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1.
Photodiagnosis Photodyn Ther ; 49: 104302, 2024 Aug 10.
Artículo en Inglés | MEDLINE | ID: mdl-39134252

RESUMEN

PURPOSE: The purpose of this study was to evaluate the correlations and consistency among the central corneal thickness (CCT) of healthy myopic patients measured with three different anterior segment analysis systems. DESIGN: This was a retrospective study. The study included myopia patients who had undergone preoperative examinations in the refractive surgery department of our hospital between January 2021 and December 2023. The CCT was measured separately using Pentacam, Sirius, and IOLMaster 700. METHODS: Statistical analysis was conducted using SPSS software. Correlations among the three groups of measured values were assessed using the Pearson method, and a simple scatter plot and fitting line were drawn. Bland‒Altman scatter plots and 95 % limits of agreement (LoAs) were used to evaluate consistency in the data among the systems. RESULTS: A total of 269 patients participated in the study, including 134 males (49.8 %) and 135 females (50.2 %). The CCT measurements by Pentacam, Sirius, and IOLMaster 700 instruments were found to be 541.63 ± 31.67 µm, 541.74 ± 33.36 µm, and 548.90 ± 34.19 µm respectively; significant differences were observed among these measurements (p < 0.05). Significant differences were also observed in CCT between Pentacam and IOLMaster 700 as well as between Sirius and IOLMaster 700 (p < 0.05). The CCT measurements by all three devices showed high positive correlation with all p values less than 0.001: Pentacam and Sirius, r = 0.972; Pentacam and IOLMaster 700, r = 0.966; and Sirius and IOLMaster 700, r = 0.962. The respective 95 % LoAs were -0.18∼0.18; -1.51∼-1.11; and -1.52∼-1. CONCLUSION: The results indicate that there is a high correlation in measuring CCT for healthy myopic eyes using three different anterior segment analysis systems. However, the differences in the values measured by the three devices were statistically significant. Therefore, in actual clinical practice, it is suggested that the same device should be used to measure and evaluate the CCT across visits.

2.
Sci Rep ; 14(1): 18308, 2024 08 07.
Artículo en Inglés | MEDLINE | ID: mdl-39112681

RESUMEN

Late gadolinium enhancement (LGE) is a widely used magnetic resonance imaging method for assessing cardiac disease. However, the relationship between different LGE signal thresholds and microscopic tissue staining images is unclear. In this study, we performed cardiovascular MRI on myocardial infarction (MI) model rats and evaluated the relationship between LGE with different signal thresholding methods and tissue staining images. We prepared 16 rats that underwent MRI 14-18 days following a surgery to create an MI model. We captured cine and LGE images of the cardiac short-axis and longitudinal two- and four-chamber views. The mean ± 2SD, ± 3SD, and ± 5SD of the pixel values in the non-infarcted area were defined as the LGE area. We compared areas of Sirius red staining, determined by the color tone, with their respective LGE areas at end-diastole and end-systole. We observed that the LGE area calculated as the mean ± 2SD of the non-infarcted area at end-diastole demonstrated a significant positive correlation with the area of Sirius red staining (Pearson's correlation coefficient in both: 0.81 [p < 0.01]). Therefore, the LGE area calculated as the mean ± 2SD of the non-infarcted area at end-diastole best reflected the MI area in tissue staining.


Asunto(s)
Medios de Contraste , Modelos Animales de Enfermedad , Gadolinio , Imagen por Resonancia Magnética , Infarto del Miocardio , Animales , Infarto del Miocardio/diagnóstico por imagen , Infarto del Miocardio/patología , Ratas , Imagen por Resonancia Magnética/métodos , Masculino , Coloración y Etiquetado/métodos , Miocardio/patología , Ratas Sprague-Dawley
3.
Eur J Surg Oncol ; 50(10): 108485, 2024 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-39047326

