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1.
J Mol Recognit ; 36(12): e3062, 2023 12.
Artículo en Inglés | MEDLINE | ID: mdl-37849017

RESUMEN

Glycation of biomolecules results in the formation of advanced glycation end products (AGEs). Immunoglobulin G (IgG) has been implicated in the progression of various diseases, including diabetes and cancer. This study purified three IgG subclasses (IgG1, IgG2, and IgG3) from Camelus dromedarius colostrum using ammonium sulfate fractionation and chromatographic procedures. SDS-PAGE was performed to confirm the purity and molecular weight of the IgG subclasses. Several biochemical and biophysical techniques were employed to study the effect of glycation on camel IgG using methylglyoxal (MGO), a dicarbonyl sugar. Early glycation measurement showed an increase in the fructosamine content by ~four-fold in IgG2, ~two-fold in IgG3, and a slight rise in IgG1. AGEs were observed in all classes of IgGs with maximum hyperchromicity (96.6%) in IgG2. Furthermore, glycation-induced oxidation of IgGs led to an increase in carbonyl content and loss of -SH groups. Among subclass, IgG2 showed the highest (39.7%) increase in carbonyl content accompanied by 82.5% decrease in -SH groups. Far UV-CD analysis illustrated perturbation of ß-sheet structure during glycation reaction with MGO. Moreover, glycation of IgG proceeds to various conformational states like aggregation and increased hydrophobicity. In addition, the cytotoxicity assay (MTT) illustrated the proliferation of breast cancer cells (MCF-7) with IgG2 treatment.


Asunto(s)
Camelus , Neoplasias , Animales , Reacción de Maillard , Óxido de Magnesio , Inmunoglobulina G/química , Productos Finales de Glicación Avanzada , Proliferación Celular
2.
Int J Mol Sci ; 23(20)2022 Oct 20.
Artículo en Inglés | MEDLINE | ID: mdl-36293474

RESUMEN

Nephrotic syndrome (NS) is a kidney illness characterized by excessive proteinuria, hypoalbuminemia, edema, and hyperlipidemia, which may lead to kidney failure and necessitate renal transplantation. End-stage renal disease, cardiovascular issues, and mortality are much more common in those with NS. Therefore, the present study aimed to identify potential new biomarkers associated with the pathogenesis and diagnosis of NS. The liquid chromatography-mass spectrometry (LC-MS) metabolomics approach was applied to profile the metabolome of human serum of patients with NS. A total of 176 metabolites were significantly altered in NS compared to the control. Arginine, proline, and tryptophan metabolism; arginine, phenylalanine, tyrosine, and tryptophan biosynthesis were the most common metabolic pathways dysregulated in NS. Furthermore, alanyl-lysine and isoleucyl-threonine had the highest discrimination between NS and healthy groups. The candidate biomarkers may lead to understanding the possible metabolic alterations associated with NS and serve as potential diagnostic biomarkers.


Asunto(s)
Síndrome Nefrótico , Humanos , Síndrome Nefrótico/diagnóstico , Lisina , Triptófano , Metabolómica/métodos , Metaboloma , Biomarcadores , Arginina , Tirosina , Prolina , Fenilalanina , Treonina
3.
Molecules ; 27(12)2022 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-35744792

RESUMEN

Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder characterized by progressive muscle loss, leading to difficulties in movement. Mutations in the DMD gene that code for the protein dystrophin are responsible for the development of DMD disorder, where the synthesis of this protein is completely halted. Therefore, circulating dystrophin protein could be a promising biomarker of DMD disease. Current methods for diagnosing DMD have sensitivity, specificity, and reproducibility limitations. Herein, a quantitative liquid chromatography-tandem spectrometry (LC-MS/MS) technique in multiple reaction monitoring (MRM) mode was designed and validated for accurate dystrophin protein measurement in a dried blood spot (DBS). The method was successfully validated on the basis of international guidelines regarding calibration curves, precision, and accuracy. In addition, patients and healthy controls were used to test the amount of dystrophin protein circulating in DBS samples as a potential biomarker for DMD disorders. DMD patients were found to have considerably lower levels than controls. To the best of our knowledge, this is the first study to report dystrophin levels in DBS through LC-MS/MS as a diagnostic marker for DMD to the proposed MRM method, providing a highly specific and sensitive approach to dystrophin quantification in a DBS that can be applied in DMD screening.


