Metabolic disorders, including hyperglycemia, characterize type-2 diabetes. One of the treatment methods used for postprandial hyperglycemia includes using potential therapeutic agents to inhibit ...α-amylase activity. This study utilized fractional design and the simplex method to optimize in vitro microscale assay inhibition conditions using Miller’s reaction. In addition, the effect of substrate concentration on enzyme activity was analyzed. Enzyme concentration of 0.15 U mL-1 and pre- and post-incubation times of 7.2 and 5.5 min, respectively, in water bath (15.6 min) equipment, were set up for optimized condition for the enzyme activity. Analytical validation was performed based on different international guidelines. Km was found to be 0.38 mg mL-1. Linearity was obtained at the acarbose concentration of 1.5 µg mL-1 and 5 µg mL-1. The IC50 for the positive control was found to be 0.6 µg mL-1. The relative standard deviation and Z value were found to be <4% and >0.93, respectively. Additionally, the optimized assay was applied to extracts from five different plants. Two plant extracts (Zanthoxylum fagara and Chrysactinia mexicana) inhibited α-amylase activity. The optimized and validated method was accurate, precise, and linear
The quantification of low-abundance secondary metabolites in plant extracts is an analytical problem that can be addressed by different analytical platforms, the most common being those based on ...chromatographic methods coupled to a high-sensitivity detection system. However, in recent years nuclear magnetic resonance (NMR) has become an analytical tool of primary choice for this type of problem because of its reliability, inherent simplicity in sample preparation, reduced analysis time, and low solvent consumption. The versatility of strategies based on quantitative NMR (qNMR), such as internal and external standards and electronic references, among others, and the need to develop validated analytical methods make it essential to compare procedures that must rigorously satisfy the analytical well-established acceptance criteria for method validation. In this work, two qNMR methods were developed for the quantification of hepatodamianol, a bioactive component of T. diffusa. The first method was based on a conventional external standard calibration, and the second one was based on the pulse length-based concentration determination (PULCON) method using the ERETIC2 module as a quantitation tool available in TopSpin software. The results show that both procedures allow the content of the analyte of interest in a complex matrix to be determined in a satisfactory way, under strict analytical criteria. In addition, ERETIC2 offers additional advantages such as a reduction in experimental time, reagent consumption, and waste generated.
One primary drawback of enzyme catalysis at industrial scale is the short-term service life of the enzymes, they lose their activity due to oxidation or other processes which results in less ...stability and a shorter lifetime thereby rendering them less efficient. An effective way to increase the stability of the enzymes is to attach them to nanoparticles. In this work, the polymer Eudragit® L 100-55 sensitive to pH was used to prepare laccase polymeric nanoparticles by the double-emulsion solvent evaporation approach. The size and morphology of the nanoparticles obtained were evaluated—as well as the encapsulation efficiency and zeta potential. pH effect on activity and stability was compared between free and immobilized laccase. Their stability was also determined in a sequential assay involving acidic pHs up to alkaline ones. The nanoparticles had a spherical shape with a mean size of 147 nm, zeta potential of −22.7 mV at pH 7.0 and load efficiency of 87%. The optimum pH of both free and immobilized laccases was 3.0, being the nanoparticles more stable at acidic pHs. Thus, this would be the first report of obtaining laccase nanoparticles with potential application in animal feed due to the stability conferred to enzymatic activity at pHs similar to those present in the gastrointestinal tract of rabbits, which would allow their potential use in animal feed.
