Time Dependency Experiments Research Articles

From tryptophan-based amides to tertiary amines: Optimization of a butyrylcholinesterase inhibitor series

Lead optimization of a series of tryptophan-based nanomolar butyrylcholinesterase (BChE) inhibitors led to tertiary amines as highly potent, achiral, sp3-rich analogues with better synthetic accessibility and high selectivity over acetylcholinesterase (one to ten thousandfold). Taking it one step further, the introduction of a carbamate warhead on the well-explored reversible scaffold allowed conversion to pseudoirreversible inhibitors that bound covalently to BChE and prolonged the duration of inhibition (half-life of 14.8 h for compound 45a-carbamoylated enzyme). Additionally, N-hydroxyindole was discovered as a novel leaving group chemotype. The covalent mechanism of action was confirmed by time-dependency experiments, progress curve analysis, and indirectly by co-crystallization with the human recombinant enzyme. Two crystal structures of BChE-inhibitor complexes were solved and coupled with the supporting molecular dynamics simulations increased our understanding of the structure-activity relationship, while also providing the necessary structural information for future optimization of this series. Overall, this research demonstates the high versatility and potential of this series of BChE inhibitors.

The Blood-Brain Barrier Permeability of Lignans and Malabaricones from the Seeds of Myristica fragrans in the MDCK-pHaMDR Cell Monolayer Model.

The blood-brain barrier (BBB) permeability of twelve lignans and three phenolic malabaricones from the seeds of Myristica fragrans (nutmeg) were studied with the MDCK-pHaMDR cell monolayer model. The samples were measured by high-performance liquid chromatography and the apparent permeability coefficients (Papp) were calculated. Among the fifteen test compounds, benzonfuran-type, dibenzylbutane-type and arylnaphthalene-type lignans showed poor to moderate permeabilities with Papp values at 10−8–10−6 cm/s; those of 8-O-4′-neolignan and tetrahydrofuran-lignan were at 10−6–10−5 cm/s, meaning that their permeabilities are moderate to high; the permeabilities of malabaricones were poor as their Papp values were at 10−8–10−7 cm/s. To 5-methoxy-dehydrodiisoeugenol (2), erythro-2-(4-allyl-2,6-dimethoxyphenoxy)-1-(3,4-dimethoxyphenyl)-propan-1-ol acetate (6), verrucosin (8), and nectandrin B (9), an efflux way was involved and the main transporter for 6, 8 and 9 was demonstrated to be P-glycoprotein. The time and concentration dependency experiments indicated the main transport mechanism for neolignans dehydrodiisoeugenol (1), myrislignan (7) and 8 was passive diffusion. This study summarized the relationship between the BBB permeability and structure parameters of the test compounds, which could be used to preliminarily predict the transport of a compound through BBB. The results provide a significant molecular basis for better understanding the potential central nervous system effects of nutmeg.

Ethyl Pyruvate Emerges as a Safe and Fast Acting Agent against Trypanosoma brucei by Targeting Pyruvate Kinase Activity.

