RESUMO
E-liquids have become increasingly popular in society in recent years. A wide variety of flavors and nicotine strengths make it possible for every user to get a product according to their wishes. Many of these e-liquids are marketed with countless different flavors, which are often characterized by a strong and sweet smell. Sweeteners, such as sucralose, are therefore commonly added as sugar substitutes. However, recent studies have shown the potential formation of highly toxic chlorinated compounds. This can be explained by the high temperatures (above 120 °C) within the heating coils and the used basic composition of these liquids. Nevertheless, the legal situation is composed of proposals without clear restrictions, only recommendations for tobacco products. For this reason, a high level of interest lies within the establishment of fast, reliable and cost-effective methods for the detection of sucralose in e-liquids. In this study, a number of 100 commercially available e-liquids was screened for sucralose in order to identify the suitability of ambient mass spectrometry and near-infrared spectroscopy for this application. A highly sensitive high-performance liquid chromatography coupled to a tandem mass spectrometer method was used as reference method. Furthermore, the advantages and limitations of the two mentioned methods are highlighted in order to provide a reliable quantification of sucralose. The results clearly revile the necessity for product quality due to the absence of declaration on many of the used products. Further on, it could be shown, that both methods are suitable for the quantification of sucralose in e-liquids, with beneficial economic and ecological aspects, over classical analytical tools including high-performance liquid chromatography. Clear correlations between the reference and novel developed methods are displayed. In summary, these methods enable an important contribution to ensure consumer protection and elimination of confuse package labelling.
Assuntos
Sistemas Eletrônicos de Liberação de Nicotina , Espectroscopia de Luz Próxima ao Infravermelho , Sacarose/análise , Edulcorantes/análise , Espectrometria de MassasRESUMO
The interaction of water with polymers is an intensively studied topic. Vibrational spectroscopy techniques, mid-infrared (MIR) and Raman, were often used to investigate the properties of water-polymer systems. On the other hand, relatively little attention has been given to the potential of using near-infrared (NIR) spectroscopy (12,500-4000 cm-1; 800-2500 nm) for exploring this problem. NIR spectroscopy delivers exclusive opportunities for the investigation of molecular structure and interactions. This technique derives information from overtones and combination bands, which provide unique insights into molecular interactions. It is also very well suited for the investigation of aqueous systems, as both the bands of water and the polymer can be reliably acquired in a range of concentrations in a more straightforward manner than it is possible with MIR spectroscopy. In this study, we applied NIR spectroscopy to investigate interactions of water with polymers of varying hydrophobicity: polytetrafluoroethylene (PTFE), polypropylene (PP), polystyrene (PS), polyvinylchloride (PVC), polyoxymethylene (POM), polyamide 6 (PA), lignin (Lig), chitin (Chi) and cellulose (Cell). Polymer-water mixtures in the concentration range of water between 1-10%(w/w) were investigated. Spectra analysis and interpretation were performed with the use of difference spectroscopy, Principal Component Analysis (PCA), Median Linkage Clustering (MLC), Partial Least Squares Regression (PLSR), Multivariate Curve Resolution Alternating Least Squares (MCR-ALS) and Two-Dimensional Correlation Spectroscopy (2D-COS). Additionally, from the obtained data, aquagrams were constructed and interpreted with aid of the conclusions drawn from the conventional approaches. We deepened insights into the problem of water bands obscuring compound-specific signals in the NIR spectrum, which is often a limiting factor in analytical applications. The study unveiled clearly visible trends in NIR spectra associated with the chemical nature of the polymer and its increasing hydrophilicity. We demonstrated that changes in the NIR spectrum of water are manifested even in the case of interaction with highly hydrophobic polymers (e.g., PTFE). Furthermore, the unveiled spectral patterns of water in the presence of different polymers were found to be dissimilar between the two major water bands in NIR spectrum (νs + νas and νas + δ).