Asunto(s)
Distrofina , Distrofia Muscular de Duchenne , Biomarcadores/metabolismo , Cromatografía Liquida , Distrofina/genética , Humanos , Distrofia Muscular de Duchenne/genética , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem
4.
Inflammopharmacology ; 30(3): 737-748, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35364736

RESUMEN

Rheumatoid arthritis (RA) is a chronic multifactorial disease, provocative, and degenerative autoimmune condition that impacts millions of individuals around the globe. As a result of this understanding, anti-inflammatory drugs have been created, perhaps widely effective (like steroids) and highly specialized methods (including anti-TNF antibody) using biological therapies (including TNF inhibitors). Despite this, the connections between inflammatory response, articular development, and intracellular responsiveness to changes in oxygen concentration are undervalued in rheumatoid arthritis. Hypoxia, or a lack of oxygen, is thought to cause enhanced synovial angiogenesis in RA, which is mediated by some of the hypoxia-inducible factors like vascular endothelial growth factor (VEGF). Substantial genetic alterations occur when the HIF regulatory factors signaling cycle is activated, allowing organelles, tissues, and species to acclimatize to decreasing oxygen saturation. The most well-characterized hypoxia-responsive transcripts are the angiogenic stimulant VEGF, whose production is greatly elevated by hypoxia in several types of cells, especially RA synovium fibroblasts. Blocking vascular endothelial growth factors has been demonstrated to be helpful in murine models of rheumatism, indicating how hypoxia could trigger the angiogenesis process, resulting in the progression of RA. These mechanisms highlight the intimate affiliation amongst hypoxia, angiogenesis, and inflammation in rheumatoid arthritis. This review will look at how hypoxia activates molecular pathways and how other pathways involving inflammatory signals develop and sustain synovitis in rheumatoid arthritis.


Asunto(s)
Artritis Reumatoide , Factor A de Crecimiento Endotelial Vascular , Animales , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/metabolismo , Humanos , Hipoxia/metabolismo , Ratones , Neovascularización Patológica/metabolismo , Oxígeno/uso terapéutico , Inhibidores del Factor de Necrosis Tumoral , Factor A de Crecimiento Endotelial Vascular/metabolismo
5.
Medicina (Kaunas) ; 58(3)2022 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-35334523

RESUMEN

Background and Objectives: In this study, we aimed to investigate the link between common -2548G>A (rs7799039) promoter variant of the human leptin gene (LEP) with leptin and serum glucose leptin levels in obese Saudi patients. Materials and Methods: A total of 206 Saudi adults (80 obese normotensive nondiabetics, 76 obese hypertensive with Type 2 Diabetes and 50 normotensive nondiabetic controls) were genotyped for -2548G>A LEP polymorphism using the polymerase chain reaction-restriction fragment-length polymorphism technique. Results: Participants with minor AA genotype had significantly higher blood glucose levels (6.8 ± 0.55 mmol/L vs. 5.8 ± 0.30 mmol/L; p < 0.04) and HOMA-IR (4.1 ± 0.84 vs. 2.6 ± 0.67; p = 0.03) against those carrying major GG genotype. Participants with heterozygous GA genotype had significantly higher serum leptin levels against those carrying major GG genotype (40.0 ± 2.6 ng/mL vs. 29.6 ± 2.6 ng/mL; p = 0.04). Further investigation showed that individuals with AA, GA, GA + AA genotypes are at greater risk of developing hyperglycemia compared to those with GG genotype [OR 3.7(1.6−8.4), p = 0.001; 3.2 (1.2−8.6), p = 0.03; 3.5 (1.6−7.7), p = 0.001, respectively]. Additionally, the -2548AA allele was shown to be a risk factor for hyperglycemia [OR 1.9 (1.2−3.0), p = 0.006]. Our data revealed no relationship between this variant of the LEP gene with systolic and diastolic BP, signifying that this genetic variant is not a significant marker of obesity and hypertension in the Saudi population. Conclusions: AA and GA genotypes and LEP gene -2548AA alleles may signify potent risk factors predisposing healthy individuals to develop T2DM regardless of blood-pressure profile.


