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1.
Environ Sci Technol ; 58(36): 15949-15959, 2024 Sep 10.
Artículo en Inglés | MEDLINE | ID: mdl-39143808

RESUMEN

Fluorinated liquid-crystal monomers (FLCMs) are a potential emerging class of persistent, bioaccumulative, and toxic compounds. Humans inevitably ingest FLCMs via food and the environment. However, there are limited studies on internal exposure biomonitoring of FLCMs. Herein, we evaluated the estimated daily intakes (EDIs) of FLCMs in the general population based on serum residue levels. For the first time, 38 FLCMs were detected in 314 serum samples from the general population in Beijing, with a median value of 132.48 ng/g of lipid weight (lw). BDPrB is a predominant FLCM in serum. The median EDI of ∑38FLCMs in the general residents was 37.96 pg/kg bw/day. The residual levels of most FLCMs were higher in urban than in suburban areas (p < 0.05). The concentrations of EFPEB, EDPrB, EDFPBB, and PDTFMTFT in serum showed positive associations with blood glucose (GLU) (r = 0.126-0.275, p < 0.05). Logistic regression analysis showed that FLCMs were significantly positively correlated with dyslipidemia, with an odds ratio of 2.19; BDPrB was significantly positively correlated with hyperglycemia (OR: 2.48). Overall, the present study suggests the occurrence of FLCMs in the nonoccupational population, and the exposure of certain FLCMs may cause abnormal blood glucose and lipid levels.


Asunto(s)
Cristales Líquidos , Suero , Femenino , Humanos , Masculino , Cristales Líquidos/análisis , Suero/química
2.
Int J Mol Sci ; 25(13)2024 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-39000368

RESUMEN

Selenium is an essential trace element in our diet, crucial for the composition of human selenoproteins, which include 25 genes such as glutathione peroxidases and thioredoxin reductases. The regulation of the selenoproteome primarily hinges on the bioavailability of selenium, either from dietary sources or cell culture media. This selenium-dependent control follows a specific hierarchy, with "housekeeping" selenoproteins maintaining constant expression while "stress-regulated" counterparts respond to selenium level fluctuations. This study investigates the variability in fetal bovine serum (FBS) selenium concentrations among commercial batches and its effects on the expression of specific stress-related cellular selenoproteins. Despite the limitations of our study, which exclusively used HEK293 cells and focused on a subset of selenoproteins, our findings highlight the substantial impact of serum selenium levels on selenoprotein expression, particularly for GPX1 and GPX4. The luciferase reporter assay emerged as a sensitive and precise method for evaluating selenium levels in cell culture environments. While not exhaustive, this analysis provides valuable insights into selenium-mediated selenoprotein regulation, emphasizing the importance of serum composition in cellular responses and offering guidance for researchers in the selenoprotein field.


Asunto(s)
Selenio , Selenoproteínas , Selenio/sangre , Selenio/metabolismo , Humanos , Selenoproteínas/genética , Selenoproteínas/metabolismo , Bovinos , Animales , Células HEK293 , Glutatión Peroxidasa/metabolismo , Glutatión Peroxidasa/genética , Glutatión Peroxidasa GPX1 , Suero/metabolismo , Suero/química , Fosfolípido Hidroperóxido Glutatión Peroxidasa/metabolismo , Fosfolípido Hidroperóxido Glutatión Peroxidasa/genética , Medios de Cultivo/química , Regulación de la Expresión Génica/efectos de los fármacos
3.
Clin Chim Acta ; 562: 119851, 2024 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-38977172

RESUMEN

BACKGROUND: Observable quantitative variations exist between plasma and serum in routine protein measurements, often not reflected in standard reference intervals. In this study, we describe an indirect approach for estimating a combined reference interval (RI) (i.e., serum and plasma), for commonly ordered protein measurands: total protein, albumin, and globulin. METHODS: We applied an indirect reference interval estimation for protein measurements in serum and plasma using data from July 2018 to February 2024. The data were divided into three Epochs based on a period of plasma separator tube shortage during the COVID-19 pandemic. Bootstrap resampling was used to calculate RIs and corresponding 95% confidence intervals for each month. RESULTS: Our results demonstrate notable changes in RI limits for total protein, albumin, and globulin between Epochs, reflecting the influence of changing sample matrix. A combined RI was identified for all components and verified using plasma and serum samples from 20 healthy individuals and retrospective analysis of flagging rates on our outpatient population using new and historical RIs. CONCLUSION: The study demonstrates notable differences in the RIs for total protein, albumin, and globulin when container type changes. In addition, the results demonstrate the effectiveness of big data analytics in deriving RIs and highlights the necessity of continuous RI assessment and adjustment based on the patient population and acceptable specimen types.


