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1.
Pharmaceutics ; 16(5)2024 May 16.
Artículo en Inglés | MEDLINE | ID: mdl-38794330

RESUMEN

Biological nanoparticles (NPs), such as extracellular vesicles (EVs), exosome-mimetic nanovesicles (EMNVs) and nanoghosts (NGs), are perspective non-viral delivery vehicles for all types of therapeutic cargo. Biological NPs are renowned for their exceptional biocompatibility and safety, alongside their ease of functionalization, but a significant challenge arises when attempting to load therapeutic payloads, such as nucleic acids (NAs). One effective strategy involves fusing biological NPs with liposomes loaded with NAs, resulting in hybrid carriers that offer the benefits of both biological NPs and the capacity for high cargo loads. Despite their unique parameters, one of the major issues of virtually any nanoformulation is the ability to escape degradation in the compartment of endosomes and lysosomes which determines the overall efficiency of nanotherapeutics. In this study, we fabricated all major types of biological and hybrid NPs and studied their response to the acidic environment observed in the endolysosomal compartment. In this study, we show that EMNVs display increased protonation and swelling relative to EVs and NGs in an acidic environment. Furthermore, the hybrid NPs exhibit an even greater response compared to EMNVs. Short-term incubation of EMNVs in acidic pH corresponding to late endosomes and lysosomes again induces protonation and swelling, whereas hybrid NPs are ruptured, resulting in the decline in their quantities. Our findings demonstrate that in an acidic environment, there is enhanced rupture and release of vesicular cargo observed in hybrid EMNVs that are fused with liposomes compared to EMNVs alone. This was confirmed through PAGE electrophoresis analysis of mCherry protein loaded into nanoparticles. In vitro analysis of NPs colocalization with lysosomes in HepG2 cells demonstrated that EMNVs mostly avoid the endolysosomal compartment, whereas hybrid NPs escape it over time. To conclude, (1) hybrid biological NPs fused with liposomes appear more efficient in the endolysosomal escape via the mechanism of proton sponge-associated scavenging of protons by NPs, influx of counterions and water, and rupture of endo/lysosomes, but (2) EMNVs are much more efficient than hybrid NPs in actually avoiding the endolysosomal compartment in human cells. These results reveal biochemical differences across four major types of biological and hybrid NPs and indicate that EMNVs are more efficient in escaping or avoiding the endolysosomal compartment.

2.
Curr Rheumatol Rev ; 2024 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-38415452

RESUMEN

BACKGROUND: Ankylosing spondylitis (AS) is a chronic inflammatory disease known for causing pain, stiffness, and reduced mobility in the axial skeleton. Adalimumab, a tumor necrosis factor (TNF) inhibitor, has emerged as a promising therapeutic option for AS. METHODS: This systematic review involved a comprehensive search of randomized controlled trials related to AS treatment, conducted in major databases such as MEDLINE, Google Scholar, and PubMed. The search terms encompassed ankylosing spondylitis, adalimumab, methotrexate, other non-biologic DMARDs, glucocorticoids, NSAIDs, and analgesics. A total of 14 randomized controlled trials with 4,500 participants were included in the review. RESULTS: The review's results revealed that adalimumab demonstrated notable superiority when compared to a placebo. It effectively reduced disease activity, improved physical function, and lowered inflammatory markers such as C-reactive protein and erythrocyte sedimentation rate. Adalimumab demonstrated a favorable safety profile, with adverse events comparable to those observed with placebo. CONCLUSION: Based on the results, adalimumab is deemed an effective treatment for AS, showcasing its potential as a first-line therapeutic option. Notably, no significant increase in adverse events was observed compared to placebo. However, the conclusion emphasizes the need for further studies with extended follow-up durations to ascertain the long-term efficacy and safety of adalimumab in AS management. This systematic review provides valuable insights supporting the use of adalimumab in the treatment of AS and underscores the importance of ongoing investigations into its long-term effects to optimize its clinical utilization in AS patients.