Asunto(s)
Diabetes Mellitus Tipo 2 , Leptina/genética , Adulto , Presión Sanguínea , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/genética , Frecuencia de los Genes , Glucosa , Humanos , Obesidad/complicaciones , Obesidad/genética , Polimorfismo de Nucleótido Simple , Arabia Saudita/epidemiología
6.
Medicina (Kaunas) ; 58(8)2022 Jul 25.
Artículo en Inglés | MEDLINE | ID: mdl-35893101

RESUMEN

Background and Objectives: Folic acid (FA) is a necessary ingredient for numerous bodily activities including pregnancy. Because of this, women should have knowledge and awareness of the health benefits of FA supplementation. Thus, we aimed to investigate the level of knowledge on the importance of FA and determine associated factors for knowledge among female college students at King Saud University in Riyadh, Saudi Arabia. Material and Methods: We conducted a cross-sectional study using a questionnaire between January 2020 and February 2021 among female college students aged 17 to 26 years old. The questionnaire adapted with permission from Alnaami et al. included questions on the demographic profile of the participants as well as questions related to their knowledge and awareness of FA, FA supplementation, the importance of supplementation and their sources of knowledge of FA. Results: A total of 437 female undergraduate students participated in the study, 285 (65.2%) of whom were from the non-health colleges and 152 (34.8%) from the health colleges. The majority of participants were between ages 17 and 21 years old (n = 361, 82.6%). Half of the respondents were in their 3rd and 4th year of study (n = 122, 50.8%), 138 respondents (31.6%) were married, and 111 of these married women (80.4%) had children. There were 266 respondents (61.0%) who had heard and had knowledge of FA, 241 (55.3%) knew of FA timing of intake, 243 (55.7%) of FA duration of intake and 362 (83.0%) knew of the diseases prevented by FA supplementation. Linear regression analysis showed that being in the health college (B = 1.464, t = 11.37, p < 0.001, 95% CI = 1.211, 1.717) and a higher educational year level (B = 0.139, t = 2.442, p = 0.015, 95% CI = 0.027, 0.251) were the significant predictors of knowledge of FA. Conclusions: Knowledge of FA and FA supplementation was low at 61% considering that our study population were college students. Being enrolled in a health college and in a higher educational year level were significant positive correlates of higher knowledge of FA and FA supplementation. Despite this, there exists a gap of information regarding FA and FA supplementation particularly among single women and college educated women in the early years of their college life as well as those in non-health colleges.


Asunto(s)
Ácido Fólico , Conocimientos, Actitudes y Práctica en Salud , Adolescente , Adulto , Niño , Estudios Transversales , Suplementos Dietéticos , Femenino , Ácido Fólico/uso terapéutico , Humanos , Embarazo , Estudiantes , Adulto Joven
7.
J Proteome Res ; 20(3): 1558-1570, 2021 03 05.
Artículo en Inglés | MEDLINE | ID: mdl-33557525

RESUMEN

Dexamethasone is a synthetic glucocorticoid medication vastly used to treat abnormal immune responses and inflammation. Although the medication is well-established in the medical community, the prolonged treatment with high dosages of dexamethasone may lead to severe adverse effects through mechanisms that are not yet well-known. Lipids are a large class of hydrophobic molecules involved in energy storage, signaling, modulation of gene expression, and membranes. Hence, untargeted lipidomics may help unravel the biochemical alterations following prolonged treatment with high dosages of dexamethasone. We performed comprehensive lipidomic analyses of brain, heart, kidney, liver, and muscle samples obtained from rats that were treated with intramuscular injections of dexamethasone for 14 weeks compared to healthy controls. The employed methodology and statistical analysis showed that phosphatidic acids, glycerophospholipids, plasmalogens, and fatty acids are deeply affected by prolonged use of the medication. Brain tissue was only mildly affected, but skeletal muscle showed a strong accumulation of lipids that may be correlated with alterations in the energy metabolism, myopathy, and oxidative processes. This work provides new insights into the mechanisms of action and adverse effects for one of the most commonly prescribed class of drugs in the world.


