar/fisiologia , Animais , Transporte Biológico , Calcificação Fisiológica , Carbonato de Cálcio , Concentração de Íons de Hidrogênio , Oceanos e Mares , Prótons , Água do Mar
19.
Water Sci Technol ; 82(4): 651-662, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32970618

RESUMO

The adsorption isotherms of Reactive Red 120 (RR-120) on Brazilian pine-fruit shell activated carbon, at six temperatures (298, 303, 308, 313, 318 and 323 K) and pH = 6, were determined and interpreted using a double layer model with one energy. A statistical physics treatment established the formulation of this model. Steric and energetic parameters related to the adsorption process, such as the number of adsorbed molecules per site, the receptor sites density and the concentration at half-saturation, have been considered. Thermodynamic potential functions such as entropy, internal energy and Gibbs free enthalpy are analyzed, and the choice of the models is based on assumptions in correlation with experimental conditions. By numerical fitting, the investigated parameters were deduced. The theoretical expressions provide a good understanding and interpretation of the adsorption isotherms at the microscopic level. We believe that our work contributes to new theoretical insights on the dye adsorption in order to know the physical nature of the adsorption process.


Assuntos
Carvão Vegetal , Triazinas , Adsorção , Concentração de Íons de Hidrogênio , Cinética , Termodinâmica
20.
Water Sci Technol ; 82(4): 673-682, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32970620

RESUMO

Loess is a typical natural mineral particle distributed widely around the world, and it is inexpensive, readily accessible, and harmless to the environment. In this study, loess was modified by surface grafting copolymerization of functional monomers, such as acrylic acid, N-vinyl pyrrolidone, and N,N-methylenebisacrylamide as a cross-linking agent, which afforded a novel loess-based grafting copolymer (LC-PAVP). After being characterized by scanning electron microscopy, thermal gravimetric analysis and Fourier-transform infrared spectroscopy, its adsorption capacity and mechanism of removing lead ions (Pb2+) were investigated. With the study of the optimal experimental conditions, it was demonstrated that the removal rate of Pb2+ by LC-PAVP can reach up to 99.49% in 60 min at room temperature. It was also found that the kinetic characteristics of the adsorption capacity due to the pseudo-second-order kinetic model and the thermodynamics conformed well with the Freundlich model. In summary, as a lost-cost and eco-friendly loess-based adsorbent, LC-PAVP is a good potential material for wastewater treatment.


Assuntos
Acrilatos , Poluentes Químicos da Água/análise , Adsorção , Concentração de Íons de Hidrogênio , Cinética , Polímeros , Espectroscopia de Infravermelho com Transformada de Fourier , Termodinâmica , Águas Residuárias
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