Chemical characterization, in vivo safety profile and effects of the extract from unconventional food plant Acca sellowiana (O. BERG) Burret on endothelial cells under glucose overload.

"Guava" (Acca sellowiana) is an unconventional edible plant from Brazil. It is used in traditional medicine as an anti-diabetic; however, pharmacological studies on this plant are scarce. This study aimed to evaluate the chemical and safety profile of an aqueous A. sellowiana peel extract (ASPE) and its effects on endothelial EA.hy926 cells under glucose overload and in vivo (Artemia salina). An ethanolic extract from A. sellowiana peels (ASPEetOH) was also produced and characterized. Results showed that ASPE did not present in vivo toxicity, and it was found to contain high phenolic content and redox capacity. ASPE (50 µg/mL; 24 h) prevented oxidative stress and mitochondrial dysfunction, besides positively modulating Sirtuins 1 and 3, and prevented the increase of COX-2 and NF-kß expression levels in EA.hy926 cells under glucose overload. Chromatographic fractionation, metabolite profiling, spectroscopic and bioinformatics analyses revealed the presence of phenolic acids, flavan-3-ols, flavonols, flavones, flavanones, and anthocyanidins, displaying a diversity of compounds in the crude and fractionated ASPEetOH. This study provided evidence on the safety profile, chemical composition, and pharmacological activities of A. sellowiana.

Evaluating Alterations in Breast Cancer Patients after Recovery Via A PET/CT-Assisted Metabolomics Approach.

OBJECTIVE: Breast cancer is a mortal disease that causes many deaths, especially in women. Improved therapies could contribute positively to survival rates. Metabolomics is an important tool for monitoring the alterations of several metabolites in clinical cases. This study aimed to develop a metabolomics model to observe (via mass spectroscopy) metabolic alterations in patients who suffered from breast cancer (BC), both before and after their recovery. MATERIALS AND METHODS: Grades 1 and 2 invasive ductal carcinoma patients were evaluated based on their positron emission tomography/computed tomography results. Fourteen patients who had fully recovered from BC were subjected to metabolomics analysis. Plasma samples were extracted and analyzed via quadrupole time-of-flight mass tandem spectroscopy. A chemometrics analysis was performed in order to determine the statistically significant metabolites. All the metabolites were annotated via the mummichog algorithm. RESULTS AND DISCUSSION: According to the data analysis, glucose, ornithine, phenyalanine, some vitamins, and metabolites in the fatty acid metabolism were statistically altered after recovery of each patient. CONCLUSION: Untargeted metabolomics studies can be used to understand the etiopathogenesis of breast cancer, finding new biomarkers and alterations of metabolic pathways. After the tumor burden was removed, homeostasis was restored and the concentration of several metabolites began to normalize. This study elucidated the effects of breast cancer at the molecular level.

Periodontal disease metabolomics signatures from different biofluids: a systematic review.

BACKGROUND: Periodontitis is resulted from a complex interaction between genetics and epigenetics, microbial factors, and the host response. Metabolomics analyses reflect both the steady-state physiological equilibrium of cells or organisms as well as their dynamic metabolic responses to environmental stimuli. AIM OF REVIEW: This systematic review of the literature aimed to assess which low molecular weight metabolites are more often found in biological fluids of individuals with periodontitis compared to individuals with gingivitis or periodontal health. KEY SCIENTIFIC CONCEPTS OF REVIEW: All the included studies employed untargeted analysis. One or more biological fluids were analyzed, including saliva (n = 14), gingival crevicular fluid (n = 6), mouthwash (n = 1), serum (n = 3) and plasma (n = 1). Fifty-six main metabolites related to periodontitis have been identified in at least two independent studies by NMR spectroscopy or MS-based metabolomics. Saliva was the main biological fluid sampled. It is noteworthy that 14 metabolites of the 56 detected were identified as main metabolites in all studies that sampled the saliva. The majority of metabolites found consistently among studies were amino acids, organic acids and derivates: acetate, alanine, butyrate, formate, GABA, lactate, propionate, phenylalanine and valine. They were either up- or down-regulated in the studies or this information was not mentioned. The main metabolic pathway was related to phenylalanine, tyrosine and tryptophan biosynthesis. Metabolites more frequently found in individuals with periodontitis were related to both the host and to microorganism responses. Future studies are needed, and they should follow some methodological standards to facilitate their comparison.