Assuntos
Lignina , Água , Celulose , Quitina , Polímeros , Polipropilenos , Poliestirenos , Politetrafluoretileno , Cloreto de Polivinila , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Água/químicaRESUMO
Recently polyphenols attracted great interest in the field of food and nutrition as well as in the pharmaceutical and cosmetics industries due to their health benefits through antioxidative behavior in the human body. However, because of the high number of compounds characterized as phenols and their structural diversity, quantification of polyphenols turns out to be a highly complex task. Although, a wide variety of analytical methods are used for the determination of total polyphenolic content, they are all found to be lacking in a variety of different tasks, such as their limits of detection and quantification, repeatability, accuracy and specificity. For this reason, a novel approach combining the advantages of solid phase purification, near infrared analysis and multivariate data analysis was investigated for the prediction of total polyphenolic content, suitable for a wide range of sample matrices. Dispersive solid phase extraction was performed and optimized using polyvinylpyrrolidone as sorbent, known to selectively bind polyphenols. Near-infrared detection of adsorbed polyphenols was carried out subsequently. Furthermore, the method was in-house validated, examining selectivity, repeatability and accuracy, working range, as well as multivariate limit of detection and limit of quantification, comparing it with two routinely used methods-namely, Folin-Ciocalteu photometric assay and Löwenthal titration. The novel established method was applied for the prediction of total polyphenolic content in tea and wine samples.
Assuntos
Polifenóis/isolamento & purificação , Povidona/química , Extração em Fase Sólida , Antioxidantes/química , Humanos , Polifenóis/químicaRESUMO
Mid-infrared (MIR) microscopic imaging of indolent and aggressive lymphomas was performed including formalin-fixed and paraffin-embedded samples of six follicular lymphomas and 12 diffuse large B-cell-lymphomas as well as reactive lymph nodes to investigate benefits and challenges for lymphoma diagnosis. MIR images were compared to defined pathological characteristics such as indolent versus aggressive versus reactive, germinal centre versus activated cell-of-origin (COO) subtypes, or a low versus a high proliferative index and level of PD-L1 expression. We demonstrated that MIR microscopic imaging can differentiate between reactive lymph nodes, indolent and aggressive lymphoma samples. Also, it has potential to be used in the subtyping of lymphomas, as shown with the differentiation between COO subtypes, the level of proliferation and PD-L1 expression. MIR microscopic imaging is a promising tool for diagnosis and subtyping of lymphoma and further evaluation is needed to fully explore the advantages and disadvantages of this method for pathological diagnosis.
Assuntos
Linfoma Difuso de Grandes Células B , Humanos , Linfonodos/diagnóstico por imagem , Linfoma Difuso de Grandes Células B/diagnóstico por imagemRESUMO
The goal of this project is to identify any in-depth benefits and drawbacks in the diagnosis of amalgam tattoos and other pigmented intraoral lesions using hyperspectral imagery collected from amalgam tattoos, benign, and malignant melanocytic neoplasms. Software solutions capable of classifying pigmented lesions of the skin already exist, but conventional red, green and blue images may be reaching an upper limit in their performance. Emerging technologies, such as hyperspectral imaging (HSI) utilize more than a hundred, continuous data channels, while also collecting data in the infrared. A total of 18 paraffin-embedded human tissue specimens of dark pigmented intraoral lesions (including the lip) were analyzed using visible and near-infrared (VIS-NIR) hyperspectral imagery obtained from HE-stained histopathological slides. Transmittance data were collected between 450 and 900 nm using a snapshot camera mounted to a microscope with a halogen light source. VIS-NIR spectra collected from different specimens, such as melanocytic cells and other tissues (eg, epithelium), produced distinct and diagnostic spectra that were used to identify these materials in several regions of interest, making it possible to distinguish between intraoral amalgam tattoos (intramucosal metallic foreign bodies) and melanocytic lesions of the intraoral mucosa and the lip (each with P < .01 using the independent t test). HSI is presented as a diagnostic tool for the rapidly growing field of digital pathology. In this preliminary study, amalgam tattoos were reliably differentiated from melanocytic lesions of the oral cavity and the lip.
Assuntos
Transtornos da Pigmentação , Tatuagem , Humanos , Imageamento Hiperespectral , Melanócitos , MicroscopiaRESUMO
While a shift in energy metabolism is essential to cancers, the knowledge about the involvement of the mitochondrial genome in tumorigenesis and progression in oral squamous cell carcinoma (OSCC) is still very limited. In this study, we evaluated 37 OSCC tumors and the corresponding benign mucosa tissue pairs by deep sequencing of the complete mitochondrial DNA (mtDNA). After extensive quality control, we identified 287 variants, 137 in tumor and 150 in benign samples exceeding the 1% threshold. Variant heteroplasmy levels were significantly increased in cancer compared to benign tissues (p = 0.0002). Furthermore, pairwise high heteroplasmy frequency difference variants (∆HF% > 20) with potential functional impact were increased in the cancer tissues (p = 0.024). Fourteen mutations were identified in the protein-coding region, out of which thirteen were detected in cancer and only one in benign tissue. After eight years of follow-up, the risk of mortality was higher for patients who harbored at least one ∆HF% > 20 variant in mtDNA protein-coding regions relative to those with no mutations (HR = 4.6, (95%CI = 1.3-17); p = 0.019 in primary tumor carriers). Haplogroup affiliation showed an impact on survival time, which however needs confirmation in a larger study. In conclusion, we observed a significantly higher accumulation of somatic mutations in the cancer tissues associated with a worse prognosis.
