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1.
J Biomed Opt ; 29(Suppl 2): S22702, 2025 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38434231

RESUMEN

Significance: Advancements in label-free microscopy could provide real-time, non-invasive imaging with unique sources of contrast and automated standardized analysis to characterize heterogeneous and dynamic biological processes. These tools would overcome challenges with widely used methods that are destructive (e.g., histology, flow cytometry) or lack cellular resolution (e.g., plate-based assays, whole animal bioluminescence imaging). Aim: This perspective aims to (1) justify the need for label-free microscopy to track heterogeneous cellular functions over time and space within unperturbed systems and (2) recommend improvements regarding instrumentation, image analysis, and image interpretation to address these needs. Approach: Three key research areas (cancer research, autoimmune disease, and tissue and cell engineering) are considered to support the need for label-free microscopy to characterize heterogeneity and dynamics within biological systems. Based on the strengths (e.g., multiple sources of molecular contrast, non-invasive monitoring) and weaknesses (e.g., imaging depth, image interpretation) of several label-free microscopy modalities, improvements for future imaging systems are recommended. Conclusion: Improvements in instrumentation including strategies that increase resolution and imaging speed, standardization and centralization of image analysis tools, and robust data validation and interpretation will expand the applications of label-free microscopy to study heterogeneous and dynamic biological systems.


Asunto(s)
Técnicas Histológicas , Microscopía , Animales , Citometría de Flujo , Procesamiento de Imagen Asistido por Computador
2.
Oral Health Prev Dent ; 22: 237-248, 2024 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-38989777

RESUMEN

PURPOSE: To assess adherence to follow-up maintenance visits among patients who had previously undergone crown-lengthening surgery and investigate the different factors impacting their compliance. MATERIALS AND METHODS: A total of 314 patients were identified for follow-up appointments. Based on their responses, participants were categorised into four groups: attendees, non-attendees, refusals, and unreachable. Furthermore, data on sociodemographic factors (age, sex, nationality, marital status, education, occupation, and residential area), medical history, dental history (including missing teeth, implants, or orthodontic treatment history), and past appointment attendance (average yearly appointments, missed appointment percentage, and last appointment date) were collected and analysed to understand their influence on patient compliance. RESULTS: In a sample of 314 patients, 102 (32.5%) attended the appointments successfully. Improved attendance rates were significantly associated with being female, Saudi Arabian, married, and employed (p < 0.05). Moreover, patients with a high frequency of annual appointments and a recent history of appointments exhibited better compliance. None of the analysed dental factors affected the attendance rates. CONCLUSION: About one-third of patients who had undergone crown lengthening surgery were compliant with the follow-up visits. Different factors influenced this compliance pattern to varying extents, with more efforts needed to enhance patients' commitment to these visits.


Asunto(s)
Alargamiento de Corona , Estado Civil , Cooperación del Paciente , Humanos , Femenino , Masculino , Adulto , Persona de Mediana Edad , Factores Sexuales , Empleo/estadística & datos numéricos , Adulto Joven , Factores de Edad , Escolaridad , Estudios de Seguimiento , Citas y Horarios , Ocupaciones , Adolescente
3.
Ann Afr Med ; 23(3): 365-371, 2024 Jul 01.
Artículo en Francés, Inglés | MEDLINE | ID: mdl-39034560