Hybrid Nanoparticles of Proanthocyanidins from Uncaria tomentosa Leaves: QTOF-ESI MS Characterization, Antioxidant Activity and Immune Cellular Response.

Previous studies in Uncaria tomentosa have shown promising results concerning the characterization of polyphenols with leaves yielding more diverse proanthocyanidins and higher bioactivities values. However, the polyphenols-microbiota interaction at the colonic level and their catabolites avoid the beneficial effects that can be exerted by this medicinal plant when consumed. In this regard, a new generation of hybrid nanoparticles has demonstrated improvements in natural compounds' activity by increasing their bioavailability. In this line, we report a detailed study of the characterization of a proanthocyanidin-enriched extract (PA-E) from U. tomentosa leaves from Costa Rica using UPLC-QTOF-ESI MS. Moreover, two types of hybrid nanoparticles, a polymeric-lipid (F-1) and a protein-lipid (F-2) loaded with PA-E were synthesized and their characterization was conducted by dynamic light scattering (DLS), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FT-IR), high-resolution transmission electron microscopy (HR-TEM), and encapsulation efficiency (%EE). In addition, in vitro release, antioxidant activity through 2,2-diphenyl-1-picrylhidrazyl (DPPH) as well as in vivo delayed-type hypersensitivity (DTH) reaction was evaluated. Results allowed the identification of 50 different compounds. The PA-E loaded nanoparticles F-1 and F-2 achieved encapsulation efficiency of ≥92%. The formulations exhibited porosity and spherical shapes with a size average of 26.1 ± 0.8 and 11.8 ± 3.3 nm for F-1 and F-2, respectively. PA-E increased its release rate from the nanoparticles compared to the free extract in water and antioxidant activity in an aqueous solution. In vivo, the delayed-type hypersensitive test shows the higher immune stimulation of the flavan-3-ols with higher molecular weight from U. tomentosa when administered as a nanoformulation, resulting in augmented antigen-specific responses. The present work constitutes to our knowledge, the first report on these bioactivities for proanthocyanidins from Uncaria tomentosa leaves when administrated by nanosystems, hence, enhancing the cellular response in mice, confirming their role in immune modulation.

Proteomic dataset of Listeria monocytogenes exposed to sublethal concentrations of free and nanoencapsulated nisin.

The cellular proteins of L. monocytogenes exposed to free and liposome-encapsulated nisin at sublethal concentration were hydrolyzed by trypsin and examined by tandem mass spectrometry (MS/MS) to obtain proteomic data. In the present study, we use the STRING v11.05 database analyze the interactions among the 78 upregulated proteins from L. monocytogenes obtained after treatment with sublethal concentrations of free and nanoliposome-encapsulated nisin. As result, from the upregulated proteins by free nisin was determined a network with 140 edges with two relevant nodes, containing ribosomal proteins and transmembrane transport proteins (SecD and ABC transport system). These two sets of proteins present biological connection as a group, with strong interactions and are related to detoxification and other Listeria response mechanisms. In addition, a high amount of membrane proteins was identified in the free nisin treatment. On the other hand, in the interaction analysis of upregulated proteins by liposome-loaded nisin, was found 156 edges with a single protein network, the same observed in free nisin, related to ribosomal proteins. Therefore, according with this analysis, the encapsulation of nisin into liposomes cause upregulation of ribosomal and decrease of L. monocytogenes response proteins as compared with free nisin.

LC-MS bioanalytical method for simultaneous determination of latanoprost and minoxidil in the skin.

The association of minoxidil sulphate and latanoprost is currently emerging as a promising strategy for the treatment of androgenic alopecia, which is the most common type of scalp hair loss. In order to support the development of new pharmaceutical products containing such drugs combination, this study proposes a simple and efficient LC-MS bioanalytical method to simultaneously quantify minoxidil sulphate and latanoprost in different skin layers. Compounds separation was performed by liquid chromatography using a C18 column with gradient elution of a mobile phase composed of 0.1 % formic acid in acetonitrile and water at a flow rate of 0.5 mL min-1. Determinations were executed using mass spectrometry equipped with an ESI interface operating in a positive ionization mode. Quantification was performed using selective ion mode monitoring of m/z 210.1 for minoxidil sulphate and 433.3 for latanoprost. The matrix effect was very pronounced in samples containing some skin layers or electrolyte solution. Accordingly, a calibration curve for each contaminant group was built, leading to correlation coefficient values higher than 0.99. Additionally, lower limits of detection and quantification were obtained, and precision (repeatability and intermediate precision) achieved results with a coefficient of variation less than 15 %. Drug recovery from skin samples was higher than 70 %, fulfilling the recommendations. Also, the bioanalytical method was successfully tested in in vitro skin penetration studies proving its effectiveness in the development of topical formulations containing both drugs.