3.
J Pharm Biomed Anal ; 239: 115912, 2024 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-38128161

RESUMEN

Olive trees are one of the most widely cultivated fruit trees in the world. The chemical compositions and biological activities of olive tree fruit and leaves have been extensively researched for their nutritional and health-promoting properties. In contrast, limited data have been reported on olive flowers. The present study aimed to analyse bioactive compounds in olive flower extracts and the effect of fermentation-assisted extraction on phenolic content and antioxidant activity. High-performance thin-layer chromatography (HPTLC) hyphenated with the bioassay-guided detection and spectroscopic identification of bioactive compounds was used for the analysis. Enzymatic and bacterial in situ bioassays were used to detect COX-1 enzyme inhibition and antibacterial activity. Multiple zones of antibacterial activity and one zone of COX-1 inhibition were detected in both, non-fermented and fermented, extracts. A newly developed HPTLC-based experimental protocol was used to measure the high-maximal inhibitory concentrations (IC50) for the assessment of the relative potency of the extracts in inhibiting COX-1 enzyme and antibacterial activity. Strong antibacterial activities detected in zones 4 and 7 were significantly higher in comparison to ampicillin, as confirmed by low IC50 values (IC50 = 57-58 µg in zone 4 and IC50 = 157-167 µg in zone 7) compared to the ampicillin IC50 value (IC50 = 495 µg). The COX-1 inhibition by the extract (IC50 = 76-98 µg) was also strong compared to that of salicylic acid (IC50 = 557 µg). By comparing the locations of the bands to coeluted standards, compounds from detected bioactive bands were tentatively identified. The eluates from bioactive HPTLC zones were further analysed by FTIR NMR, and LC-MS spectroscopy. Multiple zones of antibacterial activity were associated with the presence of triterpenoid acids, while COX-1 inhibition was related to the presence of long-chain fatty acids.


Asunto(s)
Olea , Olea/química , Cromatografía en Capa Delgada/métodos , Árboles , Extractos Vegetales/química , Flores/química , Antibacterianos/farmacología , Antibacterianos/análisis , Antioxidantes/farmacología , Espectroscopía de Resonancia Magnética , Ampicilina/análisis , Bioensayo/métodos
4.
Molecules ; 28(19)2023 Sep 29.
Artículo en Inglés | MEDLINE | ID: mdl-37836723

RESUMEN

Solid-phase microextraction (SPME) is a sample preparation technique which utilizes small amounts of an extraction phase for the extraction of target analytes from investigated sample matrices. Its simplicity of use, relatively short sample processing time, and fiber reusability have made SPME an attractive choice for many analytical applications. SPME has been widely applied to the sampling and analysis of environmental, food, aromatic, metallic, forensic, and pharmaceutical samples. Solid phase microextraction is used in horticultural crops, for example, to determine water and soil contaminants (pesticides, alcohols, phenols, amines, herbicides, etc.). SPME is also used in the food industry to separate biologically active substances in food products for various purposes, for example, disease prevention, determining the smell of food products, and analyzing tastes. SPME has been applied to forensic analysis to determine the alcohol concentration in blood and that of sugar in urine. This method has also been widely used in pharmaceutical analysis. It is a solvent-free sample preparation technique that integrates sampling, isolation, and concentration. This review focuses on recent work on the use of SPME techniques in the analysis of food and horticultural crops.


Asunto(s)
Productos Agrícolas , Microextracción en Fase Sólida , Microextracción en Fase Sólida/métodos , Solventes , Etanol , Preparaciones Farmacéuticas
5.
J Chromatogr A ; 1706: 464241, 2023 Sep 13.
Artículo en Inglés | MEDLINE | ID: mdl-37541060