Asunto(s)
Globulinas , Albúmina Sérica , Humanos , Valores de Referencia , Globulinas/análisis , Albúmina Sérica/análisis , COVID-19/sangre , Estudios Retrospectivos , Proteínas Sanguíneas/análisis , Masculino , Plasma/química , Femenino , Adulto , Persona de Mediana Edad , Suero/química , SARS-CoV-2 , Seroglobulinas/análisis , Análisis Químico de la Sangre/normas , Análisis Químico de la Sangre/métodos
4.
Eur J Cell Biol ; 103(3): 151445, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-39024989

RESUMEN

Allogeneic serum and tissue-specific extracellular matrix have been shown to maintain permanently differentiated cell phenotype in culture. This is of particular importance for human tenocytes, a cell population that readily loses its function during ex vivo culture. With these in mind, herein we extracted human tenocytes using either foetal bovine serum or human serum, cultured them in the absence and presence of carrageenan and Ficoll®, the most widely used macromolecular crowding agents (to induce tissue-specific extracellular matrix deposition), and assessed cellular function, via metabolic activity, viability, proliferation and immunofluorescence for collagen related molecules, non-collagenous molecules and transmembrane molecules. At day 7, longest time point assessed, neither carrageenan nor Ficoll® significantly affected metabolic activity, viability and proliferation in either serum and human serum significantly increased metabolic activity and proliferation. At day 7, in the absence of macromolecular crowding, cells in human serum deposited significantly lower collagen type VI, biglycan, versican and tenomodulin than cells in foetal bovine serum. Interestingly, at day 7, in comparison to the no macromolecular crowding group, carrageenan in foetal bovine serum induced the highest effect, as judged by the highest number of significantly increased molecules (collagen type I, collagen type IV, collagen type V, collagen type VI, transforming growth factor ß1, matrix metalloproteinase 14, lumican, versican, scleraxis and integrin α2ß1). These data, although contradict previous observations where human serum outperformed foetal bovine serum, at the same time, support the use of foetal bovine serum in the development of cell-based medicines.


Asunto(s)
Tenocitos , Humanos , Tenocitos/metabolismo , Tenocitos/citología , Células Cultivadas , Proliferación Celular , Animales , Suero/metabolismo , Suero/química , Bovinos , Carragenina/farmacología , Ficoll , Matriz Extracelular/metabolismo
5.
Clin Biochem ; 131-132: 110804, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39079578

RESUMEN

INTRODUCTION: Serum is the International Federation of Clinical Chemistry (IFCC)-recommended matrix for the measurement of lactate dehydrogenase (LD); however, many laboratories opt for lithium heparin plasma to achieve quicker turnaround times and minimize tube usage. When introducing the new Sigma-Strong IFCC-recommended LDH2 assay from Abbott Laboratories on lithium-heparin collected samples, we observed a rise in the patient median LD activity as well as several samples exhibiting falsely elevated values. MATERIALS AND METHODS: 120 + serum and plasma samples from consenting patients were collected and evaluated for complete blood count and lactate dehydrogenase using two different assays. Aggregated patient results before and after introduction of the LDH2 assay were compared. RESULTS: Mean LD was 14% higher in plasma than in serum when using the LDH2 assay but only 5% higher when using the previous LDH legacy assay from Abbott Laboratories. Similarly, platelets and leukocytes were 10-30 times higher in plasma than in serum. Aggregated lactate dehydrogenase patient results demonstrated a dramatic increase in patient median following introduction of the LDH2 assay. Various experiments were tried to reduce cellular interference, but the only viable solution we found, apart from reverting to the LDH legacy assay, was to utilize serum tubes. CONCLUSION: We conclude that lithium-heparin plasma leads to falsely elevated lactate dehydrogenase activity when using the LDH2 assay. These errors can be prevented by using serum collected in gel separator tubes.


Asunto(s)
L-Lactato Deshidrogenasa , Humanos , L-Lactato Deshidrogenasa/sangre , Plasma/química , Heparina/sangre , Suero/química , Reacciones Falso Positivas
6.
Parasite Immunol ; 46(7): e13058, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-39072810

RESUMEN

Neurocysticercosis (NCC), a major cause of global acquired epilepsy, results from Taenia solium larval brain infection. T. solium adult worms release large numbers of infective eggs into the environment contributing to high levels of exposure in endemic areas. This study identifies T. solium proteins in the sera of individuals with and without NCC using mass spectrometry to examine exposure in endemic regions. Forty-seven patients (18-51 years), 24 parenchymal NCC (pNCC), 8 epilepsy of unknown aetiology, 7 glioma, 8 brain tuberculoma, and 7 healthy volunteers were studied. Trypsin digested sera were subject to liquid chromatography-tandem mass spectrometry and spectra of 375-1700 m/z matched against T. solium WormBase ParaSite database with MaxQuant software to identify T. solium proteins. Three hundred and nineteen T. solium proteins were identified in 87.5% of pNCC and 56.6% of non-NCC subjects. Three hundred and four proteins were exclusive to pNCC sera, seven to non-NCC sera and eight in both. Ten percent, exhibiting immune-modulatory properties, originated from the oncosphere and cyst vesicular fluid. In conclusion, in endemic regions, T. solium proteins are detected in sera of individuals with and without pNCC. The immunomodulatory nature of these proteins may influence susceptibility and course of infection.