RESUMEN

BACKGROUND: Pressure ulcers (PUs) are a common global problem. They affect health-care costs and patient health because of increased morbidity and mortality rates. Critically ill patients are more prone to acquiring PUs than other patients. As the prevalence of PUs is high in Saudi Arabia, more attention is required for PU prevention. Nurses' knowledge and attitudes toward PU prevention are crucial. AIMS: This study aimed to assess the knowledge and attitudes of nursing students regarding PU prevention evidence-based guidelines. SETTING AND DESIGN: This descriptive, cross-sectional study was conducted among nursing students at Jazan University. MATERIALS AND METHODS: Data were collected using an electronic questionnaire. STATISTICAL ANALYSIS USED: Data were entered, coded, and analyzed using the SPSS version 23. RESULTS: A total of 378 participants were included. The mean age was 22.4 ± 3.4 years, and the mean knowledge score was 7.6 ± 2.64 (range = 0-16). Of the participants, 365 (96.6%) and 13 (3.4%) were considered to have poor and good knowledge about PUs, respectively. Only 72 (19%) correctly identified a lack of oxygen as the cause of PUs; other participants answered malnutrition and moisture. Conversely, 200 (52.9%) participants indicated that recent weight loss below patients' ideal weight increases the risk for PUs. Concerning knowledge about preventive measures, 64 (16.9%) participants correctly answered that a backward sitting position, with both legs resting on a footrest, can reduce the amount of pressure. Meanwhile, 169 (44.7%) participants correctly selected thick air cushion as a means to reduce the magnitude of pressure when patients are sliding down on a chair. Some participants answered that PUs can be reduced if patients are mobilized. Seventy-two (19%) participants indicated that patients laying on a visco-elastic foam mattress should be repositioned every 2 h to reduce the duration of pressure. Age, year of education, training experience, and department were not significantly associated with knowledge about PUs (P = 0.333, P = 0.370, P = 0.700, and P = 0.810, respectively). CONCLUSIONS: The general knowledge and attitudes toward PUs of the nursing students at Jazan University were poor. Nevertheless, they had an average knowledge level regarding PU prevention. Age and training experience were not significantly associated with knowledge about PUs.


Résumé Contexte:Les ulcères de pression (UP) sont un problème mondial courant. Ils affectent les coûts de santé et la santé des patients en raison de l'augmentation des taux de morbidité et de mortalité. Les patients gravement malades sont plus susceptibles de développer des UP que d'autres patients. Comme la prévalence des UP est élevée en Arabie saoudite, une attention accrue est nécessaire pour la prévention des UP. Les connaissances et attitudes des infirmiers en matière de prévention des UP sont cruciales.Objectifs:Cette étude visait à évaluer les connaissances et attitudes des étudiants en soins infirmiers concernant les directives de prévention des UP basées sur des preuves. Cadre et conception : Cette étude descriptive et transversale a été menée auprès des étudiants en soins infirmiers de l'Université de Jazan.Matériels et méthodes:Les données ont été collectées à l'aide d'un questionnaire électronique. Analyse statistique utilisée: Les données ont été saisies, codées et analysées à l'aide de la version 23 du logiciel SPSS.Résultats:Au total, 378 participants ont été inclus. L'âge moyen était de 22,4 ± 3,4 ans et le score moyen de connaissance était de 7,6 ± 2,64 (plage = 0-16). Parmi les participants, 365 (96,6 %) avaient une connaissance médiocre et 13 (3,4 %) une bonne connaissance des UP. Seuls 72 (19 %) ont correctement identifié un manque d'oxygène comme étant la cause des UP; les autres participants ont répondu la malnutrition et l'humidité. En revanche, 200 (52,9 %) participants ont indiqué qu'une perte de poids récente en dessous du poids idéal des patients augmente le risque de UP. En ce qui concerne les connaissances sur les mesures préventives, 64 (16,9 %) participants ont répondu correctement qu'une position assise inclinée vers l'arrière, avec les deux jambes reposant sur un repose-pieds, peut réduire la pression. Pendant ce temps, 169 (44,7 %) participants ont correctement sélectionné un coussin d'air épais comme moyen de réduire l'ampleur de la pression lorsque les patients glissent sur une chaise. Certains participants ont répondu que les UP peuvent être réduits si les patients sont mobilisés. Soixante-douze (19 %) participants ont indiqué que les patients allongés sur un matelas en mousse viscoélastique devraient être repositionnés toutes les 2 heures pour réduire la durée de la pression. L'âge, l'année d'éducation, l'expérience de formation et le département n'étaient pas significativement associés aux connaissances sur les UP (P = 0,333, P = 0,370, P = 0,700 et P = 0,810, respectivement).Conclusions:Les connaissances générales et les attitudes à l'égard des UP des étudiants en soins infirmiers de l'Université de Jazan étaient médiocres. Néanmoins, ils avaient un niveau de connaissance moyen concernant la prévention des UP. L'âge et l'expérience de formation n'étaient pas significativement associés aux connaissances sur les UP.