RESUMEN

BACKGROUND: Localization of non-palpable melanoma, Merkel cell carcinoma (MCC) and soft tissue sarcoma (STS) lesions can be difficult due to size, location, and obesity of patients or fibrosis due to previous treatments. Magnetic seed localization (MSL) is a common method to localize non-palpable breast lesions, but the feasibility of MSL for non-palpable melanoma, MCC and STS lesions has not yet been described. METHODS: In this retrospective single center cohort study, all consecutive patients between January 2021 and October 2023 who had a resection of a non-palpable melanoma, MCC or STS lesion guided by Sirius Pintuition, a MSL technique, were included. The primary endpoint was successful lesion localization during surgery and the secondary endpoints were seed migration, negative resection margins, and complications. RESULTS: Seventy-nine seeds were placed for 76 lesions, which were resected during 68 surgeries in 61 patients. All lesions (100 %) were localized and resected. Median time of surgery was 44 min. No seed migration was observed. A negative resection margin was achieved for 60 (78.9 %) lesions. Clavien Dindo grade ≥2 complications occurred in 7.4 %. CONCLUSION: Magnetic seed localization with Sirius Pintuition is feasible for both non-palpable melanoma, MCC, and STS lesions.

4.
Food Chem ; 455: 139769, 2024 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-38843716

RESUMEN

Bitter orange (Citrus aurantium) is an important source of essential oils with high antimicrobial activities, however the composition and antifungal potential of the decoction peels is little explored. This study assessed the peel decoction's chemical profile at the secondary metabolism level and its antifungal activity against the melon phytopathogen Fusarium jinanense. The decoction's antifungal potential was investigated using a bioassay-guided fractionation approach based on Solid-Phase Extraction (SPE) and LC-HRMS/MS analysis. Coumarins and flavones were the most abundant classes of compounds in the high-value fractions responsible for up to 61% of the mycelial inhibition of F. jinanense. Overall, this study has presented for the first time the chemical composition, the antifungal potential of the decoction of C. aurantium peels and the compounds associated with these results. This strategy can guide the exploration of under-explored food sources and add value to compounds or fractions enriched with bioactive compounds.


Asunto(s)
Citrus , Fusarium , Enfermedades de las Plantas , Extractos Vegetales , Fusarium/efectos de los fármacos , Enfermedades de las Plantas/microbiología , Extractos Vegetales/farmacología , Extractos Vegetales/química , Citrus/química , Citrus/microbiología , Cucurbitaceae/química , Cucurbitaceae/microbiología , Antifúngicos/farmacología , Antifúngicos/química , Espectrometría de Masas en Tándem , Extracción en Fase Sólida , Pruebas de Sensibilidad Microbiana
5.
Front Physiol ; 15: 1374309, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38860111

RESUMEN

Introduction: Isolation and confinement are significant stressors during space travel that can impact crewmembers' physical and mental health. Space travel has been shown to accelerate vascular aging and increase the risk of cardiovascular and cerebrovascular disorders. However, the effect of prolonged isolation and confinement on microvascular function has not yet been thoroughly investigated. Methods: Retinal vascular imaging was conducted on four crewmembers during- and post-8-month SIRIUS-21 space analog mission. Central retinal arteriolar equivalent (CRAE), central retinal venular equivalent (CRVE), and arteriovenous ratio (AVR) were measured. Pulse wave velocity (PWV), an indicator of arterial stiffness, was also measured. Results: Data from 4 participants was analyzed. These participants had a mean age of 34.75 ± 5.44 years, height of 170.00 ± 2.00 cm, weight of 74.50 ± 12.53 kg, and average BMI of 25.47 ± 3.94 kg/m2. During- and post-isolation, average CRVE showed an upward trend (Pearson's r 0.784, R-square 0.62), suggesting a dilation of retinal venules, while AVR showed a downward trend (Pearson's r -0.238, R-square 0.057), which is suggestive of a higher risk of cardiovascular and cerebrovascular dysfunctions. But neither of these trends were statistically significant. Additionally, the average PWV showed an upward trend during- and after-isolation across all crew members. Conclusion: Isolation and confinement appear to contribute towards retinal vascular damage and arterial stiffness. This cautiously suggests an increased risk of cardiovascular and cerebrovascular disorders due to the contribution of the isolation in space flight. Further studies are needed to confirm and expand on these results as we prepare for future manned missions to the Moon and Mars.