BackgroundHuman African Trypanosomiasis (HAT) also called sleeping sickness is an infectious disease in humans caused by an extracellular protozoan parasite. The disease, if left untreated, results in 100% mortality. Currently available drugs are full of severe drawbacks and fail to escape the fast development of trypanosoma resistance. Due to similarities in cell metabolism between cancerous tumors and trypanosoma cells, some of the current registered drugs against HAT have also been tested in cancer chemotherapy. Here we demonstrate for the first time that the simple ester, ethyl pyruvate, comprises such properties.ResultsThe current study covers the efficacy and corresponding target evaluation of ethyl pyruvate on T. brucei cell lines using a combination of biochemical techniques including cell proliferation assays, enzyme kinetics, phasecontrast microscopic video imaging and ex vivo toxicity tests. We have shown that ethyl pyruvate effectively kills trypanosomes most probably by net ATP depletion through inhibition of pyruvate kinase (Ki = 3.0±0.29 mM). The potential of ethyl pyruvate as a trypanocidal compound is also strengthened by its fast acting property, killing cells within three hours post exposure. This has been demonstrated using video imaging of live cells as well as concentration and time dependency experiments. Most importantly, ethyl pyruvate produces minimal side effects in human red cells and is known to easily cross the blood-brain-barrier. This makes it a promising candidate for effective treatment of the two clinical stages of sleeping sickness. Trypanosome drug-resistance tests indicate irreversible cell death and a low incidence of resistance development under experimental conditions.ConclusionOur results present ethyl pyruvate as a safe and fast acting trypanocidal compound and show that it inhibits the enzyme pyruvate kinase. Competitive inhibition of this enzyme was found to cause ATP depletion and cell death. Due to its ability to easily cross the blood-brain-barrier, ethyl pyruvate could be considered as new candidate agent to treat the hemolymphatic as well as neurological stages of sleeping sickness.

Efficiency of Low Cost Adsorbents for the Removal of Arsenic from Water

Adsorption is one of the primary processes for removing arsenic from drinking water. This study focuses on developing inexpensive and effective adsorbents to remove arsenic from ground water. Eight different types of adsorbents were prepared. Some of these materials were chemically modified. The efficiency of percentage adsorption of arsenate, As (+III) on different materials were investigated at different pH, contact time and initial concentrations. Out of eight different types of adsorbents, the iron-loaded x ant hated orange waste (Fe-XOW) showed high efficiency for the removal of arsenic. It was found that approximately 83 % of arsenate, As (+III) and 87% of arsenate, As (+V) removal could be achieved at optimum pH of 10 and 4respectively. The significant effect of pH was in the range of 9 to12 for As (+III) and 3 to 5 for As (+V). Time dependency experiments for the arsenite uptake showed that the adsorption rate on Fe-XOW was fast initially for 1 hour, followed by slow attainment of equilibrium at 2.15 hour. Adsorption isotherm test showed that equilibrium adsorption data were better represented by Langmuir model than the Freundlich model and the maximum adsorption (qmax) for As (+III) onto Fe-XOW was found to be 53.47 mg/gm. The concentration of arsenic in water sample was determined by standard silver diethyldithiocarbamate spectrophotometric method (SDDC method).

Biosorption of Ni(II) from aqueous solutions by living and non-living ureolytic mixed culture

The present study explores the ability and the comparison of living and non-living ureolytic mixed culture (UMC) to remove Ni(II) from aqueous solution. Time dependency experiments for the Ni(II) uptake showed that adsorption equilibrium was reached almost 110 and 60 min after addition Ni(II) of 100 mg/L. The kinetic data were analyzed in term of pseudo-first-order and pseudo-second-order expressions. Ni(II) sorption of living UMC was appropriate with pseudo-first-order kinetic ( k 1 = 2.15 h −1, R 2 = 0.93) while non-living UMC sorbed Ni(II) with respect to second-order kinetics ( k 2 = 1.64 g/mg h, R 2 = 0.98). Also, comparison between the biosorption capacity of untreated living and non-living biomass was conducted for removal of Ni(II). The biosorption process was investigated in equilibrium batch tests for Langmiur, Freundlich and Temkin isotherm models. The data pertaining to the sorption dependence upon Ni(II) ion concentration ranged from 5 to 320 mg/L could be fitted to a Freundlich isotherm model. The capacity constants K of Freundlich model for living and non-living UMC were 1.55 and 0.38 mg/g, respectively; the affinity constants 1/ n were 0.47 and 0.75, respectively. Based on the results, the UMC appear to be a potential biosorbent for removal of Ni(II) from wastewater.