Asunto(s)
Conocimientos, Actitudes y Práctica en Salud , Úlcera por Presión , Estudiantes de Enfermería , Humanos , Úlcera por Presión/prevención & control , Úlcera por Presión/epidemiología , Estudiantes de Enfermería/psicología , Arabia Saudita , Estudios Transversales , Femenino , Masculino , Adulto , Encuestas y Cuestionarios , Adulto Joven , Universidades
4.
Retina ; 44(8): 1279-1287, 2024 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-39047122

RESUMEN

PURPOSE: To evaluate the reliability of clinical grading of diabetic retinopathy (DR) severity compared with grading on ultra-widefield pseudocolor fundus (UWF-CF) and ultra-widefield fluorescein angiography (UWF-FA) images and their relative detection of sight-threatening DR and referable DR. METHODS: A total of 184 diabetic eyes were analyzed. UWF-CF and UWF-FA images were graded based on the International Clinical Diabetic Retinopathy severity scale. Agreement between clinical and UWF-based severity grading was evaluated using Cohen's kappa coefficient. The rate of sight-threatening DR and referable DR was evaluated for each grading method. RESULTS: Moderate agreement was found between clinical grading and UWF-CF (k = 0.456, P < 0.001) and between UWF-CF and UWF-FA (k = 0.443, P < 0.001). The agreement between clinical grading and UWF-FA was fair (k = 0.397, P < 0.001). UWF-based grading identified a higher DR grade in 56 eyes (30%) on UWF-CF and 85 eyes (46.2%) on UWF-FA. Compared with clinical grading, UWF-FA detected a higher rate of sight-threatening DR (44%; 81/184 vs. 22.3%; 41/184), while UWF-CF detected more referable eyes (58.1%; 107/184 vs. 45.65%; 84/184). CONCLUSION: Ultra-widefield pseudocolor fundus is a valuable tool for identifying referable eyes and can be a useful, noninvasive adjunct to clinical grading. The results suggest that UWF-FA is particularly useful for detecting unsuspected sight-threatening DR in eyes with clinically referable DR.


Asunto(s)
Retinopatía Diabética , Angiografía con Fluoresceína , Índice de Severidad de la Enfermedad , Humanos , Retinopatía Diabética/diagnóstico , Retinopatía Diabética/clasificación , Reproducibilidad de los Resultados , Femenino , Masculino , Persona de Mediana Edad , Angiografía con Fluoresceína/métodos , Anciano , Estudios Retrospectivos , Adulto , Fondo de Ojo
5.
Cureus ; 16(5): e60866, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38910608

RESUMEN

Preeclampsia, a complex multisystem disorder predominantly impacting the kidneys and liver, manifests through hypertension and organ dysfunction in expectant mothers. Preeclampsia can also cause ocular signs, but they are uncommon. Exudative retinal detachment (ERD) is one such unusual but dangerous consequence. A thorough clinical description and therapy of a patient who experienced exudative retinal detachment while experiencing preeclampsia are provided in this study. A 28-year-old Saudi female, with no medical or surgical history, underwent an emergency cesarean section (CS) due to severe preeclampsia and failed induction of labor. The patient complained of painless blurry vision, with central dark spot and decreased vision starting from labor daytime. The patient was admitted to the hospital for blood pressure monitoring and further investigations. The patient was started on hydralazine intravenous (IV) and labetalol PO to control BP. The patient was delivered by cesarean section for preeclampsia with severe features after the failure of labor induction, and she had improved her vision by four weeks postpartum. Retinal detachment as a consequence of preeclampsia is conservatively managed, with a generally favorable prognosis. Previous studies have consistently emphasized the critical importance of a multidisciplinary approach that fosters collaboration between obstetricians and ophthalmologists. This collaborative strategy not only ensures comprehensive care but also facilitates early detection, timely intervention, and improved management outcomes for conditions affecting both maternal health and ophthalmic well-being during pregnancy.