6.
Biol Methods Protoc ; 9(1): bpae027, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38800072

RESUMEN

Picrosirius red staining constitutes an important and broadly used tool to visualize collagen and fibrosis in various tissues. Although multiple qualitative and quantitative analysis methods to evaluate fibrosis are available, many require specialized devices and software or lack objectivity and scalability. Here, we aimed to develop a versatile and powerful "QuantSeg" macro in the FIJI image processing software capable of automated, robust, and quick collagen quantification in cardiac tissue from light micrographs. To examine different patterns of fibrosis, an optional segmentation algorithm was implemented. To ensure the method's validity, we quantified the collagen content in a set of wild-type versus plakoglobin-knockout murine hearts exhibiting extensive fibrosis using both the macro and an established, fluorescence microscopy-based method, and compared results. To demonstrate the capabilities of the segmentation feature, rat hearts were examined post-myocardial infarction. We found the QuantSeg macro to robustly detect the differences in fibrosis between knockout and control hearts. In sections with low collagen content, the macro yielded more consistent results than using the fluorescence microscopy-based technique. With its wide range of output parameters, ease of use, cost effectiveness, and objectivity, the QuantSeg macro has the potential to become an established method for analysis of PSR-stained tissue. The novel segmentation feature allows for automated evaluation of different patterns of cardiac fibrosis for the first time.

7.
Environ Sci Technol ; 58(22): 9828-9839, 2024 Jun 04.
Artículo en Inglés | MEDLINE | ID: mdl-38785362

RESUMEN

Pharmaceuticals and their human metabolites are contaminants of emerging concern in the aquatic environment. Most monitoring studies focus on a limited set of parent compounds and even fewer metabolites. However, more than 50% of the most consumed pharmaceuticals are excreted in higher amounts as metabolites than as parents, as confirmed by a literature analysis within this study. Hence, we applied a wide-scope suspect screening approach to identify human pharmaceutical metabolites in wastewater influent from three Swiss treatment plants. Based on consumption amounts and human metabolism data, a suspect list comprising 268 parent compounds and over 1500 metabolites was compiled. Online solid phase extraction combined with liquid chromatography coupled to high-resolution tandem mass spectrometry was used to analyze the samples. Data processing, annotation, and structure elucidation were achieved with various tools, including molecular networking as well as SIRIUS/CSI:FingerID and MetFrag for MS2 spectra rationalization. We confirmed 37 metabolites with reference standards and 16 by human liver S9 incubation experiments. More than 25 metabolites were detected for the first time in influent wastewater. Semiquantification with MS2Quant showed that metabolite to parent concentration ratios were generally lower compared to literature expectations, probably due to further metabolite transformation in the sewer system or limitations in the metabolite detection. Nonetheless, metabolites pose a large fraction to the total pharmaceutical contribution in wastewater, highlighting the need for metabolite inclusion in chemical risk assessment.


Asunto(s)
Aguas Residuales , Contaminantes Químicos del Agua , Aguas Residuales/química , Humanos , Contaminantes Químicos del Agua/metabolismo , Espectrometría de Masas en Tándem , Preparaciones Farmacéuticas/metabolismo , Cromatografía Liquida , Monitoreo del Ambiente/métodos , Extracción en Fase Sólida
8.
J Synchrotron Radiat ; 31(Pt 3): 547-556, 2024 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-38630437

RESUMEN

Fourth-generation synchrotron storage rings represent a significant milestone in synchrotron technology, offering outstandingly bright and tightly focused X-ray beams for a wide range of scientific applications. However, due to their inherently tight magnetic lattices, these storage rings have posed critical challenges for accessing lower-energy radiation, such as infrared (IR) and THz. Here the first-ever IR beamline to be installed and to operate at a fourth-generation synchrotron storage ring is introduced. This work encompasses several notable advancements, including a thorough examination of the new IR source at Sirius, a detailed description of the radiation extraction scheme, and the successful validation of our optical concept through both measurements and simulations. This optimal optical setup has enabled us to achieve an exceptionally wide frequency range for our nanospectroscopy experiments. Through the utilization of synchrotron IR nanospectroscopy on biological and hard matter samples, the practicality and effectiveness of this beamline has been successfully demonstrated. The advantages of fourth-generation synchrotron IR sources, which can now operate with unparalleled stability as a result of the stringent requirements for producing low-emittance X-rays, are emphasized.

9.
Methods Mol Biol ; 2788: 97-136, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38656511

RESUMEN

Plant specialized metabolites have diversified vastly over the course of plant evolution, and they are considered key players in complex interactions between plants and their environment. The chemical diversity of these metabolites has been widely explored and utilized in agriculture and crop enhancement, the food industry, and drug development, among other areas. However, the immensity of the plant metabolome can make its exploration challenging. Here we describe a protocol for exploring plant specialized metabolites that combines high-resolution mass spectrometry and computational metabolomics strategies, including molecular networking, identification of structural motifs, as well as prediction of chemical structures and metabolite classes.