Biosorption of lead(II), cadmium(II), copper(II) and nickel(II) by anaerobic granular biomass

Biosorption is potentially an attractive technology for treatment of wastewater for retaining heavy metals from dilute solutions. This study investigated the feasibility of anaerobic granules as a novel type of biosorbent, for lead, copper, cadmium, and nickel removal from aqueous solutions. Anaerobic sludge supplied from a wastewater treatment plant in the province of Quebec was used. Anaerobic granules are microbial aggregates with a strong, compact and porous structure and excellent settling ability. After treatment of the biomass with Ca ions, the cation exchange capacity of the biomass was approximately 111 meq/100g of biomass dry weight which is comparable to the metal binding capacities of commercial ion exchange resins. This work investigated the equilibrium, batch dynamics for the biosorption process. Binding capacity experiments using viable biomass revealed a higher value than those for nonviable biomass. Binding capacity experiments using non-viable biomass treated with Ca revealed a high value of metals uptake. The solution initial pH value affected metal sorption. Over the pH range of 4.0–5.5, pH-related effects were not significant. Meanwhile, at lower pH values the uptake capacity decreased. Time dependency experiments for the metal ions uptake showed that adsorption equilibrium was reached almost 30 min after metal addition. It was found that the q max for Pb 2+, Cd 2+, Cu 2+, and Ni 2+ ions, were 255, 60, 55, and 26 mg/g respectively (1.23, 0.53, 0.87, and 0.44 mmol/g respectively). The data pertaining to the sorption dependence upon metal ion concentration could be fitted to a Langmiur isotherm model. Based on the results, the anaerobic granules treated with Ca appear to be a promising biosorbent for removal of heavy metals from wastewater due to its optimal uptake of heavy metals, its particulate shape, compact porous structure, excellent settling ability, and its high mechanical strength.

Use of chemical modification and spectroscopic techniques to determine the binding and coordination of gadolinium(III) and neodymium(III) ions by alfalfa biomass

Metal pollution in the aqueous environment has become an important issue in the past few decades leading to extensive research in the area of pollution remediation. Most of the recent research in this area has been in bioremediation including phytofiltration and phytoextraction. Although there has been a lot of research done in the field of metal interactions with plants, the actual mechanism(s) and ligands involved are not well understood. Through a series of batch experiments, including pH profiles, time dependency studies, and capacity experiments, we have investigated the binding of Gd(III) and Nd(III) to alfalfa biomass. Batch pH studies showed that the optimum binding was at pH 5.0 for both elements. The time dependency experiments showed that the binding occurs within the first 5 min of contact and remains constant for up to 60 min. In addition, chemical modifications to the alfalfa biomass were performed to indirectly determine the ligands on the biomass responsible for metal binding. For Gd(III) binding, it was shown that the carboxyl groups on the biomass play the most important role in metal ion binding. However, for Nd(III), not only was it found that the carboxyl groups play an important role in the binding, but in addition, the amino groups on the biomass also play an important role in the binding of the metal ions. Further studies using X-ray absorption spectroscopy (XAS) showed that the Gd(III) and Nd(III) ions were bound to the alfalfa biomass through oxygen (or nitrogen ligands), which were coordinated to carbon atoms. The lanthanide complexes within the biomass included some coordinated water molecules.

Investigation of trace level binding of PtCl 6 and PtCl 4 to alfalfa biomass ( Medicago sativa) using Zeeman graphite furnace atomic absorption spectrometry

Batch laboratory experiments were performed to investigate the effects of pH, chemical modification, time dependency, and interference studies on the binding of trace concentrations of hexachloroplatinate(IV) and tetrachloroplatinate(II) to alfalfa biomass. The pH profiles were measured between pH 2.0 and 6.0. It was found that the binding of trace concentrations of platinum(IV and II) to alfalfa biomass was dependent on pH with a maximum binding occurring at pH 3.0 and a minimum at pH 6.0. When the alfalfa biomass was chemically modified (esterified), maximum binding occurred at pH 6.0 for both oxidation states of platinum. From the batch time dependency experiments, it was found that binding took at least 20 min to level off for both platinum oxidation states. Batch experiments were performed with various concentrations of calcium, magnesium, and sodium (0.1, 1.0, 10, 100 and 1000 ppm) and it was found that calcium affected the binding of platinum(II and IV) to the alfalfa biomass. It was determined that magnesium and sodium did not interfere appreciably with the binding of platinum in either of the oxidation states studied. Finally, batch experiments were performed with Mg 2+, Ca 2+ and Na + in solutions at various concentrations, and it was observed that the binding was affected similarly to that by calcium alone.