The usefulness of traditional plants in Mexico to treat human ailments has been known since ancient times. This work evaluated the antimicrobial, anticoagulant, antioxidant, cytotoxic, and ...anti-inflammatory potential of ethanolic extracts of Aloe vera, Equisetum arvense, Mimosa tenuiflora, Lippia graveolens, and Syzygium aromaticum. The antimicrobial activity of the extracts was evaluated against Streptococcus mutans and Streptococcus sorbinus; a significant inhibitory effect of the L. graveolens extract on both bacteria was observed at concentration levels of 250 µg/mL and greater. The anticoagulant activity was evaluated in terms of prothrombin time (PT) and activated partial thromboplastin time (APTT), A. vera and M. tenuiflora extracts showed no significant difference (p ˂ 0.05) in PT compared with the control, and for APTT the extracts of A. vera, L. graveolens, and S. aromaticum decreased the APTT significantly (p ˂ 0.05) compared with the control. The antioxidant potential by DPPH assay indicated that the E. arvense extract behaved statistically the same as the control. The cytotoxic activity was evaluated in HGF-1 cells using the fluorometric microculture cytotoxicity assay technique, and none of the extracts was toxic at 125 and 250 µg/mL concentrations. Finally, the anti-inflammatory activity was evaluated using ELISA, where the A. vera extract showed the best anti-inflammatory capacity. Further research on the search for bioactive metabolites and elucidation of action mechanisms of the most promising extracts will be carried out.
Autoimmune lymphoproliferative syndrome (ALPS) is a rare disease defined as a defect in the lymphocyte apoptotic pathway. Currently, the diagnosis of ALPS is based on clinical aspects, defective ...lymphocyte apoptosis and mutations in
,
and
genes. Despite this, ALPS has been misdiagnosed. The aim of this work was to go one step further in the knowledge of the disease, through a molecular and proteomic analysis of peripheral blood mononuclear cells (PBMCs) from two children, a 13-year-old girl and a 6-year-old boy, called patient 1 and patient 2, respectively, with clinical data supporting the diagnosis of ALPS.
,
and
genes from both patients were sequenced, and a sample of the total proteins from patient 1 was analyzed by label-free proteomics. Pathway analysis of deregulated proteins from PBMCs was performed on the STRING and PANTHER bioinformatics databases. A mutation resulting in an in-frame premature stop codon and protein truncation was detected in the
gene from patient 2. From patient 1, the proteomic analysis showed differences in the level of expression of proteins involved in, among other processes, cell cycle, regulation of cell cycle arrest and immune response. Noticeably, the most down-regulated protein is an important regulator of the cell cycle process. This could be an explanation of the disease in patient 1.
Recently, pharmaceutical and personal care products (PPCPs) have received considerable attention because of their increasing use. Analysis of PPCPs presents a significant analytical challenge, with ...high-performance liquid chromatography (HPLC) in reversed-phase mode, as the most widely used analytical technique. To facilitate the optimization of the procedures that are applied in the early stages of sample preparation, a simple and fast HPLC method is proposed in this work for the separation of some PPCPs with a wide range of hydrophilicity. Two columns were evaluated (Atlantis dC18 and Discovery HS F5); as for mobile phases: a formate buffer (40 mmol L
, pH 4) and methanol were tested in a gradient mode. The fluorinated column allowed better separation in a shorter time and better resolution for all analytes (
> 1). The proposed method delivered good performance for the tracing of PPCPs and is a suitable alternative to traditional C18-based HPLC methods.
Eudragit® polymers have proven their potential as a means to control the release of aqueous insoluble drugs in various delivery systems as polymer nanoparticles (PNPs). The size (S) and ...polydispersity index (PDI) of PNPs are crucial factors for their interaction with biological systems from a pharmaceutical standpoint. This study aimed to determine the impact of the volumes of the organic phase (OP) and aqueous phase (AP), as well as the polymer amount (PA), on the size and PDI of PNPs prepared using the nanoprecipitation method for encapsulating quercetin (Qr). The study also evaluated the toxic effects of PNPs on human erythrocytes. The PNPs were prepared using preformed polymers derived from methacrylic acid and polyvinyl alcohol (PVA) as a surfactant. The nanoprecipitation technique enabled the production of particles smaller than 200 nm with a PDI lower than 0.2, and the study established the significant impact (p < 0.05) of the three variables related to the polymers and solvents. The selected PNPs contained 5 mg of Qr and 50 mg of Eudragit polymers (1:10 w/w Eudragit® EPO, E100, L100, and Eudragit L100-55) and diverse concentrations of PVA. The study found that including PVA in the AP increased the Qr encapsulation by up to 98%. The hemolytic potential of Eudragit® PNPs and Qr was assessed in human erythrocytes, with no significant cytotoxic activity observed (p < 0.001) compared with the control. In conclusion, via the nanoprecipitation technique, preparing PNPs with defined and homogeneous S to entrap the flavonol Qr efficiently is possible.