Asunto(s)
Espectrometría de Masas , Metaboloma , Metabolómica , Plantas , Metabolómica/métodos , Plantas/metabolismo , Espectrometría de Masas/métodos , Biología Computacional/métodos
10.
Genome Biol ; 25(1): 77, 2024 03 22.
Artículo en Inglés | MEDLINE | ID: mdl-38519987

RESUMEN

BACKGROUND: B-type lamins are critical nuclear envelope proteins that interact with the three-dimensional genomic architecture. However, identifying the direct roles of B-lamins on dynamic genome organization has been challenging as their joint depletion severely impacts cell viability. To overcome this, we engineered mammalian cells to rapidly and completely degrade endogenous B-type lamins using Auxin-inducible degron technology. RESULTS: Using live-cell Dual Partial Wave Spectroscopic (Dual-PWS) microscopy, Stochastic Optical Reconstruction Microscopy (STORM), in situ Hi-C, CRISPR-Sirius, and fluorescence in situ hybridization (FISH), we demonstrate that lamin B1 and lamin B2 are critical structural components of the nuclear periphery that create a repressive compartment for peripheral-associated genes. Lamin B1 and lamin B2 depletion minimally alters higher-order chromatin folding but disrupts cell morphology, significantly increases chromatin mobility, redistributes both constitutive and facultative heterochromatin, and induces differential gene expression both within and near lamin-associated domain (LAD) boundaries. Critically, we demonstrate that chromatin territories expand as upregulated genes within LADs radially shift inwards. Our results indicate that the mechanism of action of B-type lamins comes from their role in constraining chromatin motion and spatial positioning of gene-specific loci, heterochromatin, and chromatin domains. CONCLUSIONS: Our findings suggest that, while B-type lamin degradation does not significantly change genome topology, it has major implications for three-dimensional chromatin conformation at the single-cell level both at the lamina-associated periphery and the non-LAD-associated nuclear interior with concomitant genome-wide transcriptional changes. This raises intriguing questions about the individual and overlapping roles of lamin B1 and lamin B2 in cellular function and disease.


Asunto(s)
Cromatina , Lamina Tipo B , Animales , Lamina Tipo B/genética , Heterocromatina , Hibridación Fluorescente in Situ , Lamina Tipo A/genética , Lamina Tipo A/metabolismo , Laminas , Expresión Génica , Mamíferos/genética
11.
3 Biotech ; 14(3): 93, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38433848

RESUMEN

Factors, namely pH, laccase-like activity, dyes concentration as well as 1-Hydroxybenzotriazole (HBT) concentration was examined. The results indicated that the maximum decolorization yield and rate reached 98.30 ± 0.10% and 5.84 ± 0.01%/min, respectively for Sirius Blue, and 99.34 ± 0.47% and 5.85 ± 0.12%/min, respectively for Sirius Red after 4 h. The presence of the redox mediator 1-hydroxybenzotriazole (HBT) greatly improved the decolorization levels. The optimum concentrations of HBT, dyes, and laccase were 0.62 mM, 50 mg/L, and 0.89 U/mL respectively at pH 4.58 for both dyes. Phytotoxicity tests using treated and untreated dyes proved that the applied treatment slightly decreased the toxicity of the by-products. However, the germination index (GI) increased from 14.6 to 36.08% and from 31.6 to 36.96% for Sirius Red and Sirius Blue, respectively. The present study focused on the treatment of two recalcitrant azo dyes, namely: Sirius Blue (Direct Blue 71) and Sirius Red (Direct Red 80). The decolorization was performed using cell-free supernatant from Coriolopsis gallica culture with high laccase activity. Response surface methodology (RSM) and Box-Behnken design were applied to optimize the decolorization of the two tested dyes. The effect of four.