RESUMO
The aim of this study was to improve intestinal mucus permeation of a peptide antibiotic via incorporation into papain-palmitate-modified self-emulsifying drug delivery systems (SEDDS) as nanocarrier. Vancomycin as a peptide antibiotic was lipidized by hydrophobic ion pair formation using sodium bis-2-ethylhexyl-sulphosuccinate before incorporation in SEDDS comprising Capmul MCM, propylenglycol, and Kolliphor EL (2:1:2). As mucolytic agent, 0.5% papain-palmitate was introduced in SEDDS formulation containing the vancomycin-sodium bis-2-ethylhexyl-sulphosuccinate ion pair. The formulation was evaluated regarding droplet size, zeta potential, and cytotoxicity using Caco-2 cells previous to intestinal mucus permeation studies using Transwell diffusion and rotating tube method. The hydrophobic ion pair product yielded from surfactant to drug ratio of 3:1 provided a 25-fold increase in lipophilicity, drug payload in SEDDS of 5%, and log DSEDDS/release medium of 2.2. The formulation exhibited a droplet size and zeta potential of 221.5 ± 14.8 nm and -4.2 ± 0.8 mV, respectively. Cytotoxicity study showed that SEDDS formulations were not toxic. Introducing 0.5% papain-palmitate increased the mucus permeability of SEDDS 2.8-fold and 3.3-fold in Transwell diffusion and rotating tube studies, respectively. According to these results, papain decorated SEDDS might be a potential strategy to improve the mucus permeating properties of peptide antibiotics.
Assuntos
Portadores de Fármacos/química , Mucosa Intestinal/metabolismo , Nanopartículas/química , Palmitatos/química , Papaína/química , Permeabilidade/efeitos dos fármacos , Vancomicina/química , Células CACO-2 , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos/métodos , Emulsificantes/química , Emulsões/química , Humanos , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Vancomicina/metabolismoRESUMO
BACKGROUND: Dark pigmented lesions of the oral mucosa can represent a major diagnostic challenge. A biopsy is usually required to determine the nature of such intraoral discolorations. This study investigates the potential use of infrared spectroscopy for differential diagnosis of amalgam tattoos versus benign or malignant melanocytic neoplasms. MATERIALS AND METHODS: For this retrospective study, formalin-fixed paraffin-embedded tissue (FFPE) specimens of dark pigmented lesions concerning the oral mucosa or the lip were investigated using mid infrared spectroscopy. The samples were chosen from patients who had undergone a mucosal biopsy at the University Hospital Innsbruck (Austria) between the years 2000 and 2017. Principal component analysis was used for data exploration. Evaluation was based on the superimposition of the recorded spectra and the corresponding histologic slides. RESULTS: In total, 22 FFPE specimens were analyzed. Clear differences were found between amalgam and non-amalgam samples. A general weakening of the penetrating infrared radiation allowed for unspecific discrimination between these two classes. An overall accuracy in predicting the correct class of 95.24% was achieved. CONCLUSION: Infrared spectroscopy appears to be a suitable technique to differentiate between amalgam tattoos and melanocytic lesions in FFPE samples. It could potentially be applied in vivo, too, serving as a non-invasive diagnostic tool for intraoral dark pigmented lesions.