A critical review of HPLC-based analytical methods for quantification of Linezolid.

Linezolid is a synthetic antimicrobial agent belonging to the oxazolidinone class. Since its approval in the year 2000 until now, linezolid remains the main representative drug for the oxazolidinone class of drugs, which is used in therapy due to its unique mode of action, which involves inhibition of protein synthesis. As linezolid holds great importance in antimicrobial therapy, it is necessary to compile the various analytical methods that have been reported in the literature for its analysis. Analytical techniques used for pharmaceutical analyses and therapeutic drug monitoring play an important role in comprehending the aspects regarding bioavailability, bioequivalence, and therapeutic monitoring during patient follow-ups. Even though linezolid has had the approval for clinical use for more than 18 years now, most of the analytical methods for its determination reported in the scientific literature are the ones which utilize HPLC. Therefore, the present review provides a summary of the HPLC-based methods used in the determination and quantification of linezolid in different matrices since the time of its discovery.

Analysis of molecular changes induced by mineral trioxide aggregate on spla2

Abstract The aim of this study was to analyze the effects of MTA on the structure and enzymatic activity of sPLA2 in order to provide subsidies for improvement in the formulation of the product. MTA powder was incubated for 60 min in the presence of sPLA2 and was analyzed by chromatography, electrospray mass (ESI-MS) and small-angle X-ray scattering (SAXS). It was find that the elution profile, retention time, and fragmentation of sPLA2 were altered after treatment with MTA. Calcium was the MTA component that most amplified the inflammatory signal. Significant interactions were found between MTA and sPLA2, which could aid in our understanding of the mechanisms of action of MTA during the inflammatory process and it may facilitate the structural modification of MTA, thereby improving its biological safety and consequently the rate of the treatment success.

Resumo O objetivo deste estudo foi analisar os efeitos do MTA na estrutura e atividade enzimática da sPLA2 a fim de fornecer subsídios para melhoria na formulação do produto. O MTA em pó foi incubado por 60 min na presença de sPLA2 e analisado por cromatografia, espectroscopia de massa por eletropulverização (ESI-MS) e espalhamento de raios-X de baixo ângulo (SAXS). Encontrou-se que o perfil de eluição, o tempo de retenção e a fragmentação da sPLA2 foram alterados após o tratamento com MTA. O cálcio foi o componente do MTA que mais ampliou o sinal inflamatório. Encontraram-se interações significativas entre o MTA e o sPLA2, o que poderia auxiliar na compreensão dos mecanismos de ação do MTA durante o processo inflamatório e facilitar a modificação estrutural do MTA, melhorando sua segurança biológica e consequentemente a taxa de sucesso do tratamento.

Discrimination between Asian populations of the parasitoid wasp Ganaspis cf. brasiliensis using a simple MALDI-TOF MS-based method for use with insects.

The fruit fly Drosophila suzukii has recently become an invasive pest insect of significant economic impact in Europe and the USA. In contrast to other Drosophila species, D. suzukii is able to infest intact fruit by means of a saw-like ovipositor, which allows females to deposit eggs beneath the skin of the fruit. Classical biological control using the parasitoid wasp Ganaspis cf. brasiliensis is currently being researched as an environmentally sustainable option for the control of D. suzukii. In particular, the host specificity of this parasitoid has been assessed for populations from different regions in China and Japan. In order to study the relationship between the differences in specificity and molecular variations, we have adapted a matrix-assisted laser-desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based method, originally developed for use with plant material, to discriminate between example populations of G. cf. brasiliensis. We have employed a combination of principal component analysis and blind-tested comparison between reference sample MALDI-TOF MS spectra and test sample spectra to discriminate, on the basis of the acid-soluble insect protein spectra generated, between four populations of G. cf. brasiliensis (originally collected from Tokyo and Hasuike in Japan and Dali and Ximing in China). MALDI-TOF MS analysis is able to discriminate with 100% accuracy between populations G. cf. brasiliensis. The Chinese populations were observed to be similar, but the Tokyo population is slightly different and the Hasuike population is significantly different from the other populations. The Tokyo population appears more closely related to the Chinese populations than the Hasuike population, even though both originate from Japan.