Asunto(s)
Lipidómica , Lípidos , Animales , Dexametasona/efectos adversos , Ácidos Grasos , Glicerofosfolípidos , Ratas
9.
Cureus ; 16(4): e59364, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38817460

RESUMEN

Objectives This cross-sectional analytical study aims to evaluate medical students' awareness and satisfaction regarding the utilization of simulation-based learning (SBL) as a method for clinical teaching at King Saud University (KSU) over the past 12 months. It seeks to understand how such learning methods enhance students' self-satisfaction and clinical skills, facilitate the application of learned knowledge, and assess the role of instructors in providing ample practice opportunities in the skills laboratory. Furthermore, the study aims to assess the satisfaction levels of students in both preclinical and clinical years regarding the time allocated for skills laboratory sessions and the integration of high-fidelity technology in simulation-based training programs at KSU. Methods In this cross-sectional study, a total of 306 male and female medical students from the College of Medicine at KSU participated, comprising 196 preclinical students (first, second, and third years) and 110 clinical students (fourth and fifth years). Quantitative data was collected through a structured questionnaire on a 5-point Likert scale that showed degrees of satisfaction. The satisfaction was measured based on a 5-point Likert scale that shows the degree of satisfaction from (very dissatisfied, dissatisfied, neither dissatisfied nor satisfied, satisfied, and very satisfied), and we calculated the p-value based on an independent t-test, and the percentage represented the percentage of students who chose satisfied and very satisfied. Results The results showed overall satisfaction with SBL (mean: 3.98, 71.10%), and it was recognized as a useful and effective method of learning skills. It is reported that it helped the students implement what they learned. At the same time, lower satisfaction was identified in areas with less allocated time for skill labs. Moreover, lack of accessibility and lack of trained staff were reported, and they should be addressed by providing staff with proper training. Conclusion The results of the study will help to understand how students' learning needs should be addressed. Moreover, providing simulation-based training is a pathway compliant with the best educational standards that should be adapted according to each institution's singularities. Besides offering further results, the study presents suggestions for further research.

10.
Clin Chim Acta ; 557: 117861, 2024 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-38490341

RESUMEN

BACKGROUND: Glutaric aciduria type-1 (GA-1) is a rare metabolic disorder due to glutaryl coenzyme A dehydrogenase deficiency, causing elevated levels of glutaryl-CoA and its derivatives. GA-1 exhibits symptoms like macrocephaly, developmental delays, and movement disorders. Timely diagnosis through genetic testing and newborn screening is crucial. However, in some cases, transiently elevated level of glutarylcarnitine (C5DC) challenges accurate diagnosis, highlighting the need for alternative diagnostic methods, like mass spectrometry-based untargeted metabolomics, to identify additional biomarkers for distinguishing falsely suspected GA-1 from healthy newborns. METHODOLOGY: DBS samples from falsely suspected GA-1 newborns (n = 47) and matched control were collected through the NBS program. Untargeted metabolomics using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) was performed to enable biomarker and pathway investigations for significantly altered metabolites. RESULTS: 582 and 546 were up- and down-regulated metabolites in transient GA-1. 155 endogenous metabolites displayed significant variations compared to the control group. Furthermore, our data identified novel altered metabolic biomarkers, such as N-palmitoylcysteine, heptacarboxyporphyrin, 3-hydroxylinoleoylcarnitine, and monoacylglyceride (MG) (0:0/20:1/0:0), along with perturbed metabolic pathways like sphingolipid and thiamine metabolism associated with the transient elevated C5DC levels in DBS samples. CONCLUSIONS: A distinct metabolic pattern linked to the transient C5DC elevation in newborns was reported to enhance the prediction of the falsely positive cases, which could help avoiding unnecessary medical treatments and minimizing the financial burdens in the health sector.


Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos , Encefalopatías Metabólicas , Glutaril-CoA Deshidrogenasa/deficiencia , Humanos , Recién Nacido , Glutaril-CoA Deshidrogenasa/genética , Encefalopatías Metabólicas/diagnóstico , Encefalopatías Metabólicas/genética , Encefalopatías Metabólicas/terapia , Biomarcadores , Errores Innatos del Metabolismo de los Aminoácidos/diagnóstico , Errores Innatos del Metabolismo de los Aminoácidos/genética , Metabolómica
11.
Adv Respir Med ; 92(3): 241-253, 2024 Jun 04.
Artículo en Inglés | MEDLINE | ID: mdl-38921063