6.
Cytometry A ; 2024 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-38943226

RESUMEN

Autofluorescence lifetime imaging microscopy (FLIM) is sensitive to metabolic changes in single cells based on changes in the protein-binding activities of the metabolic co-enzymes NAD(P)H. However, FLIM typically relies on time-correlated single-photon counting (TCSPC) detection electronics on laser-scanning microscopes, which are expensive, low-throughput, and require substantial post-processing time for cell segmentation and analysis. Here, we present a fluorescence lifetime-sensitive flow cytometer that offers the same TCSPC temporal resolution in a flow geometry, with low-cost single-photon excitation sources, a throughput of tens of cells per second, and real-time single-cell analysis. The system uses a 375 nm picosecond-pulsed diode laser operating at 50 MHz, alkali photomultiplier tubes, an FPGA-based time tagger, and can provide real-time phasor-based classification (i.e., gating) of flowing cells. A CMOS camera produces simultaneous brightfield images using far-red illumination. A second PMT provides two-color analysis. Cells are injected into the microfluidic channel using a syringe pump at 2-5 mm/s with nearly 5 ms integration time per cell, resulting in a light dose of 2.65 J/cm2 that is well below damage thresholds (25 J/cm2 at 375 nm). Our results show that cells remain viable after measurement, and the system is sensitive to autofluorescence lifetime changes in Jurkat T cells with metabolic perturbation (sodium cyanide), quiescent versus activated (CD3/CD28/CD2) primary human T cells, and quiescent versus activated primary adult mouse neural stem cells, consistent with prior studies using multiphoton FLIM. This TCSPC-based autofluorescence lifetime flow cytometer provides a valuable label-free method for real-time analysis of single-cell function and metabolism with higher throughput than laser-scanning microscopy systems.

7.
bioRxiv ; 2024 Jun 10.
Artículo en Inglés | MEDLINE | ID: mdl-38915614

RESUMEN

Autofluorescence microscopy uses intrinsic sources of molecular contrast to provide cellular-level information without extrinsic labels. However, traditional cell segmentation tools are often optimized for high signal-to-noise ratio (SNR) images, such as fluorescently labeled cells, and unsurprisingly perform poorly on low SNR autofluorescence images. Therefore, new cell segmentation tools are needed for autofluorescence microscopy. Cellpose is a deep learning network that is generalizable across diverse cell microscopy images and automatically segments single cells to improve throughput and reduce inter-human biases. This study aims to validate Cellpose for autofluorescence imaging, specifically from multiphoton intensity images of NAD(P)H. Manually segmented nuclear masks of NAD(P)H images were used to train new Cellpose models. These models were applied to PANC-1 cells treated with metabolic inhibitors and patient-derived cancer organoids (across 9 patients) treated with chemotherapies. These datasets include co-registered fluorescence lifetime imaging microscopy (FLIM) of NAD(P)H and FAD, so fluorescence decay parameters and the optical redox ratio (ORR) were compared between masks generated by the new Cellpose model and manual segmentation. The Dice score between repeated manually segmented masks was significantly lower than that of repeated Cellpose masks (p<0.0001) indicating greater reproducibility between Cellpose masks. There was also a high correlation (R2>0.9) between Cellpose and manually segmented masks for the ORR, mean NAD(P)H lifetime, and mean FAD lifetime across 2D and 3D cell culture treatment conditions. Masks generated from Cellpose and manual segmentation also maintain similar means, variances, and effect sizes between treatments for the ORR and FLIM parameters. Overall, Cellpose provides a fast, reliable, reproducible, and accurate method to segment single cells in autofluorescence microscopy images such that functional changes in cells are accurately captured in both 2D and 3D culture.