Asunto(s)
Proteínas del Helminto , Neurocisticercosis , Taenia solium , Humanos , Neurocisticercosis/sangre , Neurocisticercosis/parasitología , Taenia solium/inmunología , Adulto , Adolescente , Animales , Persona de Mediana Edad , Adulto Joven , Masculino , Femenino , Proteínas del Helminto/sangre , Cromatografía Liquida , Espectrometría de Masas en Tándem , Espectrometría de Masas , Suero/química
7.
Clin Exp Med ; 24(1): 133, 2024 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-38900301

RESUMEN

This study aimed to investigate the serum and expression levels of C-X-C motif chemokine ligand 9 (CXCL9), CXCL10, CXCL11, and CXC receptor 3 (CXCR3) in minor salivary glands (MSGs) of patients with primary Sjögren's syndrome (pSS), and to explore their correlations with clinical parameters. Serum samples from 49 patients diagnosed with pSS, 33 patients with rheumatoid arthritis (RA), and 30 healthy controls (HCs) were collected for measurements of CXCL9, CXCL10, CXCL11, and CXCR3. Additionally, CXCL levels in the MSG tissues were measured in 41 patients who underwent MSG biopsy. Correlations between CXCL and CXCL/CXCR levels in serum/MSG tissues and clinical factors/salivary scintigraphy parameters were analyzed. Serum CXCL11 and CXCR3 showed statistically significant differences among patients with pSS and RA and HCs (serum CXCL11, pSS:RA:HC = 235.6 ± 500.1 pg/mL:90.0 ± 200.3 pg/mL:45.9 ± 53.6 pg/mL; p = 0.041, serum CXCR3, pSS:RA:HC = 3.27 ± 1.32 ng/mL:3.29 ± 1.17 ng/mL:2.00 ± 1.12 ng/mL; p < 0.001). Serum CXCL10 showed a statistically significant difference between pSS (64.5 ± 54.2 pg/mL) and HCs (18.6 ± 18.1 pg/mL, p < 0.001), while serum CXCL9 did not exhibit a significant difference among the groups. Correlation analysis of clinical factors revealed that serum CXCL10 and CXCL11 levels positively correlated with erythrocyte sedimentation rate (r = 0.524, p < 0.001 and r = 0.707, p < 0.001, respectively), total protein (r = 0.375, p = 0.008 and r = 0.535, p < 0.001, respectively), globulin (r = 0.539, p < 0.001 and r = 0.639, p < 0.001, respectively), and European Alliance of Associations for Rheumatology SS Disease Activity Index (r = 0.305, p = 0.033 and r = 0.321, p = 0.025). Additionally, serum CXCL10 negatively correlated with the Schirmer test score (r = - 0.354, p = 0.05), while serum CXCL11 positively correlated with the biopsy focus score (r = 0.612, p = 0.02). In the MSG tissue, the percentage of infiltrating CXCL9-positive cells was highest (75.5%), followed by CXCL10 (29.1%) and CXCL11 (27.9%). In the correlation analysis, CXCL11-expressing cells were inversely related to the mean washout percentage on salivary gland scintigraphy (r = - 0.448, p = 0.007). Our study highlights distinct serum and tissue chemokine patterns in pSS, emphasizing CXCL9's potential for early diagnosis. This suggests that CXCL10 and CXCL11 are indicators of disease progression, warranting further investigation into their roles in autoimmune disorders beyond pSS.


Asunto(s)
Quimiocina CXCL10 , Quimiocina CXCL11 , Receptores CXCR3 , Síndrome de Sjögren , Humanos , Síndrome de Sjögren/patología , Síndrome de Sjögren/sangre , Síndrome de Sjögren/metabolismo , Femenino , Persona de Mediana Edad , Masculino , Receptores CXCR3/metabolismo , Adulto , Quimiocina CXCL11/sangre , Quimiocina CXCL10/sangre , Anciano , Glándulas Salivales Menores/patología , Glándulas Salivales Menores/metabolismo , Quimiocina CXCL9/sangre , Suero/química , Suero/metabolismo
8.
Biomolecules ; 14(6)2024 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-38927049