Determination of fatty acid methyl esters in cosmetic castor oils by flow injection - electrospray ionization - high resolution mass spectrometry.

OBJECTIVE: The goal of this work was to set up a high throughput procedure for the determination of fatty acid methyl esters (FAMEs) in cosmetic castor oils using flow injection - electrospray ionization - high resolution mass spectrometry, and to demonstrate the need of such analysis for the quality control purposes. METHODS: The sample aliquot was mixed with isooctane:chloroform (1:1) and submitted to transesterification; the obtained FAMEs were appropriately diluted using water:isopropanol:acetonitrile (20:50:30) with addition of sodium formate which served as an internal standard, lock mass calibrant and promoted the formation of sodium adducts during electrospray ionization (ESI). The principle of flow injection analysis (FIA) was applied for sample introduction to an ESI - quadrupole- time of flight mass spectrometer (ESI-QTOFMS). The carrier solution was composed of water:isopropanol:acetonitrile (20:50:30). From the acquired MS data, flowgrams of the extracted [M+Na]+ ions were obtained using the following m/z values for individual FAMEs: 293.2451 (C16:0); 315.2295 (C18:3); 317.2451 (C18:2); 319.2608 (C18:1); 321.2764 (C18:0); 335.2557 (C18:1,OH); 349.3077 (C20:0); 377.3390 (C22:0) and m/z 226.9515 for IS. Baseline-subtracted and filtered signals were integrated and the list of peaks intensities was exported to Excel, where calibration functions were obtained and quantification carried out. Gas chromatography with a flame ionization detector (GC-FID) was used as an alternative analytical tool. RESULTS: The calibration detection limits for FAMEs of unsaturated fatty acids were in the range 3.61 - 8.62 µg L-1 and for saturated compounds in the range 8.51 - 82.4 µg L-1 . The results obtained for commercial were in good agreement with GC-FID data; among nine cosmetic oils analyzed, three contained low concentrations of ricinoleic acid (C18:1, OH), indicating adulteration of castor bean oil with other vegetable oils. CONCLUSION: Application of FIA for the sample introduction to ESI-QTOFMS enabled for reliable determination of FAMEs in cosmetic oils with sampling frequency of thirty per hour as compared to two samples per hour achievable using GC-FID. The proposed procedure is especially well suited for FAMEs of unsaturated fatty acids that are primary components of castor triacylglycerides, and contribute to desirable properties of any cosmetic oil. This article is protected by copyright. All rights reserved.

Estudio metabolómico de la enfermedad renal crónica: del modelo experimental al humano / Metabolomics in chronic kidney disease: from experimental model to human disease

Con una prevalencia global reportada de entre 11-13%, la enfermedad renal crónica (ERC) ha sido reconocida como un gran desafío para los sistemas de salud por sus implicaciones económicas y sociales. Al tratarse de una enfermedad crónica e irreversible, el tratamiento está dirigido a disminuir su progresión. La cuantificación de creatinina sérica es el método de elección para su diagnóstico y clasificación; sin embargo, es conocido que esta prueba tiene una sensibilidad clínica limitada, lo que ha conducido a la búsqueda de nuevos marcadores que permitan un diagnóstico y monitoreo oportuno. Desde esta perspectiva, el empleo de la metabolómica y de modelos animales ha permitido la identificación y estudio de nuevos metabolitos, candidatos a ser utilizados como futuros biomarcadores en la práctica clínica. La presente revisión tuvo como objetivo hacer una análisis de los perfiles metabolómicos reportados para la ERC, tanto en modelos experimentales como en estudios realizados en seres humanos. De acuerdo a los datos obtenidos, los metabolitos implicados en las rutas metabólicas de aminas cuaternarias y aminoácidos como el TMNO, el indoxilsulfato y derivados de la dimetilarginina representan una alternativa prometedora para la identificación, clasificación y pronóstico de la ERC.