12.
BMC Ophthalmol ; 24(1): 138, 2024 Mar 27.
Artículo en Inglés | MEDLINE | ID: mdl-38539111

RESUMEN

PURPOSE: To assess the level of agreement and evaluate the reliability of measurements between two Scheimpflug imaging modalities, Scansys (MediWorks, China) and Sirius (CSO, Italy), in quantifying the anterior segment parameters in healthy eyes. METHODS: In a cross-sectional study, the right eyes of 38 healthy participants without any ocular or systemic diseases were examined. A range of anterior segment parameters including anterior and posterior flat and steep keratometry, central corneal thickness (CCT), thinnest corneal thickness (TCT), anterior chamber depth (ACD), anterior chamber angle (ACA), corneal volume, anterior chamber volume, and horizontal white to white diameter, derived from the sagittal curvature maps were measured. To evaluate the reliability of the measurements, intraclass correlation coefficient (ICC) and correlation coefficient were measured. Additionally, Bland-Altman plots were employed to examine the agreement in mean (bias line) and 95% limits of agreement between the two devices. RESULTS: The mean age was 31.5 ± 6.9 (range: 19-47) years. The ICC indicated that the majority of anterior segment parameters had an excellent or good level of reliability, surpassing the threshold of 0.9. Nevertheless, CCT and ACA exhibited a moderate level of reliability, with ICC values of 0.794 and 0.728, respectively. The correlation analysis showed a strong correlation for all the variables tested. The Bland-Altman plots revealed that the bias line was near zero and the 95% limits of agreement were narrow for most variables, except for the anterior flat and steep keratometry, which were found to range from - 0.57 to 0.84 D and - 0.68 to 0.87 D, respectively. CONCLUSION: Scansys and Sirius devices can be effectively used interchangeably for the evaluation of most anterior segment parameters; however, for anterior corneal curvatures, CCT and ACA, their alternative use is not recommended.


Asunto(s)
Córnea , Tomografía , Humanos , Adulto Joven , Adulto , Reproducibilidad de los Resultados , Estudios Transversales , Estudios Prospectivos , Córnea/diagnóstico por imagen , Topografía de la Córnea/métodos , Tomografía de Coherencia Óptica
13.
Int Ophthalmol ; 44(1): 103, 2024 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-38376696

RESUMEN

PURPOSE: This study aimed to evaluate the consistency of preoperative keratometric values, anterior segment, and intraocular lens (IOL) power measurements in patients with cataract and no comorbidities using the Sirius topography device (CSO, Italy) and Lenstar LS 900 (Haag-Streit AG, Köeniz, Switzerland). METHODS: Patients with grade 2 and 3 cataracts who applied to Ophthalmology Clinic of Diskapi Yildirim Beyazit Education and Research Hospital, University of Health Sciences and planned for cataract surgery were included the study. Forty eyes with cataract from 40 patients were taken in the study. All patients underwent preoperative assessment using a combined Scheimpflug-Placido disc-based tomography device (Sirius) and Lenstar before cataract surgery. Keratometric measurements, such as flat keratometry (K1), steep keratometry (K2), and maximum keratometry (Kmax), and anterior segment parameters, white-to-white (WTW) distance, IOL power, astigmatism (AST), anterior chamber depth (ACD), aqueous depth (AD), and central cornea thickness (CCT), were recorded. RESULTS: There were significant differences between K1Lenstar and K1Sirius, K2Lenstar and K2Sirius, KmaxLenstar and KmaxSirius, WTWLenstar and WTWSirius, and IOL powerLenstar versus IOL powerSirius. However, there were insignificant differences between ASTLenstar and ASTSirius, ACDLenstar versus ACDSirius, ADLenstar and ADSirius, and CCTLenstar and CCTSirius variables. Furthermore, it was found that Sirius measured significantly higher than Lenstar, especially in terms of IOL power. CONCLUSION: Significant differences were observed between Lenstar and Sirius in terms of keratometric values, WTW distance, and IOL power. The IOL power value measured with Sirius was found to be higher than the IOL power value measured with Lenstar.