RESUMEN

We recently reported the potential application of recombinant prothrombin activator ecarin (RAPClot™) in blood diagnostics. In a new study, we describe RAPClot™ as an additive to develop a novel blood collection prototype tube that produces the highest quality serum for accurate biochemical analyte determination. The drying process of the RAPClot™ tube generated minimal effect on the enzymatic activity of the prothrombin activator. According to the bioassays of thrombin activity and plasma clotting, γ-radiation (>25 kGy) resulted in a 30-40% loss of the enzymatic activity of the RAPClot™ tubes. However, a visual blood clotting assay revealed that the γ-radiation-sterilized RAPClot™ tubes showed a high capacity for clotting high-dose heparinized blood (8 U/mL) within 5 min. This was confirmed using Thrombelastography (TEG), indicating full clotting efficiency under anticoagulant conditions. The storage of the RAPClot™ tubes at room temperature (RT) for greater than 12 months resulted in the retention of efficient and effective clotting activity for heparinized blood in 342 s. Furthermore, the enzymatic activity of the RAPClot™ tubes sterilized with an electron-beam (EB) was significantly greater than that with γ-radiation. The EB-sterilized RAPClot™ tubes stored at RT for 251 days retained over 70% enzyme activity and clotted the heparinized blood in 340 s after 682 days. Preliminary clinical studies revealed in the two trials that 5 common analytes (K, Glu, lactate dehydrogenase (LD), Fe, and Phos) or 33 analytes determined in the second study in the γ-sterilized RAPClot™ tubes were similar to those in commercial tubes. In conclusion, the findings indicate that the novel RAPClot™ blood collection prototype tube has a significant advantage over current serum or lithium heparin plasma tubes for routine use in measuring biochemical analytes, confirming a promising application of RAPClot™ in clinical medicine.


Asunto(s)
Proteínas Recombinantes , Humanos , Coagulación Sanguínea/efectos de los fármacos , Suero/química , Suero/metabolismo , Tromboplastina/metabolismo , Recolección de Muestras de Sangre/métodos , Tromboelastografía/métodos , Rayos gamma , Anticoagulantes/farmacología , Anticoagulantes/química
9.
Spectrochim Acta A Mol Biomol Spectrosc ; 317: 124390, 2024 Sep 05.
Artículo en Inglés | MEDLINE | ID: mdl-38749203

RESUMEN

Label-free Surface Enhanced Raman Spectroscopy (SERS) is a rapid technique that has been extensively applied in clinical diagnosis and biomedicine for the analysis of biofluids. The purpose of this approach relies on the ability to detect specific "metabolic fingerprints" of complex biological samples, but the full potential of this technique in diagnostics is yet to be exploited, mainly because of the lack of common analytical protocols for sample preparation and analysis. Variation of experimental parameters, such as substrate type, laser wavelength and sample processing can greatly influence spectral patterns, making results from different research groups difficult to compare. This study aims at making a step toward a standardization of the protocols in the analysis of human serum samples with Ag nanoparticles, by directly comparing the SERS spectra obtained from five different methods in which parameters like laser power, nanoparticle concentration, incubation/deproteinization steps and type of substrate used vary. Two protocols are the most used in the literature, and the other three are "in-house" protocols proposed by our group; all of them are employed to analyze the same human serum sample. The experimental results show that all protocols yield spectra that share the same overall spectral pattern, conveying the same biochemical information, but they significantly differ in terms of overall spectral intensity, repeatability, and preparation steps of the sample. A Principal Component Analysis (PCA) was performed revealing that protocol 3 and protocol 1 have the least variability in the dataset, while protocol 2 and 4 are the least repeatable.


Asunto(s)
Nanopartículas del Metal , Análisis de Componente Principal , Plata , Espectrometría Raman , Humanos , Espectrometría Raman/métodos , Nanopartículas del Metal/química , Plata/química , Suero/química
10.
Int J Mol Sci ; 25(10)2024 May 13.
Artículo en Inglés | MEDLINE | ID: mdl-38791352

RESUMEN

Biofunctionalized hydrogels are widely used in tissue engineering for bone repair. This study examines the bone regenerative effect of the blood-derived growth factor preparation of Hypoxia Preconditioned Serum (HPS) and its fibrin-hydrogel formulation (HPS-F) on drilled defects in embryonic day 19 chick femurs. Measurements of bone-related growth factors in HPS reveal significant elevations of Osteopontin, Osteoprotegerin, and soluble-RANKL compared with normal serum (NS) but no detection of BMP-2/7 or Osteocalcin. Growth factor releases from HPS-F are measurable for at least 7 days. Culturing drilled femurs organotypically on a liquid/gas interface with HPS media supplementation for 10 days demonstrates a 34.6% increase in bone volume and a 52.02% increase in bone mineral density (BMD) within the defect area, which are significantly higher than NS and a basal-media-control, as determined by microcomputed tomography. HPS-F-injected femur defects implanted on a chorioallantoic membrane (CAM) for 7 days exhibit an increase in bone mass of 123.5% and an increase in BMD of 215.2%, which are significantly higher than normal-serum-fibrin (NS-F) and no treatment. Histology reveals calcification, proteoglycan, and collagen fiber deposition in the defect area of HPS-F-treated femurs. Therefore, HPS-F may offer a promising and accessible therapeutic approach to accelerating bone regeneration by a single injection into the bone defect site.