Assuntos
Melanoma/diagnóstico , Mucosa Bucal/química , Espectrofotometria Infravermelho , Tatuagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Complexos de Coordenação/química , Diagnóstico Diferencial , Análise Discriminante , Feminino , Humanos , Masculino , Melanoma/patologia , Pessoa de Meia-Idade , Mucosa Bucal/patologia , Pigmentação , Análise de Componente Principal , Estudos RetrospectivosRESUMO
The performance of three portable NIR spectrometers was compared by analysing the total antioxidant capacity (TAC) of different species of gluten-free grains. TAC is often used to evaluate the quality of foods and was determined using Folin-Ciocalteu measurements and used as reference data for establishing PLS-R models with NIR data. NIRS enables fast and non-invasive measurements. The microPhazir RX and the MicroNIR 2200 are broadly used in chemical and pharmaceutical industries, whereas SCiO is a pocket-sized, consumer-oriented spectrometer. The devices work in different regions of the NIR spectrum and their performances was compared using statistical parameters. 77 samples were measured and analysed using the software The Unscrambler X, as well as SCiO-Lab. All models established were cross- and test set validated. The multivariate data processing using The Unscrambler X yielded similar results as SCiO-Lab. The best model was established for non-milled samples measured with the MicroNIR 2200 and analysed using The Unscrambler X.
Assuntos
Antioxidantes/análise , Grão Comestível/química , Espectroscopia de Luz Próxima ao Infravermelho/instrumentação , Antioxidantes/química , Glutens/análise , Análise Multivariada , Padrões de Referência , Espectroscopia de Luz Próxima ao Infravermelho/normasRESUMO
This study aimed to extract the paraffin component from paraffin-embedded oral cancer tissue spectra using three multivariate analysis (MVA) methods; Independent Component Analysis (ICA), Partial Least Squares (PLS) and Independent Component - Partial Least Square (IC-PLS). The estimated paraffin components were used for removing the contribution of paraffin from the tissue spectra. These three methods were compared in terms of the efficiency of paraffin removal and the ability to retain the tissue information. It was found that ICA, PLS and IC-PLS could remove the paraffin component from the spectra at almost the same level while Principal Component Analysis (PCA) was incapable. In terms of retaining cancer tissue spectral integrity, effects of PLS and IC-PLS on the non-paraffin region were significantly less than that of ICA where cancer tissue spectral areas were deteriorated. The paraffin-removed spectra were used for constructing Raman images of oral cancer tissue and compared with Hematoxylin and Eosin (H&E) stained tissues for verification. This study has demonstrated the capability of Raman spectroscopy together with multivariate analysis methods as a diagnostic tool for the paraffin-embedded tissue section.
Assuntos
Técnicas Citológicas , Neoplasias/diagnóstico , Análise Espectral Raman , Adulto , Idoso , Humanos , Pessoa de Meia-Idade , Análise Multivariada , Neoplasias/patologia , Inclusão em Parafina , Análise de Componente Principal , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise Espectral Raman/métodosRESUMO
During the last years, non-invasive or minimally invasive diagnostic tools in cancer diagnostics have become more important. Many fluorescence spectroscopic methodologies have been established for nearly all different kinds of cancer. The reason therefore is its high sensitivity, low amount of sample required, short testing time, and the suitability for in situ testing. The potential influence factors for cancer diagnostics and the subsequent suitability of the method to different applications are well described. Near-Infrared spectroscopy (NIRS) is based on differences of endogenous chromophores between cancer and normal tissues using either oxyhaemoglobin or deoxy-haemoglobin, lipid or water bands, or a combination of two or more of these diagnostic markers. These marker bands are known to provide the fundamental for the diagnosis of several cancers and the spectroscopic setup can be applied for the analysis of cells, urine and tissue. For the preparation of this review the literature published during the last fifteen years has been taken into consideration. It will provide an overview on the importance of the fluorescence and NIRS tools in cancer analysis giving hints about how these techniques can play a crucial role in cancer diagnosis, treatment decisions and therapy. The two techniques, fluorescence and near-infrared spectroscopy (NIRS) are faced to each other and individual advantages and/or drawbacks are discussed. Finally, it will be taken into consideration; how the synergistic combination of different approaches can give additional information related to development and progression stages of cancer.