Asunto(s)
Astigmatismo , Extracción de Catarata , Catarata , Humanos , Biometría , Catarata/diagnóstico , Córnea
14.
Molecules ; 29(2)2024 Jan 18.
Artículo en Inglés | MEDLINE | ID: mdl-38257390

RESUMEN

The textile industry produces high volumes of colored effluents that require multiple treatments to remove non-adsorbed dyes, which could be recalcitrant due to their complex chemical structure. Most of the studies have dealt with the biodegradation of mono or diazo dyes but rarely with poly-azo dyes. Therefore, the aim of this paper was to study the biodegradation of a four azo-bond dye (Sirius grey) and to optimize its decolorization conditions. Laccase-containing cell-free supernatant from the culture of a newly isolated fungal strain, Coriolopsis gallica strain BS9 was used in the presence of 1-hydroxybenzotriazol (HBT) to optimize the dye decolorization conditions. A Box-Benken design with four factors, namely pH, enzyme concentration, HBT concentration, and dye concentration, was performed to determine optimal conditions for the decolorization of Sirius grey. The optimal conditions were pH 5, 1 U/mL of laccase, 1 mM of HBT, and 50 mg/L of initial dye concentration, ensuring a decolorization yield and rate of 87.56% and 2.95%/min, respectively. The decolorized dye solution showed a decrease in its phytotoxicity (Germination index GI = 80%) compared to the non-treated solution (GI = 29%). This study suggests that the laccase-mediator system could be a promising alternative for dye removal from textile wastewater.


Asunto(s)
Compuestos Azo , Lacasa , Polyporaceae , Compuestos Azo/toxicidad , Biodegradación Ambiental , Colorantes/toxicidad , Poli A
15.
Eur J Ophthalmol ; 34(1): 59-70, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-37731321

RESUMEN

PURPOSE: To determine the sensitive indices distinguishing forme-fruste keratoconus (FFKC) and early keratoconus (early KC) from thin normal corneas, and their cutoff values using Sirius topography. METHODS: 156 eyes with normal thin corneas < 500 um (group 1), 99 eyes with early KC (group 2) and 41 eyes with FFKC (group 3), were assessed retrospectively for: corneal keratometric indices, pachymetry indices, corneal aberrations, elevation indices; thinnest corneal point elevation, Q value, root mean square (RMS) withRMS/ area (RMS/A); and KC summary indices of front and back surfaces; surface asymmetry index (SIf, SIb), elevation at KC vertex (KVf, KVb), Baiocchi-Calossi-Versaci index (BCVf, BCVb) and summation of its vector (BCV). Cutoff values were calculated. RESULTS: Keratometry indices were significantly different between early KC and thin normal cases (apex front curvature had the highest area-under-the-ROC-curve (AUROC) (0.926) in early KC, while only apex curvature and coordinates were significant in FFKC cases. Pachymetry indices did not show any significance in differentiating either early KC or FFKC from normal thin corneas. KC summary indices were highly significant among the 3 groups. The highest AUROC was observed with KVb in early KC (0.987) and with KVf in FFKC (0.831). Vertical coma and vertical trefoil showed the highest significance of all aberration parameters differentiating the 3 groups. Thinnest point elevation, RMS and RMS/A showed the highest AUROC in differentiating early KC and FFKC cases from thin normal corneas. CONCLUSION: Comparing early KC and FFKC to thin normal corneas, Sirius provided high precision in prediction.


Asunto(s)
Queratocono , Humanos , Queratocono/diagnóstico , Topografía de la Córnea , Estudios Retrospectivos , Córnea , Curva ROC , Paquimetría Corneal
16.
Int J Mol Sci ; 24(24)2023 Dec 13.
Artículo en Inglés | MEDLINE | ID: mdl-38139263

RESUMEN

Tissue fibrosis is characterized by chronic fibroblast activation and consequently excessive accumulation of collagen-rich extracellular matrix. In vitro microplate-based assays are essential to investigate the underlying mechanism and the effect of antifibrotic drugs. In this study, in the absence of a gold-standard method, we optimized a simple, cost-effective, Sirius Red-based colorimetric measurement to determine the collagen production of fibroblasts grown on 96-well tissue culture plates. Based on our findings, the use of a serum-free medium is recommended to avoid aspecific signals, while ascorbate supplementation increases the collagen production of fibroblasts. The cell-associated collagens can be quantified by Sirius Red staining in acidic conditions followed by alkaline elution. Immature collagens can be precipitated from the culture medium by acidic Sirius Red solution, and after subsequent centrifugation and washing steps, their amount can be also measured. Increased attention has been paid to optimizing the assay procedure, including incubation time, temperature, and solution concentrations. The resulting assay shows high linearity and sensitivity and could serve as a useful tool in fibrosis-related basic research as well as in preclinical drug screening.