Solid-phase microextraction (SPME) is a sample preparation technique with many applications that is being continuously developed. In this technique, the type of fiber coating plays a crucial role for ...extraction efficiency. Currently available commercial coatings have certain drawbacks that have been overcome by the development of new coatings based on novel materials; these have improved the efficiency of extraction, selectivity and stability of commercial coatings. Pharmaceutical and personal care products (PPCPs) are one of the most important groups of emerging contaminants; however, some studies suggest that these compounds can cause adverse health effects. No official monitoring protocols for these compounds are currently available, so the establishment of analytical methods that allow their determination in environmental samples is required. The complexity of environmental samples together with the low concentration levels of these compounds makes necessary the use of sample preparation techniques capable of removing interferences, as well as preconcentrated analytes, and SPME is a very promising alternative to achieve this. This review describes the recent developments in SPME with classical and novel coatings and its applications for PPCP determination in environmental samples.
Strongyloidiasis is a parasitosis that represents a public health problem, in tropical regions. The present study aimed to investigate the anthelmintic effects of several extracts of Argemone ...mexicana, as well as its main component berberine (Ber) against the third-stage larvae (L3) of Strongyloides venezuelensis in-vitro experiments. Also, the anti-hemolytic activity of the extract, fractions, and Ber were tested in human erythrocytes. A dose-response anthelminthic bioassay demonstrated Ber as the most effective component, followed by methanolic subfraction (Fr3) and finally the crude extract of A. mexicana (Am) showing LC50 response values of 1.6, 19.5, and 92.1 μg/mL, at 96 h respectively. Also, Am, Fr3, and Ber did not produce significant hemolysis against human erythrocytes (p ≤ 0.05). Am and Fr3 showed erythrocyte protection effect capacity at the membrane level (p ≤ 0.05). Furthermore, Ber was found to have an antioxidant activity of 168.18 μg/mL. According to the results, the Fr3 of A. mexicana, and particularly Ber, exhibited potent in-vitro effects against L3 of S. venezuelensis, without hemolytic activity against human erythrocytes and presented good antioxidant capacity. In conclusion, the extracts of A. mexicana and the main component have activity against S. venezuelensis, nevertheless, further studies are required to elucidate the mechanism of action.
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•Anthelmintic activity of A. mexicana methanolic extract, active fraction, and it ‘s main component.•Development of a new alternative to a conventional anthelmintics.•The capacity of the berberine alkaloid is reported, which has a response against the viability of S. venezuelensis (L3).•The safety of the methanolic extract, active fraction, and berberine, was evaluated by an in-vitro hemolytic bioassay.
Parasitic infections represent one of the main public health problems in humans according to the WHO. Therefore, the need has arisen to find new treatments that can be used as an alternative cure to ...parasitosis. We aimed to investigate the in-vitro effects of the methanolic extract of
as well as its main component, quercetin against
and
.
For this purpose, the in-vitro activity of the methanol extract of
also its main component, quercetin, against trophozoites of
and
was evaluated, using the microassay technique. Furthermore, the antioxidant activity was determined. Finally, the cytotoxic and cytoprotective capacity was determined using the hemolysis technique.
The IC
indicated that quercetin significantly (
< 0.05) inhibited the growth rate of the trophozoite stage of
and
in comparison to the methanolic extract of
(KalL). Also, quercetin significantly (
< 0.05) was a better antioxidant as compared with the positive control. In the evaluation of cytotoxicity effects, it could be observed that KalL as compared with quercetin exhibited more cytotoxicity against human erythrocytes. Quercetin significantly (
< 0.001) exhibited better cytoprotective activity compared to KalL.
Both
methanolic extract and quercetin alone demonstrated high antiparasitic activity against
and
. However, the in-vivo efficacy of
and quercetin also requires to be evaluated using an animal model.
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