Assuntos
Neoplasias/diagnóstico , Células-Tronco Neoplásicas/química , Saliva/química , Soro/química , Urina/química , Biomarcadores Tumorais/química , Análise por Conglomerados , Humanos , Neoplasias/química , Neoplasias/metabolismo , Células-Tronco Neoplásicas/citologia , Células-Tronco Neoplásicas/metabolismo , Espectrometria de Fluorescência , Espectroscopia de Luz Próxima ao InfravermelhoRESUMO
OBJECTIVES: To develop a self nano-emulsifying delivery system (SNEDS) for model peptide lanreotide providing a protective effect towards thiol-disulfide exchange reactions. METHODS: Ion-paired complexes of lanreotide with surfactants were prepared. In the following, Log P (octanol/water) of these complexes was determined. Lanreotide-loaded SNEDS (Lan/Deo-SN2 and Lan/Deo-SN3) were characterized for payload, droplet size and zeta potential. Lan/Deo-SN2 and Lan/Deo-SN3 were incubated with reduced glutathione (GSH) and thiol-enriched casein peptones for the assessment of thiol-disulfide exchange reactions. Ultra-centrifugation was used for separation of lanreotide released from SNEDS. RESULTS: A maximum payload of 6.4% was achieved for Lan/Deo-SN2. Mean droplet size of Lan/Deo-SN2 and Lan/Deo-SN3 was 45 ± 0.20 nm and 37 ± 0.02 nm, respectively. Both formulations showed significant protection towards thiol-disulfide exchange reactions. After 3 h of incubation with GSH, 48% and 80% of lanreotide remained intact when incorporated in Lan/Deo-SN2 and Lan/Deo-SN3, respectively. Furthermore, Lan/Deo-SN2 and Lan/Deo-SN3 showed 47% and 51% protection against thiol enriched casein peptones, respectively. Both formulations showed sustained lanreotide release over a period of 3 h. CONCLUSION: Owing to the results, the above-mentioned approach might be a useful tool to overcome the sulfhydryl barrier of the GI-tract.
Assuntos
Peptídeos Cíclicos/administração & dosagem , Peptídeos/administração & dosagem , Somatostatina/análogos & derivados , Tensoativos/química , Dissulfetos/química , Emulsões , Glutationa/química , Humanos , Oxirredução , Peptídeos/química , Peptídeos Cíclicos/química , Somatostatina/administração & dosagem , Somatostatina/química , Compostos de Sulfidrila/químicaRESUMO
The aim of this study was to generate and characterize a chondroitin sulfate-cysteine conjugate (CS-cys) as a novel bioadhesive agent for intra-articular use. Mucoadhesive properties of synthesized CS-cys were investigated by rheological measurement of polymer-mucus mixture and rotating cylinder method, while bioadhesive features of CS-cys on porcine articular cartilage were evaluated via tensile studies. Thiolation was achieved by attachment of l-cysteine to CS via amide bond formation mediated by carbodiimide as a coupling reagent. The conjugate exhibited 421.17±35.14 µmol free thiol groups per gram polymer. The reduced CS-cys displayed 675.09±39.67 µmol free thiol groups per gram polymer after disulfide bonds reduction using tris(2-carboxyethyl)phosphine hydrochloride. The increase in dynamic viscosity of thiolated CS due to oxidative disulfide bond formation was demonstrated using capillary viscometer. The combination of CS-cys and mucus led to 4.57-fold increase in dynamic viscosity in comparison with mucus control. Furthermore, adhesion time to porcine mucosa of CS-cys-based test disk was enhanced by 2.48-fold compared to unmodified CS as measured by rotating cylinder method suggesting the interaction between thiomers and mucus gel layer via disulfide bonds formation. Tensile studies of thiolated CS on porcine articular cartilage showed 5.37- and 1.76-fold increase in the total work of adhesion and the maximum detachment force, respectively, in comparison with unmodified CS indicating bioadhesive features of CS-cys. Cytotoxicity of CS-cys was assessed in Caco-2 cells and rat primary articular chondrocytes using MTT and LDH release assay, thereby showing the safety of CS-cys at a concentration of 0.25% (w/v) in Caco-2 cells. Furthermore, 0.1% of CS-cys was found non-toxic to rat primary articular chondrocytes. According to these results, CS-cys provides improved bioadhesive properties that might be useful as an intra-articular agent for treatment of osteoarthritis.