Asunto(s)
Colágeno , Colorantes , Humanos , Colorantes/farmacología , Colágeno/farmacología , Coloración y Etiquetado , Matriz Extracelular , Fibrosis , Fibroblastos
17.
Heliyon ; 9(11): e21841, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-38027816

RESUMEN

Ethnopharmacological relevance: Canarium schweinfurthii, also called ''Elemierd'Afrique'', is used in Cameroonian folk medicine (bark decoction) to treat patients suffering from hypertension.Aim of the study: This study aimed at evaluating the antihypertensive activities of the stem bark of Canarium schweinfurthii and identifying potential compounds present in its extract that may support or oppose its ethnomedicinial use. Materials and methods: Stem bark extract of Canarium schweinfurthii was prepared by maceration using 70 % ethanol followed by redissolution in methanol and hyphenated. Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS) analysis for the detection and characterisation of secondary metabolites. Antihypertensive effects were assessed in Wistar rats after induction of hypertension with sodium chloride (NaCl) 18 % at a dose of 0.01mL/gbody weight once a day for four weeks.Hemodynamic parameters were measured weekly by anon-invasive method using the CODA system. Results: The ethanolic bark extract of C. schweinfurthii significantly inhibited the increase of blood pressure with a maximum of 23.18 % (systolic pressure, p < 0.0001), 24.77 % (diastolic pressure, p < 0.001) and 22.95 % (mean pressure, p < 0.0001) at a dose of 200 mg/kgbody weight at the 4th week, compared to agroup of Wistar rats that received only NaCl (negative control). Similarly, the extract significantly inhibited the increase in heart rate by 18.84 % (p < 0.001) at 200 mg/kgbody weight at week four. Hematological parameters did not differ significantly between the extract-treated and control groups. The UPLC-MS/MS spectrometric analysis provided evidence for the presence of several C30 terpenoids containing three or five oxygen atoms and exhibiting pentacyclic triterpenoid structures, as well as C29 terpenoids and related compounds containing nitrogen in addition to oxygen, using spectral matching, and in silico molecular formula and structure prediction. Additionally, two features were annotated with high-confidence as lignans, structurally closely related to hinokinin and dehydrocubebin through MS/MS-based in silico structure prediction using CSI: Finger ID in SIRIUS5. The lignans have been previously reported from stem bark of plants belonging to the Burseraceae family. Conclusion: The ethanolic stem bark extract of C. schweinfurthii demonstrated antihypertensive properties on the tested Wistar rats. These results support the ethnopharmacological use of C. schweinfurthii concoctions for the treatment of hypertension and suggest a protective effect against salt damage, hypothetically by the up regulation of antioxidative enzymes and/or lipids, mitigatings membrane peroxidation.

18.
Eur J Clin Microbiol Infect Dis ; 42(11): 1355-1363, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37794128

RESUMEN

PURPOSE: This study aims to evaluate the performance of two latest generation matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) systems in routine laboratory settings, focusing on turnaround time (TAT), time to results (TTR), hands-on time, and identification rate. METHODS: We conducted a time and motion study on three workflow scenarios to simulate different laboratory settings. Overall, 618 bacterial isolates from a tertiary hospital's laboratory were processed using the VITEK MS PRIME (bioMérieux) and the MALDI Biotyper sirius (Bruker Daltonics) and their corresponding databases VITEK IVD Database 3.2 and MBT reference library 12. RESULTS: The target preparation process showed no significant difference in TAT, but the Biotyper workflow had a shorter hands-on time by 3 to 6 min. In the measurement process, TTR was three to five times shorter for the Biotyper sirius while hands-on time was significantly shorter for VITEK MS PRIME (approximately 1.5 min per target). The identification rate without retesting was 97.9% for VITEK MS PRIME and 98.9% for Biotyper sirius. Both systems achieved 100% agreement at genus and 96.2% at species level. CONCLUSION: Both systems exhibited excellent identification rates for routine bacterial isolates. Due to its high speed, the Biotyper sirius is suited for laboratories with high sample throughput and a workflow designed for processing larger batches. The VITEK MS PRIME, with its "load and go" system accommodating up to 16 targets, reduces hands-on time, making it a reasonable choice for laboratories with fewer identifications overall but a higher number of targets and a workflow designed for parallel processing on different workstations.