Chronic kidney disease (CKD) high global prevalence, estimated between 11 to 13%, has been recognized as a mayor health challenge for healthcare systems due to its relevant economic and social implications. Main medical intervention strategies are directed to delay the progression of CKD and prevent outcomes. Serum creatinine concentration has been used to classify CKD and define its progression stage; however, it is well known the low sensitivity shown by this test. This fact has conducted to the search for new markers in order to improve the disease diagnosis, monitoring and treatment. In this context, metabolomics science and animal models have allowed identification of new metabolites that can be used as future biomarkers into clinical practice. This review aims to summarize the metabolomics profiles reported in different experimental models and clinical research on CKD. According with the data obtained, metabolites related with quaternary amines and aminoacid metabolic pathways like TMNO, indoxyl sulfate and dimethylarginine, suggest a promising alternative for identification, classification and prognosis of CKD.

Isoniazid: A Review of Characteristics, Properties and Analytical Methods.

Isoniazid is a synthetic antimicrobial and one of the most important first-line drugs used in the treatment of tuberculosis. Since it was introduced in the therapy in 1952, the drug remains at the front line of the antituberculosis treatment mainly due to its potency and high selectivity against Mycobacterium tuberculosis. Pharmaceutical analysis and therapeutic drug monitoring of isoniazid in both, pharmaceuticals and biological samples, plays an important role to comprehend aspects regarding to bioavailability, bioequivalence and therapeutic monitoring during patients following-up. In the last case, validated and simple methods are extremely useful for Public Healthy in order to guarantee the drug efficacy, safety and reduce the tuberculosis resistance. Among the available analytical tools, HPLC-based methods coupled to ultraviolet or mass spectroscopy are the most widely used techniques to quantify isoniazid. Therefore, this review highlights the main analytical methods reported in the literature for determination of isoniazid focusing in HPLC-based methods.

Physicochemical and antioxidant characterization of Justicia spicigera.

Extracts with water:ethanol (100:0, 70:30, 50:50, 30:70, 0:100) solutions from fresh (F), just dried (JD), dried and stored for one year (DS) Justicia spicigera leaves were obtained using the stirring and ultrasound techniques. Extracts were analyzed in physicochemical and antioxidant characteristics. Identification of chemical compounds by gas chromatography-mass spectroscopy (GC-MS) was also performed. 2.14±0.91, 5.67±1.70, and 8.52±4.97g Gallic acid equivalents/100g dry weight (d.w.) of phenolic compounds were found, in average, for F, JD, and DS J. spicigera, respectively. 2.22±1.31, 2.58±2.11, and 8.48±3.78g Trolox equivalents/100g d.w. were detected with the ABTS method and 0.49±0.33, 1.23±0.87, and 0.88±0.94g with the DPPH method for F, JD and DS J. spicigera, respectively. Eucalyptol, phytol, and azulene were identified as the main compounds. J. spicigera showed colors (green-iridescent, green-yellow, or pink of different intensities) and antioxidant characteristics depending on the solvent concentration. Extracts could be used in the food and pharmaceutical industries.

Hydroalcoholic extract of red propolis promotes functional recovery and axon repair after sciatic nerve injury in rats.

CONTEXT: Peripheral axon injury and degeneration are often mediated by oxidative stress and inflammation. The hydroalcoholic extract of the red propolis (HERP) has attracted great attention because of its antioxidant and anti-inflammatory activities. OBJECTIVE: The objective of this work is to study the effect of HERP on nerve repair and functional recovery after sciatic nerve injury (SNI) in rats. MATERIALS AND METHODS: The chemical markers in HERP were identified using high-resolution mass spectroscopy. After axonotmesis of sciatic nerve, ibuprofen (IBP) and HERP treatments were orally administered for 28 d. Behavioural tests were performed weekly after SNI. The myelinated axon number was counted using morphometric analysis. RESULTS: The compounds found in HERP were pinocembrin, formononetin, vestitol, and biochanin A. The animals that underwent SNI showed a significant decrease in motor function based on the Basso, Beattie and Bresnahan scale and sciatic functional index compared with sham animals until 7 d after the surgery (p < 0.05). After 14 and 21 d, the SNI groups treated with either HERP or IBP showed significant improvement (p < 0.01), and the SNI group treated with HERP 10 mg/kg showed accelerated motor recovery compared with the other groups (p < 0.01). SNI caused also a reduction in the myelinated axon counts, and treatment with HERP 10 mg/kg induced a significant increase in the number of myelinated fibres compared with all other groups. CONCLUSION: HERP promoted regenerative responses and accelerated functional recovery after sciatic nerve crush. Thus, it can be considered to be a new strategy or complementary therapy for treating nerve injuries.