Assuntos
Adesivos/química , Cartilagem Articular/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Sulfatos de Condroitina/administração & dosagem , Sulfatos de Condroitina/química , Cisteína/química , Polímeros/química , Adesivos/administração & dosagem , Animais , Células CACO-2 , Linhagem Celular Tumoral , Cisteína/administração & dosagem , Dissulfetos/química , Sistemas de Liberação de Medicamentos/métodos , Humanos , Masculino , Mucosa/efeitos dos fármacos , Oxirredução , Fosfinas/química , Polímeros/administração & dosagem , Ratos , Ratos Sprague-Dawley , Reologia/métodos , Compostos de Sulfidrila/química , Suínos , ViscosidadeRESUMO
The objective was to investigate whether even low-molecular weight polymers (LMWPs) can be rendered mucoadhesive due to thiolation. Interceded by the double catalytic system carbodiimide/N-hydroxysuccinimide, cysteamine was covalently attached to a copolymer, poly(4-styrenesulfonic acid-co-maleic acid) (PSSA-MA) exhibiting a molecular weight of just 20 kDa. Depending on the amount of added N-hydroxysuccinimide and cysteamine, the resulting PSSA-MA-cysteamine (PC) conjugates exhibited increasing degree of thiolation, highest being "PC 2300" exhibiting 2300.16 ± 149.86 µmol thiol groups per gram of polymer (mean ± SD; n = 3). This newly developed thiolated polymer was evaluated regarding mucoadhesive, rheological and drug release properties as well from the toxicological point of view. Swelling behavior in 100 mM phosphate buffer pH 6.8 was improved up to 180-fold. Furthermore, due to thiolation, the mucoadhesive properties of the polymer were 240-fold improved. Rheological measurements of polymer/mucus mixtures confirmed results obtained by mucoadhesion studies. In comparison to unmodified polymer, PC 2300 showed 2.3-, 2.3- and 2.4-fold increase in dynamic viscosity, elastic modulus and viscous modulus, respectively. Sustained release of the model drug codeine HCl out of the thiomer was provided for 2.5 h (p < 0.05), whereas the drug was immediately released from the unmodified polymer. Moreover, the thiomer was found non-toxic over Caco-2 cells for a period of 6- and 24-h exposure. Findings of the present study provide evidence that due to thiolation LMWPs can be rendered highly mucoadhesive as well as cohesive and that a controlled drug release out of such polymers can be provided.
Assuntos
Adesivos/química , Polímeros/química , Compostos de Sulfidrila/química , Animais , Células CACO-2 , Carbodi-Imidas/química , Linhagem Celular Tumoral , Cisteamina/química , Sistemas de Liberação de Medicamentos , Humanos , Mucosa Intestinal/metabolismo , Maleatos/química , Peso Molecular , Poliestirenos/química , Reologia , Succinimidas/química , Suínos , ViscosidadeRESUMO
The objective of the present study was to synthesize and characterize cysteamine conjugated ß-cyclodextrin (ß-CD-Cys) as a novel mucoadhesive oligomeric excipient for intra-oral drug delivery. ß-CD-Cys conjugates were obtained in a two-step synthetic pathway, whereby, vicinal diol groups of the oligomer were oxidized using increasing concentrations of sodium-per-iodate (NaIO4), prior to the covalent coupling of cysteamine via reductive amination. Quantification of immobilized thiol groups through Ellman's test revealed 561.56 ± 81 µmol/g, 1054.26 ± 131 µmol/g and 1783.92 ± 201 µmol/g of free thiol groups attached to the oligomer backbone depending on the extent of oxidation. ß-CD-Cys conjugates at concentrations of 0.5% (m/v) showed no toxic effects on Caco-2 cells within 72 h. Furthermore, ß-CD-Cys conjugates displayed a 4-fold improved water solubility compared to the parent oligomer. ß-CD-Cys conjugates (ß-CD-Cys561, ß-CD-Cys1054 and ß-CD-Cys1783) showed 2.86-, 15.09- and 49.08-fold improved retention time on porcine intestinal mucosa and 9.66-, 16.43- and 34.51-fold improved on the porcine buccal mucosa, respectively. Formation of inclusion complexes of miconazole nitrate and ß-CD-Cys1054 resulted in 150-fold increased solubility of miconazole nitrate. According to these results, it seems that ß-CD-Cys conjugates might provide a new promising tool for delivery of poorly water soluble therapeutic agents, such as miconazole nitrate for intra-oral delivery.