RESUMEN

This study compares different solvent systems with the use of spontaneous fermentation on the phytochemical composition of leaf extracts from a locally grown white variety of common fig (Ficus carica Linn.). The aim was to detect and identify bioactive compounds that are responsible for acetylcholinesterase (AChE), α-amylase and cyclooxygenase-1 (COX-1) enzyme inhibition, and compounds that exhibit antimicrobial activity. Bioactive zones in chromatograms were detected by combining High-performance thin-layer chromatography (HPTLC) with enzymatic and biological assays. A new experimental protocol for measuring the relative half-maximum inhibitory concentration (IC50) was designed to evaluate the potency of the extracts compared to the potency of known inhibitors. Although the IC50 of the fig leaf extract for α-amylase and AChE inhibition were significantly higher when compared to IC50 for acarbose and donepezil, the COX-1 inhibition by the extract (IC50 = 627 µg) was comparable to that of salicylic acid (IC50 = 557 µg), and antimicrobial activity of the extract (IC50 = 375-511 µg) was similar to ampicillin (IC50 = 495 µg). Four chromatographic zones exhibited bioactivity. Compounds from detected bioactive bands were provisionally identified by comparing the band positions to coeluted standards, and by Fourier transform infrared (FTIR) spectra from eluted zones. Flash chromatography was used to separate selected extract into fractions and isolate fractions that are rich in bioactive compounds for further characterisation with nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography-mass spectrometry (LC-MS) analysis. The main constituents identified were umbelliferon (zone 1), furocoumarins psoralen and bergapten (zone 2), different fatty acids (zone 3 and 4), and pentacyclic triterpenoids (calotropenyl acetate or lupeol) and stigmasterol (zone 4).


Asunto(s)
Antiinfecciosos , Ficus , Cromatografía en Capa Delgada , Ficus/química , Extractos Vegetales/farmacología , Extractos Vegetales/química , Acetilcolinesterasa , alfa-Amilasas , Triterpenos Pentacíclicos , Antiinfecciosos/farmacología
6.
J Pharm Biomed Anal ; 227: 115308, 2023 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-36827737

RESUMEN

Extracts of two Salvia species, Salvia apiana (white sage) and Salvia officinalis (common sage) were screened for phytoconstituents with the ability to act as antidiabetic, cognitive enhancing, or antimicrobial agents, by hyphenation of high-performance thin-layer chromatography with enzymatic and microbial effect directed assays. Two bioactive zones with α-amylase inhibition (zone 1 and zone 2), 3 zones for acetylcholinesterase inhibition (zones 3, 4 and 5), and two zones for antimicrobial activity (zones 4 and 5) were detected. The compounds from the five bioactive zones were initially identified by coelution with standards and comparing the RF values of standards to the bioautograms. Identity was confirmed with ATR-FTIR spectra of the isolated compounds from the bioactive zones. A significantly higher α-amylase and acetylcholinesterase inhibition of S. apiana leaf extract was associated with a higher flavonoid and diterpenoid content. Fermented S. officinalis extract exhibited a significantly higher ability to inhibit α-amylase compared to other non-fermented extracts from this species, due to increased extraction of flavonoids. The ATR-FTIR spectra of 2 zones with α-amylase inhibition, indicated that flavonoids and phenolic acids were responsible for α-amylase inhibition. Multiple zones of acetylcholinesterase inhibition were related to the presence of phenolic abietane diterpenoids and triterpenoid acids. The presence of abietane diterpenoids and triterpenoid acids was also found responsible for the mild antimicrobial activity. Flash chromatography was used to isolate sufficient amounts of bioactive compounds for further characterisation via NMR and MS spectroscopy. Five compounds were assigned to the zones where bioactivity was observed: cirsimaritin (zone 1), a caffeic acid polymer (zone 2), 16-hydroxyrosmanol (zone 3), 16-hydroxycarnosic acid (zone 4), oleanolic and ursolic acids (zone 5).