Ultra-fast determination of caffeine, dipyrone, and acetylsalicylic acid by capillary electrophoresis with capacitively coupled contactless conductivity detection and identification of degradation products.

Capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C(4)D) was used for fast, simultaneous determination of dipyrone (DIP), caffeine (CAF), and acetylsalicylic acid (ASA). In the same run and in less than 1min, the degradation products from DIP and ASA were also detected. In addition, the usage of the CE-C(4)D system allowed, for the first time, the detection of methylamine as a degradation product of DIP. Capillary electrophoresis with electrospray mass spectrometry experiments were carried out in order to confirm the formation of methylamine. The limits of detection by CE-C(4)D were 5, 5, and 6µmolL(-1) for CAF, DIP, and ASA, respectively. The proposed method was applied to the analysis of these compounds in pharmaceutical formulations with similar results to those achieved by HPLC (p<0.05).

Recent advances in maize nuclear proteomic studies reveal histone modifications.

The nucleus of eukaryotic organisms is highly dynamic and complex, containing different types of macromolecules including DNA, RNA, and a wide range of proteins. Novel proteomic applications have led to a better overall determination of nucleus protein content. Although nuclear plant proteomics is only at the initial phase, several studies have been reported and are summarized in this review using different plants species, such as Arabidopsis thaliana, rice, cowpea, onion, garden cress, and barrel clover. These include the description of the total nuclear or phospho-proteome (i.e., Arabidopsis, cowpea, onion), or the analysis of the differential nuclear proteome under different growth environments (i.e., Arabidopsis, rice, cowpea, onion, garden cress, and barrel clover). However, only few reports exist on the analysis of the maize nuclear proteome or its changes under various conditions. This review will present recent data on the study of the nuclear maize proteome, including the analysis of changes in posttranslational modifications in histone proteins.

Structure elucidation of a flavonoid glycoside from the roots of Clerodendrum serratum (L.) Moon, Lamiaceae

Apigenin-7-glucoside, C21H20O10 (7-(β-D-glucopyranosyloxy)-5-hydroxy-2-(4-hydroxyphenyl)-4H-1-benzopyran-4-one), was first time isolated from the roots of Clerodendrum serratum (L.) Moon, Lamiaceae. Structure elucidation of the compound was carried out by ¹H NMR and FAB-MS studies.

Apigenin-7-glucosídeo, C21H20O10 (7-(β-D-glucopiranosiloxi)-5-hidroxi-2-(4-hidroxifenil)-4H-1-benzopiran-4-ona), foi isolado pela primeira vez das raízes de Clerodendrum serratum (L.) Moon, Lamiaceae. A elucidação estrutural da susbtância foi feita através de estudos de ¹H NMR e FAB-MS.

Mass spectroscopy investigation of the effect of gamma irradiation on the mean value of the number of ethoxy groups in the triton x-100 / Estudio del efecto de la radiación gamma en elvalor medio del número de grupos etoxiLados del surfactante Tritón X-100

El efecto de la radiación gamma proveniente de una fuente de 60Co en la estructura del surfactante no iónico Tritón X-100 fue investigado. Tres regiones principales pueden ser distinguidas en el comportamiento del valor medio del número de grupos etóxidos al aumentar la dosis. Sin embargo, el resultado global encontrado fue una pequeña variación en este valor medio al cambiar la dosis entre 0 y 70 KGy.

The effect of gamma radiation from a 60Co source on the structure of a nonionic surfactant, namely TRITON X-100, was investigated. Three main regions can be distinguished in the behavior of the mean value of ethoxy groups with an increase in the absorbed dose. However just a slightly decrease on this mean value was obtained when the dose range from 0 to 70 kGy.