Assuntos
Antifúngicos/administração & dosagem , Cisteamina/química , Portadores de Fármacos/química , Miconazol/administração & dosagem , Compostos de Sulfidrila/química , beta-Ciclodextrinas/química , Adesivos/síntese química , Adesivos/química , Administração Oral , Animais , Células CACO-2 , Cisteamina/síntese química , Portadores de Fármacos/síntese química , Sistemas de Liberação de Medicamentos , Humanos , Mucosa Intestinal/metabolismo , Mucosa Bucal/metabolismo , Solubilidade , Compostos de Sulfidrila/síntese química , Suínos , beta-Ciclodextrinas/síntese químicaRESUMO
BACKGROUND: Oral squamous cell carcinoma (OSCC) is mainly caused by smoking and alcohol abuse and shows a five-year survival rate of ~50%. We aimed to explore the variation of somatic mitochondrial DNA (mtDNA) mutations in primary oral tumors, recurrences and metastases. METHODS: We performed an in-depth validation of mtDNA next-generation sequencing (NGS) on an Illumina HiSeq 2500 platform for its application to cancer tissues, with the goal to detect low-level heteroplasmies and to avoid artifacts. Therefore we genotyped the mitochondrial genome (16.6 kb) from 85 tissue samples (tumors, recurrences, resection edges, metastases and blood) collected from 28 prospectively recruited OSCC patients applying both Sanger sequencing and high-coverage NGS (~35,000 reads per base). RESULTS: We observed a strong correlation between Sanger sequencing and NGS in estimating the mixture ratio of heteroplasmies (r = 0.99; p<0.001). Non-synonymous heteroplasmic variants were enriched among cancerous tissues. The proportions of somatic and inherited variants in a given gene region were strongly correlated (r = 0.85; p<0.001). Half of the patients shared mutations between benign and cancerous tissue samples. Low level heteroplasmies (<10%) were more frequent in benign samples compared to tumor samples, where heteroplasmies >10% were predominant. Four out of six patients who developed a local tumor recurrence showed mutations in the recurrence that had also been observed in the primary tumor. Three out of five patients, who had tumor metastases in the lymph nodes of their necks, shared mtDNA mutations between primary tumors and lymph node metastases. The percentage of mutation heteroplasmy increased from the primary tumor to lymph node metastases. CONCLUSIONS: We conclude that Sanger sequencing is valid for heteroplasmy quantification for heteroplasmies ≥10% and that NGS is capable of reliably detecting and quantifying heteroplasmies down to the 1%-level. The finding of shared mutations between primary tumors, recurrences and metastasis indicates a clonal origin of malignant cells in oral cancer.
Assuntos
Carcinoma de Células Escamosas/genética , Genoma Mitocondrial , Sequenciamento de Nucleotídeos em Larga Escala , Neoplasias Bucais/genética , Mutação , Carcinoma de Células Escamosas/patologia , DNA Mitocondrial/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Dados de Sequência Molecular , Neoplasias Bucais/patologia , Recidiva Local de Neoplasia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Análise de Sequência de DNA/métodosRESUMO
In this study we report the novel polymeric resin poly(N-vinyl imidazole/ethylene glycol dimethacrylate) for the purification and isolation of phenolic acids. The monomer to crosslinker ratio and the porogen composition were optimized for isolating phenolic acids diluted in acetonitrile at normal phase chromatography conditions, first. Acetonitrile serves as polar, aprotic solvent, dissolving phenolic acids but not interrupting interactions with the stationary phase due to the approved Hansen solubility parameters. The optimized resin demonstrated high loading capacities and adsorption abilities particularly for phenolic acids in both, acetonitrile and aqueous solutions. The adsorption behavior of aqueous standards can be attributed to ion exchange effects due to electrostatic interactions between protonated imidazole residues and deprotonated phenolic acids. Furthermore, adsorption experiments and subsequent curve fittings provide information of maximum loading capacities of single standards according to the Langmuir adsorption model. Recovery studies of the optimized polymer in the normal-phase and ion-exchange mode illustrate the powerful isolation properties for phenolic acids and are comparable or even better than typical, commercially available solid phase extraction materials. In order to prove the applicability, a highly complex extract of rosemary leaves was purified by poly(N-vinyl imidazole/ethylene glycol dimethacrylate) and the isolated compounds were identified using UHPLC-qTOF-MS.
Assuntos
Hidroxibenzoatos/isolamento & purificação , Imidazóis/química , Metacrilatos/química , Extratos Vegetais/química , Polivinil/química , Rosmarinus/química , Adsorção , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas , Extração em Fase Sólida/métodosRESUMO
The aim of this study was to generate and characterize a thiolated carrageenan. Thiolated carrageenan (carrageenan-SH) was synthesized from kappa (κ)- and iota (ι)-carrageenan by bromine replacement of the hydroxyl moieties followed by substitution to thiol groups using thiourea. Thiolated κ- and ι-carrageenan exhibited 176.57 ± 20.11 and 109.51 ± 18.26 µmol thiol groups per gram polymer, respectively. The resazurin test in Caco-2 cells revealed no toxic effect of both thiolated carrageenans at a concentration below 0.1% (w/v). Regarding efflux pump inhibitory effect, cellular accumulation of multidrug-resistance protein 2 substrate, sulforhodamine 101, was 1.38- and 1.35-fold increased in cells treated with thiolated κ- and ι-carrageenan, respectively. Modification of κ- and ι-carrageenan led to 3.9- and 2.0-fold increase in dynamic viscosity of mucus-thiolated carrageenan mixture within 4 h. Furthermore, residence time of κ- and ι-carrageenan-SH on porcine intestinal mucosa was 6.4- and 1.8-fold prolonged, respectively, as demonstrated by rotating cylinder method, indicating improved mucoadhesive properties. Hence, thiolation of carrageenans led to novel pharmaceutical excipients for various applications.