Asunto(s)
Laboratorios , Humanos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos
19.
Ther Adv Ophthalmol ; 15: 25158414231180984, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37434779

RESUMEN

Background: Cycloplegic drops are commonly used in ophthalmology practice. Changes in anterior segment parameters may occur after cycloplegia. These changes can be evaluated with corneal topography. Objective: This study aimed to compare the effects of 1% cyclopentolate hydrochloride and 1% tropicamide on anterior segment parameters using the Sirius Scheimpflug imaging technique. Design: A cross-sectional study. Methods: One hundred twenty eyes of sixty healthy volunteers with spherical equivalent (SE) values of 0 to ±1 diopter (D) were studied. The right eye of each subject had instillation of cyclopentolate hydrochloride 1% (Group 1) and the left eye of each subject had instillation of tropicamide 1% (Group 2). SE, intraocular pressure, and corneal topography measurements were performed before and 40 min after instillation were compared. Results: In Group 1, SE, aqueous depth, anterior chamber depth, iridocorneal angle (ICA), anterior chamber volume (ACV), and pupil size (PS) values were significantly increased (p < 0.001, p = 0.01, p < 0.001, p = 0.03, p < 0.001, and p < 0.001, respectively). In Group 2, SE, ICA, ACV, and PS were significantly increased (p < 0.001 for all). Keratometric values (K1 and K2) and central corneal thickness changed insignificantly in both groups (p > 0.05). The effects of the two administered agents on all parameters were similar (p > 0.05). Conclusions: Cyclopentolate hydrochloride and tropicamide affected SE, ICA, ACV, and PS values significantly. These parameters are important in intraocular lens (IOL) power calculations. PS is also important in refractive surgery and cataract surgery with multifocal IOL implantation. Although there was an insignificant difference between the agents, the effects of tropicamide on the parameters were smaller than those of cyclopentolate.

20.
Hepatol Int ; 17(5): 1162-1169, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37358741

RESUMEN

BACKGROUND AND AIMS: Liver fibrosis predicts adverse clinical outcomes, such as liver-related death (LRD) and hepatocellular carcinoma (HCC) in patients with non-alcoholic fatty liver disease (NAFLD). We aimed to investigate the accuracy of semi-automated quantification of collagen proportionate area (CPA) as an objective new method for predicting clinical outcomes. METHOD: Liver biopsies from patients with NAFLD underwent computerized image morphometry of Sirius Red staining with CPA quantification performed by ImageScope. Clinical outcomes, including total mortality, LRD, and combined liver outcomes (liver decompensation, HCC, or LRD), were determined by medical records and population-based data-linkage. The accuracy of CPA for predicting outcomes was compared with non-invasive fibrosis tests (Hepascore, FIB-4, APRI). RESULTS: A total of 295 patients (mean age 50 years) were followed for a median (range) of 9 (0.2-25) years totalling 3253 person-years. Patients with CPA ≥ 10% had significantly higher risks for total death [hazard ratio (HR): 5.0 (1.9-13.2)], LRD [19.0 (2.0-182.0)], and combined liver outcomes [15.6 (3.1-78.6)]. CPA and pathologist fibrosis staging (FS) showed similar accuracy (AUROC) for the prediction of total death (0.68 vs. 0.70), LRD (0.72 vs. 0.77) and combined liver outcomes (0.75 vs. 0.78). Non-invasive serum markers Hepascore, APRI, and FIB-4 reached higher AUROC; however, they were not statistically significant compared to that of CPA except for Hepascore in predicting total mortality (0.86 vs. 0.68, p = 0.009). CONCLUSION: Liver fibrosis quantified by CPA analysis was significantly associated with clinical outcomes including total mortality, LRD, and HCC. CPA achieved similar accuracy in predicting outcomes compared to pathologist fibrosis staging and non-invasive serum markers.


Asunto(s)
Carcinoma Hepatocelular , Neoplasias Hepáticas , Enfermedad del Hígado Graso no Alcohólico , Humanos , Persona de Mediana Edad , Enfermedad del Hígado Graso no Alcohólico/complicaciones , Carcinoma Hepatocelular/diagnóstico por imagen , Carcinoma Hepatocelular/complicaciones , Neoplasias Hepáticas/diagnóstico por imagen , Neoplasias Hepáticas/complicaciones , Cirrosis Hepática , Hígado/diagnóstico por imagen , Hígado/patología , Biomarcadores , Fibrosis , Biopsia , Índice de Severidad de la Enfermedad
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