Asunto(s)
Regeneración Ósea , Fémur , Fibrina , Animales , Regeneración Ósea/efectos de los fármacos , Fémur/efectos de los fármacos , Fémur/diagnóstico por imagen , Fémur/metabolismo , Fibrina/metabolismo , Embrión de Pollo , Densidad Ósea/efectos de los fármacos , Hidrogeles , Microtomografía por Rayos X , Ingeniería de Tejidos/métodos , Suero/metabolismo , Suero/química
11.
Nanomedicine ; 58: 102745, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38499167

RESUMEN

Understanding the stability of mRNA loaded lipid nanoparticles (mRNA-LNPs) is imperative for their clinical development. Herein, we propose the use of size-exclusion chromatography coupled with dual-angle light scattering (SEC-MALS) as a new approach to assessing mRNA-LNP stability in pure human serum and plasma. By applying a dual-column configuration to attenuate interference from plasma components, SEC-MALS was able to elucidate the degradation kinetics and physical property changes of mRNA-LNPs, which have not been observed accurately by conventional dynamic light scattering techniques. Interestingly, both serum and plasma had significantly different impacts on the molecular weight and radius of gyration of mRNA-LNPs, suggesting the involvement of clotting factors in desorption of lipids from mRNA-LNPs. We also discovered that a trace impurity (~1 %) in ALC-0315, identified as its O-tert-butyloxycarbonyl-protected form, greatly diminished mRNA-LNP stability in serum. These results demonstrated the potential utility of SEC-MALS for optimization and quality control of LNP formulations.


Asunto(s)
Cromatografía en Gel , Lípidos , Nanopartículas , ARN Mensajero , Humanos , ARN Mensajero/genética , ARN Mensajero/sangre , Nanopartículas/química , Lípidos/química , Dispersión Dinámica de Luz , Plasma/química , Luz , Dispersión de Radiación , Suero/química , Estabilidad del ARN , Liposomas
12.
Clin Chem Lab Med ; 62(8): 1557-1569, 2024 Jul 26.
Artículo en Inglés | MEDLINE | ID: mdl-38443327

RESUMEN

OBJECTIVES: The pre-analytical stability of various biochemical analytes requires careful consideration, as it can lead to the release of erroneous laboratory results. There is currently significant variability in the literature regarding the pre-analytical stability of various analytes. The aim of this study was to determine the pre-analytical stability of 65 analytes in whole blood, serum and plasma using a standardized approach. METHODS: Blood samples were collected from 30 healthy volunteers (10 volunteers per analyte) into five vacutainers; either SST, Li-heparin, K2-EDTA, or Na-fluoride/K-oxalate. Several conditions were tested, including delayed centrifugation with storage of whole blood at room temperature (RT) for 8 h, delayed centrifugation with storage of whole blood at RT or 4 °C for 24 h, and immediate centrifugation with storage of plasma or serum at RT for 24 h. Percent deviation (% PD) from baseline was calculated for each analyte and compared to the maximum permissible instability (MPI) derived from intra- and inter-individual biological variation. RESULTS: The majority of the analytes evaluated remained stable across all vacutainer types, temperatures, and timepoints tested. Glucose, potassium, and aspartate aminotransferase, among others, were significantly impacted by delayed centrifugation, having been found to be unstable in whole blood specimens stored at room temperature for 8 h. CONCLUSIONS: The data presented provides insight into the pre-analytical variables that impact the stability of routine biochemical analytes. This study may help to reduce the frequency of erroneous laboratory results released due to exceeded stability and reduce unnecessary repeat phlebotomy for analytes that remain stable despite delayed processing.


Asunto(s)
Recolección de Muestras de Sangre , Plasma , Suero , Humanos , Recolección de Muestras de Sangre/métodos , Plasma/química , Suero/química , Análisis Químico de la Sangre/métodos , Análisis Químico de la Sangre/normas , Adulto , Masculino , Temperatura , Femenino , Voluntarios Sanos , Centrifugación
13.
Sci Total Environ ; 926: 171908, 2024 May 20.
Artículo en Inglés | MEDLINE | ID: mdl-38527533