Use of hop ( Humulus lupulus) agricultural by-products for the reduction of aqueous lead(II) environmental health hazards

The agricultural by-products of the hop plant ( Humulus lupulus L.) were investigated to determine their potential for use in the removal of heavy lead(II) ions from contaminated aqueous solutions. Separate batch laboratory experiments were performed to establish the optimal binding pH, time exposures, and capacity of the metal adsorption for lead(II) ions by dried and ground hop leaves and stems biomass. Results from these studies have shown a pH dependent binding trend from pH 2–6, with optimum binding occurring around pH 5.0. Time dependency experiments showed a rapid adsorption of lead(II) ions within the first 5 min of contact. Binding capacity experiments demonstrated that 74.2 mg of lead(II) were bound per gram of leaf biomass. Similarly overall capacity was seen for the leaves and stems. Desorption of 99% of the bound lead(II) ions was achieved by exposing the metal laden biomass to 0.5 M sodium citrate. Further experiments were performed with silica-immobilized hop tissues to determine the lead(II) binding ability under flow conditions. Comparison studies were performed with ion-exchange resins to evaluate the binding ability and to gain further insight into the metal binding mechanism. X-ray absorption spectroscopy experiments were also utilized to gain further insight into the possible lead(II) binding mechanism by the hop plant tissue. Results from these studies indicate that carboxyl ligands are involved in the binding of lead(II) from aqueous solution. These findings show that the use of hop agricultural waste products may be a viable alternative, for the removal and recovery of aqueous lead(II) ions from contaminated waters.

Bovine interferon-tau stimulates the Janus kinase-signal transducer and activator of transcription pathway in bovine endometrial epithelial cells.

Trophoblastic bovine interferon-tau (bIFN-tau) suppresses luteolytic pulses of endometrial prostaglandin F(2alpha) (PGF(2alpha)) at the time of maternal recognition of pregnancy. This results in maintenance of the corpus luteum in cattle. The hypothesis that effects of bIFN-tau in the endometrium were through activation of the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway of signal transduction was tested. Whole cell, cytosolic, and nuclear extracts from bovine endometrial cells treated with bIFN-tau were analyzed by immunoprecipitation, immunoblotting, and electrophoretic mobility shift assays in a series of dose- and time-dependency experiments. Bovine IFN-tau stimulated tyrosine phosphorylation, homo- and heterodimer formation, nuclear translocation, and DNA binding of STAT proteins 1, 2, and 3. Moreover, bIFN-tau induced synthesis of interferon-regulatory factor. In conclusion, bIFN-tau stimulates the JAK-STAT pathway in the bovine endometrium. It is proposed that activation of the JAK-STAT pathway is involved in regulating the antiluteolytic effects of bIFN-tau.

Biosorption of actinides from dilute waste actinide solution by egg-shell membrane.

Removal of radioactive elements from the effluent and waste aqueous solutions is an important problem. In previous laboratory batch experiments, hen egg-shell membrane (ESM) was stable as an insoluble protein and was very capable of binding heavy metal ions from aqueous solution. Batch laboratory pH profile, time dependency, and capacity experiments were performed to determine the binding of uranium (U) and thorium (Th) to ESM. Batch pH profile experiments indicated that the optimum pH for binding these actinides was approx 6.0 (U) or 3.0 (Th). The adsorption isotherms were developed at pH 5.0 (U) or 3.0 (Th) at 25 degrees C, and the adsorption equilibrium data fitted both Langmuir and Freundlich models. The maximum uptakes by the Langmuir model were about 240 mg U/g and 60 mg Th/g dry weight ESM. In addition, their adsorption capacities increased as salt concentration increased. ESM could also accumulate uranium from dilute aqueous solution by adjusting to the optimum pH. These results showed that ESM was effective for removing actinides from solution and would be useful in filtration technology to remove actinides from aqueous solution.