RESUMEN

BACKGROUND: Pneumonia is a ubiquitous health condition with severe outcomes. The advancement of ultrasonography techniques allows its application in evaluating pulmonary diseases, providing safer and accessible bedside therapeutic decisions compared to chest X-ray and chest computed tomography (CT) scan. Because of its aforementioned benefits, we aimed to confirm the diagnostic accuracy of lung ultrasound (LUS) for pneumonia in adults. METHODS: A systematic literature search was performed of Medline, Cochrane and Crossref, independently by two authors. The selection of studies proceeded based on specific inclusion and exclusion criteria without restrictions to particular study designs, language or publication dates and was followed by data extraction. The gold standard reference in the included studies was chest X-ray/CT scan or both. RESULTS: Twenty-nine (29) studies containing 6702 participants were included in our meta-analysis. Pooled sensitivity, specificity and PPV were 92% (95% CI: 91-93%), 94% (95% CI: 94 to 95%) and 93% (95% CI: 89 to 96%), respectively. Pooled positive and negative likelihood ratios were 16 (95% CI: 14 to 19) and 0.08 (95% CI: 0.07 to 0.09). The area under the ROC curve of LUS was 0. 9712. CONCLUSIONS: LUS has high diagnostic accuracy in adult pneumonia. Its contribution could form an optimistic clue in future updates considering this condition.


Asunto(s)
Neumonía , Ultrasonografía , Humanos , Neumonía/diagnóstico por imagen , Ultrasonografía/métodos , Adulto , Sensibilidad y Especificidad , Pulmón/diagnóstico por imagen
12.
Artículo en Inglés | MEDLINE | ID: mdl-37668758

RESUMEN

Oxidative stress caused due to the perturbations in the oxidant-antioxidant system can damage molecules and cause cellular alteration leading to the pathogenesis of multiple diseases. This study was designed and performed to investigate the antioxidant and anti-inflammatory effects of an alkaloid, gramine on H2O2-induced oxidative stress on HEK 293 cells. Cell viability and morphometric analysis of cells treated with H2O2 and gramine were studied. Oxidative stress and inflammatory and antioxidant enzymes such as ROS, LPO, NO, SOD, GSH, and CAT were analyzed. Furthermore, mRNA expression of SOD, CAT, and COX-2 was also evaluated. H2O2 at concentration > 0.3 mM and gramine at concentration > 80 µg/mL affect the proliferation. Viability and morphometric analysis showed that gramine has protective effects. Treating cells with gramine suppressed oxidative stress and inflammatory enzymes, whereas antioxidant enzymes were enhanced. SOD and CAT mRNA levels were overexpressed and COX-2 mRNA levels were decreased in the treated groups. Gramine possesses effective antioxidant potential and can regulate oxidative stress and damages associated with it.

13.
Sci Rep ; 13(1): 6161, 2023 04 15.
Artículo en Inglés | MEDLINE | ID: mdl-37061630

RESUMEN

Chronic kidney disease (CKD) is a serious public health problem characterized by progressive kidney function loss leading to end-stage renal disease (ESRD) that demands dialysis or kidney transplantation. Early detection can prevent or delay progression to ESRD. The study aimed to gain new insights into the perturbed biochemical reactions and to identify novel distinct biomarkers between ESRD and CKD. Serum samples of 32 patients with ESRD (n = 13) and CKD (n = 19) were analyzed using chemical isotope labeling liquid chromatography-mass spectrometry metabolomics approach. A total of 193 metabolites were significantly altered in ESRD compared to CKD and were mainly involved in aminoacyl-tRNA biosynthesis, branched-chain amino acid (BCAA) biosynthesis, taurine metabolism, and tryptophan metabolism. Three kynurenine derivatives, namely, 2-aminobenzoic acid, xanthurenic acid, and hydroxypicolinic acid were upregulated in ESRD compared to CKD due to the significant decrease in glomerular filtration rate with the progression of CKD to ESRD. N-Hydroxy-isoleucine, 2-aminobenzoic acid, and picolinic acid yielded AUC > 0.99 when analyzed using Receiver Operating Characteristic (ROC) analysis. Our findings suggest that inhibiting the kynurenine pathway might be a promising target to delay CKD progression and that metabolites with high discriminative ability might serve as potential prognostic biomarkers to monitor the progression of CKD to ESRD or used in combination with current markers to indicate the status of kidney damage better.