RESUMEN

The current study examined the level of Polychlorinated biphenyls (PCBs) in tumor and blood serum of female breast cancer patients and control individuals recruited from Punjab, Pakistan. Breast tumor and blood serum from 40 patients and only blood serum from ten control subjects were obtained and concentration of 32 PCB congeners was analyzed through Gas chromatography coupled with Mass spectrophotometry. Sociodemographic variables of the patients along with essential clinical and haematological parameters were taken as covariates. Tumor reflects the highest median (min-max) concentration (ng g-1 lw) of Æ©PCBs at 115.94 (0.05-17.75) followed by 16.53 (0.09-2.94) and 5.24 (0.01-0.59) in blood serum of cancer patients and control group respectively. Median concentrations (ng g-1 lw) of non-dioxine like Æ©PCBs were considerably higher at 83.04, 32.89 and 4.27 compared to 13.03 and 3.50 and 0.97 for dioxin like Æ©PCBs in tumor, serum of breast cancer patients and control subjects respectively. PCB-87 was most dominant congeners in tumor followed by PCB-170 and -82 whereas PCB-28 and -52 reflected greatest contribution in serum of breast cancer patients. Blood haemoglobin, potassium and chloride ions showed significant positive whereas body mass index reflect inverse relationship when regressed with Æ©PCBs in tumor. This pioneer study depicts elevated concentrations of PCBs in patients compared to control, reflecting potential positive association of PCBs with breast cancer which need further confirmation. We concluded that chronic exposure to PCBs might be associated with an increasing number of breast cancer incidences in developing countries like Pakistan, which should be further elucidated through detail in vitro and in vivo studies.


Asunto(s)
Neoplasias de la Mama , Contaminantes Ambientales , Bifenilos Policlorados , Dibenzodioxinas Policloradas , Humanos , Femenino , Bifenilos Policlorados/análisis , Neoplasias de la Mama/epidemiología , Suero/química , Pakistán/epidemiología , Dibenzodioxinas Policloradas/análisis , Contaminantes Ambientales/análisis
14.
J Pept Sci ; 30(8): e3597, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38523558

RESUMEN

The recently developed mRNA-based coronavirus SARS-CoV-2 vaccines highlighted the great therapeutic potential of the mRNA technology. Although the lipid nanoparticles used for the delivery of the mRNA are very efficient, they showed, in some cases, the induction of side effects as well as the production of antibodies directed against particle components. Thus, the development of alternative delivery systems is of great interest in the pursuit of more effective mRNA treatments. In the present work, we evaluated the mRNA transfection capacities of a series of cationic histidine-rich amphipathic peptides derived from LAH4. We found that while the LAH4-A1 peptide was an efficient carrier for mRNA, its activity was highly serum sensitive. Interestingly, modification of this cell penetrating peptide at the N-terminus with two tyrosines or with salicylic acid allowed to confer serum resistance to the carrier.


Asunto(s)
ARN Mensajero , ARN Mensajero/genética , ARN Mensajero/química , Humanos , SARS-CoV-2/química , SARS-CoV-2/genética , SARS-CoV-2/inmunología , Suero/química , Suero/metabolismo , Transfección/métodos , Péptidos de Penetración Celular/química , Péptidos de Penetración Celular/metabolismo , Nanopartículas/química , Péptidos/química , Animales , COVID-19
15.
J Proteome Res ; 23(8): 3064-3075, 2024 Aug 02.
Artículo en Inglés | MEDLINE | ID: mdl-38520676

RESUMEN

Metabolomics is an emerging and powerful bioanalytical method supporting clinical investigations. Serum and plasma are commonly used without rational prioritization. Serum is collected after blood coagulation, a complex biochemical process involving active platelet metabolism. This may affect the metabolome and increase the variance, as platelet counts and function may vary substantially in individuals. A multiomics approach systematically investigating the suitability of serum and plasma for clinical studies demonstrated that metabolites correlated well (n = 461, R2 = 0.991), whereas lipid mediators (n = 83, R2 = 0.906) and proteins (n = 322, R2 = 0.860) differed substantially between specimen. Independently, analysis of platelet releasates identified most biomolecules significantly enriched in serum compared to plasma. A prospective, randomized, controlled parallel group metabolomics trial with acetylsalicylic acid administered for 7 days demonstrated that the apparent drug effects significantly differ depending on the analyzed specimen. Only serum analyses of healthy individuals suggested a significant downregulation of TXB2 and 12-HETE, which were specifically formed during coagulation in vitro. Plasma analyses reliably identified acetylsalicylic acid effects on metabolites and lipids occurring in vivo such as an increase in serotonin, 15-deoxy-PGJ2 and sphingosine-1-phosphate and a decrease in polyunsaturated fatty acids. The present data suggest that plasma should be preferred above serum for clinical metabolomics studies as the serum metabolome may be substantially confounded by platelets.