Asunto(s)
Antiinfecciosos , Salvia , Triterpenos , Salvia/química , Acetilcolinesterasa , Cromatografía en Capa Delgada/métodos , Abietanos , Antiinfecciosos/farmacología , Flavonoides , alfa-Amilasas , Extractos Vegetales/química , Antioxidantes/farmacología
7.
Biomedicines ; 10(11)2022 Oct 29.
Artículo en Inglés | MEDLINE | ID: mdl-36359270

RESUMEN

As the number of therapeutic protein products is growing rapidly, there is a strong need for the development of bioanalytical methods that are easy to perform, specific, sensitive, robust, and affordable. Methods for immunogenicity evaluation of therapeutic proteins take an important place in this field of bioanalytics. The aim of the study was to develop a method for immunogenicity testing of the novel RPH-104 drug using the Affinity Capture Elution (ACE) ELISA technique. RPH-104 is a promising Interleukin-1 (IL-1) inhibitor that is currently undergoing a series of clinical studies, including those on socially significant and orphan diseases. The developed method was validated for assay cut-point, sensitivity, selectivity, drug tolerance, hook effect, specificity, precision, and stability. Method sensitivity was established at 114.9 ng/mL, while low and high positive controls were equal to anti-RPH-104 antibody concentrations of 155 ng/mL and 2500 ng/mL, respectively. Method specificity was confirmed in the presence of the interfering compounds, namely IL-1α, IL-1ß, and IL1-Ra. The developed and validated ELISA method was successfully applied to subject samples.

8.
J Pharm Biomed Anal ; 219: 114916, 2022 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-35809514

RESUMEN

In this study, effect-directed analysis (EDA) (i.e. TLC hyphenated with an in situ MTT (3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide) antimicrobial assay), was used for screening and identification of antimicrobials in olive leaf extract. EDA detected that the same compounds exhibited significant antimicrobial activity against bacterial species of the genera Enterococcus (E. faecalis), Escherichia (E. coli), Streptococcus (S. mutans) and Staphylococcus (S. aureus). Flash chromatography-fractionation was used to isolate antimicrobial compounds in olive leaf extract. The active compounds were identified as maslinic acid and oleanolic acid by comparing RF values of the detected active bands with the standard reference materials, with identity confirmed with NMR and ATR-FTIR spectroscopy. Maslinic and oleanolic acids were tested on the E. faecalis strain (which displayed the highest sensitivity in the MTT assay) to determine their inhibiting concentration 50% (IC50) and minimum bactericidal concentrations.


Asunto(s)
Antiinfecciosos , Ácido Oleanólico , Antibacterianos/química , Antibacterianos/farmacología , Cromatografía en Capa Delgada , Escherichia coli , Pruebas de Sensibilidad Microbiana , Olea , Ácido Oleanólico/química , Extractos Vegetales/química , Staphylococcus aureus
9.
Molecules ; 26(22)2021 Nov 16.
Artículo en Inglés | MEDLINE | ID: mdl-34833984

RESUMEN

The aim of this study was to analyse the effect of spontaneous microbial maceration on the release and extraction of the flavonoids and phenolics from olive leaves. Bioprofiling based on thin-layer chromatography effect-directed detection followed by ATR-FTIR spectroscopy proved to be a reliable and convenient method for simultaneous comparison of the extracts. Results show that fermentation significantly enhances the extraction of phenolic compounds and flavonoids. The polyphenolic content was increased from 6.7 µg GAE (gallic acid equivalents) to 25.5 µg GAE, antioxidants from 10.3 µg GAE to 25.3 µg GAE, and flavonoid content from 42 µg RE (rutin equivalents) to 238 µg RE per 20 µL of extract. Increased antioxidant activity of fermented ethyl acetate extracts was attributed to the higher concentration of extracted flavonoids and phenolic terpenoids, while increased antioxidant activity in fermented ethanol extract was due to increased extraction of flavonoids as extraction of phenolic compounds was not improved. Lactic acid that is released during fermentation and glycine present in the olive leaves form a natural deep eutectic solvent (NADES) with significantly increased solubility for flavonoids.


Asunto(s)
Flavonoides/análisis , Olea/química , Extractos Vegetales/química , Hojas de la Planta/química , Polifenoles/análisis , Espectroscopía Infrarroja por Transformada de Fourier
10.
Viruses ; 13(11)2021 11 11.
Artículo en Inglés | MEDLINE | ID: mdl-34835064

RESUMEN

CRISPR/Cas is a powerful tool for studying the role of genes in viral infections. The invention of CRISPR screening technologies has made it possible to untangle complex interactions between the host and viral agents. Moreover, whole-genome and pathway-specific CRISPR screens have facilitated identification of novel drug candidates for treating viral infections. In this review, we highlight recent developments in the fields of CRISPR/Cas with a focus on the use of CRISPR screens for studying viral infections and identifying new candidate genes to aid development of antivirals.