Asunto(s)
Fallo Renal Crónico , Insuficiencia Renal Crónica , Humanos , Quinurenina , Diálisis Renal , Factores de Riesgo , Biomarcadores/análisis , Progresión de la Enfermedad , Tasa de Filtración Glomerular
14.
Metabolites ; 13(8)2023 Aug 11.
Artículo en Inglés | MEDLINE | ID: mdl-37623881

RESUMEN

According to studies, the microbiome may contribute to the emergence and spread of breast cancer. E. coli is one of the Enterobacteriaceae family recently found to be present as part of the breast tissue microbiota. In this study, we focused on the effect of E. coli secretome free of cells on MCF-7 metabolism. Liquid chromatography-mass spectrometry (LC-MS) metabolomics was used to study the E. coli secretome and its role in MCF-7 intra- and extracellular metabolites. A comparison was made between secretome-exposed cells and unexposed controls. Our analysis revealed significant alterations in 31 intracellular and 55 extracellular metabolites following secretome exposure. Several metabolic pathways, including lactate, aminoacyl-tRNA biosynthesis, purine metabolism, and energy metabolism, were found to be dysregulated upon E. coli secretome exposure. E. coli can alter the breast cancer cells' metabolism through its secretome which disrupts key metabolic pathways of MCF-7 cells. These microbial metabolites from the secretome hold promise as biomarkers of drug resistance or innovative approaches for cancer treatment, either as standalone therapies or in combination with other medicines.

15.
Turk J Gastroenterol ; 34(8): 859-865, 2023 08.
Artículo en Inglés | MEDLINE | ID: mdl-37485558

RESUMEN

BACKGROUND/AIMS: Irritable bowel syndrome is prevalent in the general population. This study investigates the association between dietary intake and irritable bowel syndrome in medical college students at King Saud University besides its prevalence. MATERIALS AND METHODS: This is an analytical cross-sectional study of 426 students (271 males and 155 females, age 21.21 ± 1.58 years) from 5 academic levels of King Saud University Medical College. A self-reported questionnaire for Rome IV criteria was completed by each participant. They also filled out a food frequency questionnaire to assess their nutritional intake. RESULTS: The overall prevalence of irritable bowel syndrome was 17.8% without correlation to age and academic year in Medical School. However, the prevalence was higher in females than in males (40/115 vs. 36/235, P = .001). The irritable bowel syndrome group consumed significantly more energy, carbohydrates, and saturated fatty acids, while the non-irritable bowel syndrome group consumed significantly more fibers and niacin (P < .001 and P = .005, respectively). CONCLUSION: About 17.8% of medical students had irritable bowel syndrome with a greater prevalence in females. The irritable bowel syndrome group consumed significantly more energy, carbohydrates, and saturated fatty acids, while the non-irritable bowel syndrome group consumed significantly more fibers and niacin. Our results did not show any significant association between irritable bowel syndrome and fermentable oligosaccharide, disaccharide, monosaccharide, and polyol intake. Overall, both groups were not adhering to the Saudi dietary recommended intake.


Asunto(s)
Síndrome del Colon Irritable , Niacina , Estudiantes de Medicina , Masculino , Femenino , Humanos , Adulto Joven , Adulto , Estudios Transversales , Ciudad de Roma , Síndrome del Colon Irritable/epidemiología , Encuestas y Cuestionarios , Carbohidratos , Prevalencia
16.
J Pharm Biomed Anal ; 216: 114801, 2022 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-35504217

RESUMEN

The cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel found on the apical surface of epithelial cells in the airway and gastrointestinal tract. A mutation in the CFTR protein is responsible for developing cystic fibrosis (CF) disease. Therefore, circulating CFTR protein could be a promising biomarker of CF disease. Multiple methodological challenges are associated with CF's available diagnostic and screening methods, such as low specificity and potential false discovery rate, mainly for ethnic groups whose CFTR mutations are not covered in the mutation panels. Herein, we have developed an absolute quantification (AQUA) method based on two CFTR signature peptides (SPs). A liquid chromatography-tandem spectrometry (LC-MS/MS) method in multiple reaction monitoring (MRM) mode (MRM transitions 1168.90 > 85.929 and 707.19 > 85.93 of SP1 and SP2, respectively) enabled the accurate quantification of CFTR protein in a dried blood spot (DBS). The method was validated successfully based on international guidelines in terms of signal linearity, precision (within-run CV 3.37-8.54%; between-run CV 5.15-11.06% for the selected SPs), and accuracy (within-run 93.4-105.59%; between-run 97.45-103.28% for the selected SPs). The level of soluble CFTR protein was evaluated as a potential biomarker for CF using patients (n = 39) and healthy controls (n = 30), were found to be in CF patients lower than controls. For instant, the level of signature peptide 1 (SP1) was 2.09 ± 0.55 nM, 68.77 ± 1.40 nM in CF patients compared to Ctrl, respectively; p < 0.0001. This study is the first to report CFTR levels in DBS using signature peptides by LC-MS/MS as a diagnostic marker for CF. The receiver operating characteristic (ROC) for CFTR SP1 and SP2 showed a significant area under the curves (AUC) 0.7714 (99% CI, p < 0.0001), and 0.8234 (99% CI, p < 0.0001), respectively. The presented MRM method provides a highly specific and sensitive approach to CFTR quantification in a DBS and could be applied in CF screening.