Assuntos
Antineoplásicos/farmacologia , Antivirais/farmacologia , Carragenina/farmacologia , Fármacos Gastrointestinais/farmacologia , Absorção Intestinal/efeitos dos fármacos , Mucosa Intestinal/efeitos dos fármacos , Compostos de Sulfidrila/farmacologia , Animais , Antineoplásicos/efeitos adversos , Antineoplásicos/química , Antivirais/efeitos adversos , Antivirais/química , Células CACO-2 , Carragenina/efeitos adversos , Carragenina/química , Sobrevivência Celular/efeitos dos fármacos , Enterócitos/efeitos dos fármacos , Enterócitos/metabolismo , Fármacos Gastrointestinais/efeitos adversos , Fármacos Gastrointestinais/química , Humanos , Indicadores e Reagentes/química , Mucosa Intestinal/metabolismo , Moduladores de Transporte de Membrana/efeitos adversos , Moduladores de Transporte de Membrana/química , Moduladores de Transporte de Membrana/farmacologia , Peso Molecular , Proteína 2 Associada à Farmacorresistência Múltipla , Proteínas Associadas à Resistência a Múltiplos Medicamentos/antagonistas & inibidores , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Alga Marinha/química , Compostos de Sulfidrila/efeitos adversos , Compostos de Sulfidrila/química , Sus scrofa , Tioureia/química , ViscosidadeRESUMO
Metal oxides show high selectivity and sensitivity toward mass spectrometry based enrichment strategies. Phosphopeptides/phosphoproteins enrichment from biological samples is cumbersome because of their low abundance. Phosphopeptides are of interest in enzymes and phosphorylation pathways which lead to the clinical links of a disease. Magnetic core-shell lanthanide oxide nanoparticles (Fe3O4@SiO2-La2O3 and Fe3O4@SiO2-Sm2O3) are fabricated, characterized by SEM, FTIR, and EDX and employed in the enrichment of phosphopeptides. The nanoparticles enrich phosphopeptides from casein variants, nonfat milk, egg yolk, human serum and HeLa cell extract. The materials and enrichment protocols are designed in a way that there are almost no nonspecific bindings. The selectivity is achieved up to 1:8500 using ß-casein/BSA mixture and sensitivity down to 1 atto-mole. Batch-to-batch reproducibility is high with the reuse of core-shell nanoparticles up to four cycles. The enrichment followed by MALDI-MS analyses is carried out for the identification of phosphopeptides from serum digest and HeLa cell extract. Characteristic phosphopeptides of phosphoproteins are identified from human serum after the enrichment, which have the diagnostic potential toward prostate cancer. Thus, the lanthanide based magnetic core-shell materials offer a highly selective and sensitive workflow in phosphoproteomics.
Assuntos
Elementos da Série dos Lantanídeos/química , Imãs/química , Nanopartículas/química , Óxidos/química , Fosfoproteínas/análise , Proteoma/análise , Proteômica , Animais , Gema de Ovo/química , Células HeLa , Humanos , Fenômenos Magnéticos , Leite/química , Estrutura Molecular , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Photothermal therapy (PPT) is a platform to fight cancer by using multiplexed interactive plasmonic nanomaterials as probes in combination with the excellent therapeutic performance of near-infrared (NIR) light. With recent rapid developments in optics and nanotechnology, plasmonic materials have potential in cancer diagnosis and treatment, but there are some concerns regarding their clinical use. The primary concerns include the design of plasmonic nanomaterials which are taken up by the tissues, perform their function and then clear out from the body. Gold nanoparticles (Au NPs) can be developed in different morphologies and functionalized to assist the photothermal therapy in a way that they have clinical value. This review outlines the diverse Au morphologies, their distinctive characteristics, concerns and limitations to provide an idea of the requirements in the field of NIR-based therapeutics.