Asunto(s)
Aspirina , Plaquetas , Metabolómica , Plasma , Humanos , Plaquetas/metabolismo , Plaquetas/efectos de los fármacos , Metabolómica/métodos , Aspirina/farmacología , Plasma/metabolismo , Plasma/química , Suero/metabolismo , Suero/química , Lisofosfolípidos/sangre , Esfingosina/análogos & derivados , Esfingosina/sangre , Metaboloma/efectos de los fármacos , Tromboxano B2/sangre , Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico/sangre , Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico/metabolismo , Masculino , Femenino , Estudios Prospectivos , Adulto
16.
Clin Gastroenterol Hepatol ; 22(9): 1950-1952.e1, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38423347

RESUMEN

Although ammonia is involved in the pathophysiology of hepatic encephalopathy (HE), the use of ammonia levels in clinical practice is problematic.1-3 For example, in a study of 551 patients with overt HE (OHE) receiving lactulose who had ammonia levels tested, only 60% had an increased ammonia level (defined as >72 µmol/L).2 Overall, there was no correlation observed between lactulose dose and whether ammonia levels were obtained (ie, presence/absence of increased ammonia level did not guide therapy), or between time to OHE resolution and ammonia levels.2 Additionally, there is substantial interlaboratory variability in sample handling and processing, which may affect ammonia measurements.4.


Asunto(s)
Amoníaco , Encefalopatía Hepática , Cirrosis Hepática , Humanos , Encefalopatía Hepática/sangre , Encefalopatía Hepática/diagnóstico , Amoníaco/sangre , Cirrosis Hepática/complicaciones , Cirrosis Hepática/sangre , Masculino , Femenino , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Suero/química , Anciano , Hospitalización , Lactulosa
17.
Biopreserv Biobank ; 22(4): 363-372, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38416864

RESUMEN

Recent studies highlight the presence of bacterial sequences in the human blood, suggesting potential clinical significance for circulating microbial signatures. These sequences could presumably serve in the diagnosis, prediction, or monitoring of various health conditions. Ensuring the similarity of samples before bacterial analysis is crucial, especially when combining samples from different biobanks prepared under varying conditions (such as different DNA extraction kits, centrifugation conditions, blood collection tubes, etc.). In this study, we aimed to analyze the impact of different sample collection and nucleic acid extraction criteria (blood collection tube, centrifugation, input volume, and DNA extraction kit) on circulating bacterial composition. Blood samples from four healthy individuals were collected into three different sample collection tubes: K2EDTA plasma tube, sodium citrate plasma tube, and gel tube for blood serum. Tubes were centrifugated at standard and double centrifugation conditions. DNA extraction was performed using 100, 200, and 500 µL plasma/serum input volumes. DNA extraction was performed using three different isolation kits: Norgen plasma/serum cell-free circulating DNA purification micro kit, Applied Biosystems MagMAX cell-free DNA isolation kit, and Qiagen QIAamp MinElute cell-free circulating DNA mini kit. All samples were subjected to 16S rRNA V1-V2 library preparation and sequencing. In total, 216 DNA and 18 water control samples were included in the study. According to PERMANOVA, PCoA, Mann-Whitney, and FDR tests the effect of the DNA extraction kit on the microbiota composition was the greatest, whereas the type of blood collection tube, centrifugation type, and sample input volume for the extraction had minor effects. Samples extracted with the Norgen DNA extraction kit were enriched with Gram-negative bacteria, whereas samples extracted with the Qiagen and MagMAX kits were enriched with Gram-positive bacteria. Bacterial profiles of samples prepared with the Qiagen and MagMAX DNA extraction kits were more similar, whereas samples prepared with the Norgen DNA extraction kit were significantly different from other groups.


Asunto(s)
Bancos de Muestras Biológicas , Ácidos Nucleicos Libres de Células , ADN Bacteriano , ARN Ribosómico 16S , Humanos , ARN Ribosómico 16S/genética , ADN Bacteriano/aislamiento & purificación , ADN Bacteriano/genética , ADN Bacteriano/sangre , Ácidos Nucleicos Libres de Células/sangre , Ácidos Nucleicos Libres de Células/aislamiento & purificación , Plasma/química , Plasma/microbiología , Suero/química , Suero/microbiología , Bacterias/genética , Bacterias/aislamiento & purificación , Manejo de Especímenes/métodos , Recolección de Muestras de Sangre/métodos , Análisis de Secuencia de ADN/métodos
18.
Anal Methods ; 16(10): 1473-1479, 2024 03 07.
Artículo en Inglés | MEDLINE | ID: mdl-38404261

RESUMEN

A label-free electrochemical immunosensor utilising nitrogen-rich mesoporous carbon (MNC) as the substrate material was developed for the sensitive quantification of carcinoembryonic antigen (CEA). The synergic interactions between MNC and AbCEA also eliminated the need for coupling agents such as EDC/NHS. The novel immunosensor demonstrated a wide detection range from 500 fM (9.04 pg mL-1) to 50 nM (1 µg mL-1) and a low detection limit (LOD) of 500 fM. Moreover, the immunosensor showed sensitivities of 12.27 mA nM-1 cm-2 and 0.066 mA nM-1 cm-2 for detecting CEA in the linear ranges 10 pM to 1 nM and 2 nM to 50 nM, respectively, while maintaining long-term storage stability of 6 weeks. Analysis of real serum sample analysis yielded highly accurate results with recovery rates ranging from 99.3% to 103.7%. Furthermore, the developed paper-based screen-printed electrode exhibited a similar detection range, suggesting its potential for use in point-of-care detection devices in future applications.