Asunto(s)
Sistemas CRISPR-Cas , Técnicas Genéticas , Estudio de Asociación del Genoma Completo/métodos , Ensayos Analíticos de Alto Rendimiento/métodos , Virosis/genética , Virosis/virología , Virus/genética , Descubrimiento de Drogas , Interacciones Microbiota-Huesped , Humanos
11.
Molecules ; 26(19)2021 Oct 07.
Artículo en Inglés | MEDLINE | ID: mdl-34641608

RESUMEN

The effect of spontaneous fermentation by lactic acid bacteria on the extraction yield of bioactive compounds and antioxidant activity from rosemary leaf extracts was investigated using high-performance thin-layer chromatography (HPTLC). Brining and spontaneous fermentation with lactic acid bacteria more than doubled extraction of polyphenolics and antioxidants from the rosemary leaves. The results show that lactic acid fermentation enhances antioxidant activity in extracts by increasing the total phenolic content but does not increase extraction of phytosterols. Increased extraction of phenolic oxidants during fermentation assisted extraction, results from the in situ generated natural eutectic solvent from the plant sample. ATR-FTIR spectra from the bioactive bands suggests that this increased antioxidant activity is associated with increased extraction of rosmarinic acid, depolymerised lignin, abietane diterpenoids and 15-hydroxy-7-oxodehydroabietic acid.


Asunto(s)
Antioxidantes/química , Antioxidantes/metabolismo , Lactobacillales/metabolismo , Extractos Vegetales/química , Extractos Vegetales/metabolismo , Rosmarinus/química , Rosmarinus/metabolismo , Abietanos/química , Abietanos/metabolismo , Cromatografía en Capa Delgada , Cinamatos/química , Cinamatos/metabolismo , Depsidos/química , Depsidos/metabolismo , Fermentación , Humanos , Lignina/química , Lignina/metabolismo , Fenoles/química , Fenoles/metabolismo , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Espectroscopía Infrarroja por Transformada de Fourier , Ácido Rosmarínico
12.
Viruses ; 13(7)2021 07 15.
Artículo en Inglés | MEDLINE | ID: mdl-34372578

RESUMEN

Viral infections cause a variety of acute and chronic human diseases, sometimes resulting in small local outbreaks, or in some cases spreading across the globe and leading to global pandemics. Understanding and exploiting virus-host interactions is instrumental for identifying host factors involved in viral replication, developing effective antiviral agents, and mitigating the severity of virus-borne infectious diseases. The diversity of CRISPR systems and CRISPR-based tools enables the specific modulation of innate immune responses and has contributed impressively to the fields of virology and immunology in a very short time. In this review, we describe the most recent advances in the use of CRISPR systems for basic and translational studies of virus-host interactions.


Asunto(s)
Antivirales/inmunología , Antivirales/farmacología , Sistemas CRISPR-Cas , Virosis/inmunología , Animales , Exorribonucleasas/metabolismo , Interacciones Microbiota-Huesped/inmunología , Humanos , Evasión Inmune , Inmunidad Innata , Interferones/genética , Interferones/inmunología , Edición de ARN , Transcriptoma , Virosis/virología , Internalización del Virus , Replicación Viral/efectos de los fármacos
13.
J Chromatogr A ; 1647: 462153, 2021 Jun 21.
Artículo en Inglés | MEDLINE | ID: mdl-33957349