Asunto(s)
Regulador de Conductancia de Transmembrana de Fibrosis Quística , Fibrosis Quística , Biomarcadores , Cromatografía Liquida , Fibrosis Quística/diagnóstico , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Regulador de Conductancia de Transmembrana de Fibrosis Quística/metabolismo , Humanos , Espectrometría de Masas en Tándem
17.
Diagnostics (Basel) ; 12(2)2022 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-35204412

RESUMEN

Different laboratory methods are used to measure serum ferritin levels as a marker of iron status in the general population. This study aimed to compare serum ferritin levels using enzyme-linked immunosorbent assay (ELISA) versus immunochemiluminescence (Cobas e411) and immunoturbidimetric (Cobas Integra 400) methods in terms of sensitivity, specificity and accuracy, and whether they can be used interchangeably. A comparative cross-sectional study enrolled one hundred and six adult Yemeni patients (33 males and 73 females) aged 18-55 years, recruited from the dermatology and cosmetic center of Hadhramout Modern Hospital, Mukalla, Yemen. Serum ferritin levels were measured using ELISA, Cobas e411, and Cobas Integra 400 methods. For method comparison, a paired-sample t-test was used. For the consistency between the three methods, they were analyzed with regression and Pearson correlation coefficient. For determining accuracy, a receiver operating curve (ROC) was used. Bias error between the methods was determined through a Bland-Altman plot analysis. Our results did not show any significant statistical difference between ELISA and Cobas e411 (52.55 ± 7.4 µg/L vs. 52.58 ± 7.5 µg/L, p = 0.967), while there were significantly higher values from Cobas Integra 400 results than Cobas e411 (56.31 ± 7.8 µg/L vs. 52.58 ± 7.5 µg/L, p < 0.001) and ELISA (52.55 ± 7.4 µg/L vs. 56.31 ± 7.8 µg/L, p < 0.001). According to the correlation coefficient and linear regression analysis, a strong association between ELISA with Cobas e411 (r = 0.993, p < 0.001) and Cobas Integra 400 results (r = 0.994, p < 0.001) were revealed. For determining accuracy, Cobas e411 and Cobas Integra 400 results showed higher sensitivity (92.0%; 90.0%) and specificity (97.7%; 99.9%) respectively. Additionally, the Bland-Altman plot analysis showed a high agreement between the ELISA and Cobas e411 methods (bias: -0.035). In contrast, there was a low agreement between the ELISA and Cobas Integra 400 methods (bias: -3.75). Similarly, the agreement between Cobas e411 and Cobas Integra 400 methods was low (bias: -3.72). Serum ferritin levels were measured by Cobas e411, and Cobas Integra 400 methods were strongly correlated with the ELISA results, with higher sensitivity, specificity, and accuracy. However, further investigations with larger samples are required for improved accuracy and more precise results, and to determine whether they can be used interchangeably.