Potential Use of Bagasse Pith for the Treatment of Wastewater Containing Metals

Bagasse pith was tested for its ability to remove cadmium and lead ions from aqueous solutions under two modes of operation, i.e. in batch systems and continuous flow systems. Time dependency experiments showed a very rapid adsorption of these cations by bagasse pith. Thus, 62% of cadmium ions and 84% of lead ions were taken up by the bagasse pith at pH 6 and ca. 99% of each cation was recovered by treatment of the pith with 0.1 M HCl. The breakthrough capacity, Q0.5, the number of theoretical plates and the theoretical plate height were calculated. The capacity of bagasse pith was found to be 6.2 and 8.5 mequiv./g for cadmium and lead ions, respectively. The feasibility of using bagasse pith for cadmium and lead ion removal from wastewater streams was addressed.

Phytofiltration of hazardous cadmium, chromium, lead and zinc ions by biomass of Medicago sativa (Alfalfa)

Previous laboratory batch experiments of Medicago sativa (Alfalfa) indicated that the African shoots population had an appreciable ability to bind copper(II) and nickel(II) ions from aqueous solution. Batch laboratory pH profile, time dependency and capacity experiments were performed to determine the binding ability of the African shoots for cadmium(II), chromium(III), chromium(VI), lead(II), and zinc(II). Batch pH profile experiments for the mentioned ions indicated that the optimum pH for metal binding is approximately 5.0. Time dependency experiments for all the metals studied showed that metal binding to the African alfalfa shoots occurred within 5 min. Binding capacity experiments revealed the following amounts of metal ions bound per gram of biomass: 7.1 mg Cd(II), 7.7 mg Cr(III), 43 mg Pb(II), and 4.9 mg Zn(II). However, no binding occurred for chromium(VI). Nearly all of the metals studied were recoverable by treatment with 0.1 M HCl. Column experiments were performed to study the binding of Cd(II), Cr(III), Cr(VI), Pb(II) and Zn(II) to silica-immobilized African alfalfa shoots under flow conditions. These experiments showed that the silica immobilized African alfalfa shoots were effective for removing metal ions from solution, and over 90% of the bound Pb(II), Cu(II), Ni(II), and Zn(II), and over 70%Cd(II), were recovered after treatment with 10 bed volumes of 0.1 M HCl. The results from these studies will be useful for a novel phytofiltration technology to remove and recover heavy metal ions from aqueous solution.

UPTAKE OF COPPER IONS FROM SOLUTION BY DIFFERENT POPULATIONS OF MEDICAGO SATIVA (ALFALFA)

ABSTRACT Seven different varieties of alfalfa plant tissues were tested for their ability to uptake copper ions from solution. Laboratory experiments were performed to determine the pH profiles, time dependency, capacity of copper uptake and desorption of the metal bound. Also, optimal biomass drying conditions (oven dried and lyophilized) as well as buffering effects on the copper uptake were examined. Differences in the roots and shoots were examined to determine the metal binding properties of seven alfalfa populations. Protein and crude fiber content was determined for all biomasses studied. Most of the biomasses studied showed a high affinity for copper ions as the pH increased from 2 to 6 with optimum copper binding around pH 5.0. Time dependency experiments for copper binding showed a very rapid adsorption of the copper ions by the various alfalfa species. Binding capacity experiments for copper binding showed that the capacities of the shoots are very similar, with African and Ladak germplasms bei...