RESUMEN

Pittosporum angustifolium, known as gumbi gumbi, is a native Australian plant, which has traditionally been used as an Aboriginal medicine. This study investigates the effect of different solvents and extractive fermentation on the content and natural products composition of Pittosporum angustifolium extracts, and compares their antioxidant activity, in vitro α-amylase inhibition, and anti-inflammatory properties. Anti-inflammatory activity of the extracts was determined by measuring the inhibition of nitric oxide (NO) production. Extracts were characterised with FTIR-ATR spectroscopy, and screened for antioxidant activities and α-amylase inhibitory activity via High-performance thin-layer chromatography (HPTLC)-Effect-directed analysis (EDA) with direct bioautography. HPTLC combined with chemical derivatization and bioassays was used for EDA screening. The results show that lactic acid fermentation of gumbi gumbi leaves boosts the antioxidant activity in extracts by increasing the total phenolic content, but does not affect (increase or decrease) α-amylase inhibitory activity or nitrogen scavenging/anti-inflammatory activity. Analysis of the ATR-FTIR spectra from the band at RF = 0.85 that inhibits α-amylase, suggests that fatty acid esters are responsible for the enzyme inhibition; both saturated fatty acid esters in unfermented extracts and unsaturated fatty acid esters in fermented extracts. The ATR-FTIR spectra of the polyphenolics in fermented extracts (RF = 0.15-0.20) suggests the presence of soluble lignin fragments (i.e. lignins depolymerized into monomers and oligomers during the fermentation process).


Asunto(s)
Antioxidantes/análisis , Extractos Vegetales/análisis , Rosales/química , Antioxidantes/química , Antioxidantes/farmacología , Bioensayo , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , Fermentación , Extractos Vegetales/química , Extractos Vegetales/farmacología , alfa-Amilasas/antagonistas & inhibidores
14.
J Pharm Biomed Anal ; 184: 113208, 2020 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-32114161

RESUMEN

Many native Australian plants have a long history of use as medicinal and culinary herbs and some are considered to be equivalents to the Mediterranean herbs. However, while therapeutic properties of Mediterranean herbs are well documented, there is limited information on the medicinal use of the Australian native herbs. Extracts of five native Australian plants were characterised with FTIR-ATR spectroscopy in the fingerprint region and screened for enzyme inhibitory and antioxidant activities via effect-directed analysis (EDA) based on bioautography. High-performance thin-layer chromatography (HPTLC) coupled with microchemical and biochemical derivatization assays was used for EDA screening. Detected compounds with biological activities were identified via FTIR-ATR spectroscopy. All herbs showed antioxidant activity with lemon myrtle being the most active. The α-amylase inhibition, observed in native thyme, sea parsley and native bush was associated with the presence of phenolic acids, chlorogenic acid and caffeic acid. The investigation of botanicals by a fast, hyphenated HPTLC method, has allowed an effect-directed high-throughput screening, fast characterization of complex mixtures and detection of biologically active phytochemicals (bioprofiling).


Asunto(s)
Bioensayo/métodos , Cromatografía Líquida de Alta Presión/métodos , Cromatografía en Capa Delgada/métodos , Extractos Vegetales/análisis , Extractos Vegetales/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Australia , Fitoquímicos/química
15.
J Chromatogr A ; 1620: 460970, 2020 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-32089291

RESUMEN

This study focuses on the health benefits of several fresh herbs that are commonly used in the Mediterranean diet. Antioxidant activity, phytosterol content and α-amylase inhibitory activity of fresh basil, lavender, oregano, rosemary, sage, and thyme are analyzed and compared. High-performance thin-layer chromatography (HPTLC) combined with effect directed analysis was used to detect and quantify biological active compounds on chromatograms. The highest antioxidant activity was measured in the extract from oregano leaf, while the highest terpenoid content was in basil leaf extract. All extracts except lavender leaf and lavender flower extracts showed α-amylase inhibition. The same compound at hRF = 68 in basil, oregano, rosemary, sage, and thyme extracts was responsible for α-amylase inhibition. Combined with effect-directed assays and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, hyphenated HPTLC allowed a fast characterization of the active compound. ATR spectral analysis of this band tentatively identified oleanolic acid (or its derivative) to be responsible for the α-amylase inhibition.