18.
Infect Dis Rep ; 14(6): 841-854, 2022 Nov 14.
Artículo en Inglés | MEDLINE | ID: mdl-36412744

RESUMEN

Numerous measures have been taken to slow the Coronavirus disease (COVID-19) rapid spread. Such population control techniques may have a substantial impact on people's attitudes, knowledge, and perception of COVID-19. This web-based cross-sectional survey aimed to assess Knowledge, Attitude, and Practices (KAP) towards COVID-19 among Hadhramout University Medical Students in Yemen from 15 June to 26 June 2020. This survey was performed using social media via the Google Platform among 422 Hadhramout University Medical students. After consenting, participants completed an online survey assessing sociodemographic data, 21 knowledge items, 15 attitudes items, and 5 perception items towards COVID-19. Of the total 422 participants, 389 (92.18%) were surveyed online, and 256 (65.8%) were females, and 133 (34.2%) were males aged 19-24 years (88.7%), studying medicine (58.9%), and living in urban areas (80.7%). The survey revealed that 64.0% of participants had good knowledge about the disease and 52.7% had positive attitudes towards protective measures against the virus. The majority of participants (98.2%) thought that the virus was transmitted through nasal droplets, and 59.6% agreed that the disease is dangerous. The majority of participants agreed that fever (99.2%), dry cough (97.9%), and difficulty breathing (99.5%) are the most common symptoms of the disease. The survey also showed high knowledge levels about preventive measures against the virus spreading, such as regular proper hand hygiene (99.7%), maintaining an appropriate distance (99.2%), avoiding touching eyes and nose (98.7%), and wearing facemasks in public places (97.4%). Moreover, 69.7% of participants agreed to be isolated at home if they got an infected person, 64.3% implemented washing hands with soap and water, 41.9% agreed to be separated at the hospital until they proved free from the disease, 46.0% agreed to inform the health authorities if they had any symptoms associated with the disease. By using sample T-test and analysis of variance (ANOVA), mean knowledge score about COVID-19 was significantly higher in males than in females (p = 0.029). Additionally, medicine students had significantly higher mean knowledge score than students of medical laboratory (p < 0.001) and nursing (p = 0.008). In general, our research revealed that participants had favorable opinions regarding the disease's preventative measures and a good awareness of it. However, more educational initiatives and campaigns that take into account KAP modifying elements are needed.

19.
Comput Biol Med ; 138: 104893, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34598069

RESUMEN

Understanding the underlying molecular mechanism of transporter activity is one of the major discussions in structural biology. A transporter can exclusively transport one ion (specific transporter) or multiple ions (general transporter). This study compared categorical and numerical features of general and specific calcium transporters using machine learning and attribute weighting models. To this end, 444 protein features, such as the frequency of dipeptides, organism, and subcellular location, were extracted for general (n = 103) and specific calcium transporters (n = 238). Aliphatic index, subcellular location, organism, Ile-Leu frequency, Glycine frequency, hydrophobic frequency, and specific dipeptides such as Ile-Leu, Phe-Val, and Tyr-Gln were the key features in differentiating general from specific calcium transporters. Calcium transporters in the cell outer membranes were specific, while the inner ones were general; additionally, when the hydrophobic frequency or Aliphatic index is increased, the calcium transporter act as a general transporter. Random Forest with accuracy criterion showed the highest accuracy (88.88% ±5.75%) and high AUC (0.964 ± 0.020), based on 5-fold cross-validation. Decision Tree with accuracy criterion was able to predict the specificity of calcium transporter irrespective of the organism and subcellular location. This study demonstrates the precise classification of transporter function based on sequence-derived physicochemical features.


Asunto(s)
Aprendizaje Automático
20.
J Family Med Prim Care ; 10(12): 4493-4496, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-35280641

RESUMEN

Objectives: To study the knowledge, attitude and practice level among young Saudi adults regarding osteoporosis. Methods: A cross-sectional study was done in Riyadh among Saudis, within the age group (17-30 years old), with sample size (663). A validated questionnaire was distributed in paper and electronic forms. The questionnaire had five parts: first part was the demographic variables, second part the source of the participant knowledge about the disease, third part assessed the knowledge about osteoporosis, fourth part assessed osteoporosis attitudes, and the fifth part assessed the practice for prevention of osteoporosis. The collected data was analysed by Statistical Package for the Social Sciences program. Results: It was found that there was low level of knowledge among overall participants; there was no significant difference between male and female in level of knowledge, while the age group 21-25 had the highest knowledge level among all other age groups. Males have better level of practice of a healthy lifestyle that decreases the risk of osteoporosis than females. In total, 52.5% of males have a sufficient level of practice, while only 34.9% of females have acceptable level of practice. Conclusion: The study found no significant difference between males and females regarding the knowledge of osteoporosis. Males were found to have better level of practice of a healthy lifestyle that decreases the level of osteoporosis then females. The study recommends similar studies and undertake more efforts in promotion of preventive programs for young adults.

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