Asunto(s)
Técnicas Biosensibles , Antígeno Carcinoembrionario , Antígeno Carcinoembrionario/análisis , Técnicas Biosensibles/métodos , Suero/química , Técnicas Electroquímicas , Inmunoensayo/métodos
19.
Environ Int ; 183: 108440, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38232504

RESUMEN

BACKGROUND: Polychlorinated alkanes (PCAs) constitute a large group of individual congeners originating from commercial chlorinated paraffin (CP) products with carbon chain lengths of PCAs-C10-13, PCAs-C14-17, and PCAs-C18-32, occasionally containing PCAs-C6-9 impurities. The extensive use of CPs has led to global environmental pollution of PCAs. This study aimed to quantify PCAs in paired serum and breast milk of lactating Swedish mothers, exploring their concentration relationship. METHODS: Twenty-five paired samples of mothers' blood serum and breast milk were analysed and concentrations were determined for PCAs C6-32 and compared to 4,4'-DDE, the PCB congener 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153), and hexachlorobenzene (HCB). RESULTS: The median concentrations of PCAs-C6-9, PCAs-C10-13, PCAs-C14-17, PCAs-C18-32 and ΣPCAs in serum were 14, 790, 520, 16 and 1350 ng/g lipid weight (lw), respectively, and in breast milk 0.84, 36, 63, 6.0 and 107 ng/g lw. Levels of 4,4'-DDE, CB-153 and HCB were comparable in the two matrices, serum and breast milk at 17, 12 and 4.9 ng/g lw. The results show significant differences of PCAs-C10-13 and PCAs-C14-17 in breast milk with 22- and 6.2-times lower lw-based concentrations than those measured in serum. On wet weight the differences serum/breast milk ratios of PCAs-C6-9, PCAs-C10-13, PCAs-C14-17, PCAs-C18-32 and ΣPCAs were 1.7, 3.2, 1.0, 0.4 and 1.6, respectively, while the ratio for 4,4'-DDE, CB-153 and HCB were each close to 0.1. CONCLUSION: Swedish lactating mothers had high serum concentrations of PCAs-C10-13 and PCAs-C14-17, with the ΣPCAs median serum concentration of 1350 ng/g lw. The breast milk concentration, although considerably lower at 107 ng/g lw, still surpassed those of 4,4'-DDE, CB-153 and HCB, suggesting an exposure risk of infants to PCAs. The variation in blood and breast milk accumulation between PCAs and studied legacy POPs, is rarely discussed but warrants further studies on partitioning properties as well as associated toxicological implications.


Asunto(s)
Contaminantes Ambientales , Hidrocarburos Clorados , Bifenilos Policlorados , Femenino , Lactante , Humanos , Leche Humana/química , Lactancia , Hexaclorobenceno , Suero/química , Suecia , Estudios de Cohortes , Hidrocarburos Clorados/análisis
20.
Toxicol Ind Health ; 40(3): 134-143, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38289205

RESUMEN

Mercury is a ubiquitous environmental xenobiotic; the primary sources of exposure to this metal are artisanal gold mining and the direct production of mercury. In Mexico, artisanal mercury mining continues to be an important activity in different regions of the country. Exposure to mercury vapors releases can have severe health impacts, including immunotoxic effects such as alterations in cytokine profiling. Therefore, in the present work, we evaluated the inflammatory cytokines profile in the blood serum of miners exposed to mercury. A cross-sectional observational study was performed on 27 mining workers (exposed group) and 20 control subjects (nonexposed group) from central Mexico. The mercury urine concentration (U-Hg) was determined by atomic absorption spectrometry, and IL-2, IL-6, IL-8, IL-10, and TNF-α were measured using a Multiplex Assay. The results showed that the U-Hg in the miners had a median value of 552.70 µg/g creatinine. All cytokines showed a significant increase in the miner group compared with the control group, except for TNF-α. In addition, we observed a positive correlation between U-Hg concentration and cytokine levels. In conclusion, mercury exposure correlated with cytokine levels (considered acute inflammatory marker) in miners; therefore, workers exposed to this metal show an acute systemic inflammation that could lead to alterations in other organs and systems.


Asunto(s)
Mercurio , Exposición Profesional , Humanos , Mercurio/análisis , Exposición Profesional/análisis , Citocinas , Monitoreo del Ambiente/métodos , Factor de Necrosis Tumoral alfa , Estudios Transversales , Suero/química , Minería , Oro
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