Asunto(s)
Lamiaceae/química , Extractos Vegetales/química , alfa-Amilasas/antagonistas & inhibidores , Antioxidantes/farmacología , Bioensayo , Cromatografía en Capa Delgada , Inhibidores Enzimáticos/farmacología , Fitosteroles/análisis , Extractos Vegetales/farmacología , Espectroscopía Infrarroja por Transformada de Fourier , Terpenos/análisis
16.
RSC Med Chem ; 11(3): 349-357, 2020 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-33479640

RESUMEN

Donor biological materials and excipients of animal origin are important components in the production of biomedical cell products (BMCPs). Their quality ensures the stability, safety, effectiveness and purity of the final product. This review discusses quality requirements for biological excipients intended for the production of biomedical cell products, in terms of the necessary information that should be included in the BMCP registration dossier during state registration and is subject to expert assessment during quality control. Considering that there is currently no production of biomedical cell products in the Russian Federation, the authors considered international approaches to ensuring the safety of donor material and excipients for the manufacturing of human cell- and tissue-based products (BMCP analogues).

17.
J Biomol Struct Dyn ; 38(8): 2402-2411, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-31204906

RESUMEN

Aromatherapy with essential oils (EOs) has been linked to improvement of cognitive function in patients with dementia. In order to act systemically, active EO components must be absorbed through the skin, enter the systemic circulation, and cross the blood brain barrier (BBB). Thus, the aim of this work was to develop quantitative structure activity relationships (QSARs), to predict skin and blood barrier penetrative abilities of 119 terpenoids from EOs used in aromatherapy. The first model was based on experimentally measured skin permeability for 162 molecules, and the second model on BBB permeability for 138 molecules. Each molecule was encoded with 63 calculated molecular descriptors and an artificial neural network was used to correlate molecular descriptors to permeabilities. Developed QSAR models confirm that EOs components penetrate through the skin and across the BBB. Some well-known descriptors, such as log P (lipophilicity), molecular size and shape, dominated the QSAR model for BBB permeability. Compounds with the highest predicted BBB penetration were hydrocarbon terpenes with the smallest molecular size and highest lipophilicity. Thus, molecular size is a limiting factor for penetration. Compounds with the highest skin permeability have slightly higher molecular size, high lipophilicity and low polarity. Our work shows that a major disadvantage of novel multitarget compounds developed for the treatment of Alzheimer's disease is the size of molecules, which cause problems in their delivery to the brain. Therefore, there is a need for smaller compounds, which possess more desirable physicochemical properties and pharmacokinetics, in addition to targeted biological effects.Communicated by Ramaswamy H. Sarma.


Asunto(s)
Aromaterapia , Demencia , Aceites Volátiles , Barrera Hematoencefálica , Humanos , Permeabilidad , Relación Estructura-Actividad Cuantitativa
18.
Dis Markers ; 2019: 6741518, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31885741

RESUMEN

Human saliva is increasingly being used and validated as a biofluid for diagnosing, monitoring systemic disease status, and predicting disease progression. The discovery of biomarkers in saliva biofluid offers unique opportunities to bypass the invasive procedure of blood sampling by using oral fluids to evaluate the health condition of a patient. Saliva biofluid is clinically relevant since its components can be found in plasma. As salivary lipids are among the most essential cellular components of human saliva, there is great potential for their use as biomarkers. Lipid composition in cells and tissues change in response to physiological changes and normal tissues have a different lipid composition than tissues affected by diseases. Lipid imbalance is closely associated with a number of human lifestyle-related diseases, such as atherosclerosis, diabetes, metabolic syndromes, systemic cancers, neurodegenerative diseases, and infectious diseases. Thus, identification of lipidomic biomarkers or key lipids in different diseases can be used to diagnose diseases and disease state and evaluate response to treatments. However, further research is needed to determine if saliva can be used as a surrogate to serum lipid profiles, given that highly sensitive methods with low limits of detection are needed to discover salivary biomarkers in order to develop reliable diagnostic and disease monitoring salivary tests. Lipidomic methods have greatly advanced in recent years with a constant advance in mass spectrometry (MS) and development of MS detectors with high accuracy and high resolution that are able to determine the elemental composition of many lipids.


Asunto(s)
Biomarcadores/química , Lipidómica/métodos , Saliva/química , Humanos , Estilo de Vida , Límite de Detección , Espectrometría de Masas , Estrés Fisiológico
19.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1130-1131: 121808, 2019 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-31669631

RESUMEN

A new sample extraction protocol was developed for pharmacokinetic studies of dabigatran with high-performance liquid chromatography separation - electrospray ionization time-of-flight mass spectrometry analysis. After protein precipitation with acetonitrile, free dabigatran and its metabolites are separated into water phase by water-dichloromethane liquid-liquid extraction to purify the sample from proteins and endogenous lipophilic compounds. Chromatographic separation was achieved on an Agilent Zorbax SB-CN column (150 × 4.6 mm, 5 µm)) using 0.1% aqueous solution of formic acid and acetonitrile (80:20) as the mobile phase. Agilent Zorbax SB-CN column was selected to improve sample resolution and to avoided early elution of dabigatran previously seen when using a C18 column. The extended calibration curve was constructed from 5 to 1000 ng/L while precision and accuracy were assessed at four levels across the linear dynamic ranges. Within-run precision was <5.6% and the between-run precision was <3.9%. The method accuracy ranged from 89.8% to 104.4%. The developed method was successfully applied to 30 patient samples to evaluate antithrombotic efficacy and anticoagulant activity of dabigatran following knee endoprosthesis surgery.


Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Dabigatrán/sangre , Dabigatrán/aislamiento & purificación , Espectrometría de Masas en Tándem/métodos , Dabigatrán/farmacocinética , Monitoreo de Drogas , Humanos , Límite de Detección , Modelos Lineales , Reproducibilidad de los Resultados
20.
Mar Drugs ; 17(3)2019 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-30832418

RESUMEN

Marine organisms produce an array of biologically active natural products, many of which have unique structures that have not been found in terrestrial organisms. Hence, marine algae provide a unique source of bioactive compounds. The present study investigated 19 marine algae and one seagrass collected from Torquay beach, Victoria, Australia. High-performance thin-layer chromatography (HPTLC) hyphenated with microchemical (DPPH•, p-anisaldehyde, and Fast Blue B) and biochemical (α-amylase and acetylcholine esterase (AChE) enzymatic) derivatizations was used to evaluate antioxidant activity, presence of phytosterols and phenolic lipids, α-amylase and AChE inhibitory activities of extract components. Significant α-amylase and AChE inhibitory activities were observed in samples 2, 6, 8 and 10. Antioxidant activities in the samples were found to be correlated to phytosterol content (R² = 0.78), but was not found to be related to either α-amylase or AChE inhibitory activities. α-Amylase inhibitory activities were correlated to AChE inhibition (R² = 0.77) and attributed to the phytosterol content, based on the similar peak position in the chromatograms with the ß-sitosterol chromatogram. Samples 1, 8, and especially sample 20, were found to contain phenolic lipids (alkyl resorcinol derivatives) with significant antioxidant activities. The results suggest that these marine species have a significant number of bioactive compounds that warrant further investigation.


Asunto(s)
Productos Biológicos/farmacología , Extractos Vegetales/farmacología , Algas Marinas/química , Acetilcolinesterasa/metabolismo , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Australia , Productos Biológicos/química , Productos Biológicos/aislamiento & purificación , Compuestos de Bifenilo/química , Compuestos de Bifenilo/aislamiento & purificación , Compuestos de Bifenilo/farmacología , Inhibidores de la Colinesterasa/química , Inhibidores de la Colinesterasa/aislamiento & purificación , Inhibidores de la Colinesterasa/farmacología , Cromatografía Líquida de Alta Presión/métodos , Cromatografía en Capa Delgada/métodos , Pruebas de Enzimas , Fitosteroles/química , Fitosteroles/aislamiento & purificación , Fitosteroles/farmacología , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , alfa-Amilasas/antagonistas & inhibidores , alfa-Amilasas/metabolismo
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