9.
Transl Vis Sci Technol ; 13(6): 8, 2024 Jun 03.
Artículo en Inglés | MEDLINE | ID: mdl-38874974

RESUMEN

Purpose: Both hypertension and diabetes are known to increase the wall-to-lumen ratio (WLR) of retinal arterioles, but the differential effects are unknown. Here, we study the timing and relative impact of hypertension versus diabetes on the WLR in diabetic retinopathy (DR) to address this unresolved question. Methods: This prospective cross-sectional study compared the retinal arteriolar WLR in 17 healthy eyes, 15 with diabetes but no apparent DR (DM no DR), and 8 with diabetic macular edema (DME) and either nonproliferative or proliferative DR. We imaged each arteriole using adaptive optics scanning laser ophthalmoscopy and measured the WLR using ImageJ. Multiple linear regression (MLR) was performed to estimate the effects of hypertension, diabetes, and age on the WLR. Results: Both subjects with DM no DR and subjects with DME had significantly higher WLR than healthy subjects (0.36 ± 0.08 and 0.42 ± 0.08 vs. 0.29 ± 0.07, 1-way ANOVA P = 0.0009). MLR in healthy subjects and subjects with DM no DR showed hypertension had the strongest effect (regression coefficient = 0.08, P = 0.009), whereas age and diabetes were not significantly correlated with WLR. MLR in all three groups together (healthy, DM no DR, and DME) showed diabetes had the strongest effect (regression coefficient = 0.05, P = 0.02), whereas age and hypertension were not significantly correlated with WLR. Conclusions: Hypertension may be an early driver of retinal arteriolar wall thickening in preclinical DR, independent of age or diabetes, whereas changes specific to DR may drive wall thickening in DME and later DR stages. Translational Relevance: We offer a framework for understanding the relative contributions of hypertension and diabetes on the vascular wall, and emphasize the importance of hypertension control early in diabetes even before DR onset.


Asunto(s)
Retinopatía Diabética , Hipertensión , Oftalmoscopía , Humanos , Estudios Transversales , Masculino , Retinopatía Diabética/patología , Femenino , Persona de Mediana Edad , Estudios Prospectivos , Arteriolas/patología , Arteriolas/diagnóstico por imagen , Hipertensión/complicaciones , Hipertensión/patología , Anciano , Adulto , Arteria Retiniana/patología , Arteria Retiniana/diagnóstico por imagen , Edema Macular/patología , Edema Macular/diagnóstico por imagen , Edema Macular/etiología
10.
Biomed Pharmacother ; 177: 117015, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38936196

RESUMEN

Injury of a peripheral nerve (PNI) leads to both ischemic and inflammatory alterations. Sciatic nerve injury (SNI) represents the most widely used model for PNI. Mesenchymal stem cell-based therapy (MSCs) has convenient properties on PNI by stimulating the nerve regeneration. Melatonin has cytoprotective activity. The neuroprotective characteristics of MSCs and melatonin separately or in combination remain a knowledge need. In the rats-challenged SNI, therapeutic roles of intralesional MSCs and intraperitoneal melatonin injections were evaluated by functional assessment of peripheral nerve regeneration by walking track analysis involving sciatic function index (SFI) and two electrophysiological tests, electromyography and nerve conduction velocity, as well as measurement of antioxidant markers in serum, total antioxidant capacity (TAC) and malondialdehyde, and mRNA expression of brain derived neurotrophic factor (BDNF) in nerve tissues in addition to the histopathological evaluation of nerve tissue. Both individual and combination therapy with MSCs and melatonin therapies could effectively ameliorate this SNI and promote its regeneration as evidenced by improving the SFI and two electrophysiological tests and remarkable elevation of TAC with decline in lipid peroxidation and upregulation of BDNF levels. All of these led to functional improvement of the damaged nerve tissues and good recovery of the histopathological sections of sciatic nerve tissues suggesting multifactorial synergistic approach of the concurrent usage of melatonin and MSCs in PNI. The combination regimen has the most synergistic neuro-beneficial effects in PNI that should be used as therapeutic option in patients with PNI to boost their quality of life.


Asunto(s)
Antioxidantes , Melatonina , Trasplante de Células Madre Mesenquimatosas , Regeneración Nerviosa , Traumatismos de los Nervios Periféricos , Nervio Ciático , Animales , Melatonina/farmacología , Traumatismos de los Nervios Periféricos/tratamiento farmacológico , Traumatismos de los Nervios Periféricos/terapia , Trasplante de Células Madre Mesenquimatosas/métodos , Ratas , Nervio Ciático/efectos de los fármacos , Nervio Ciático/lesiones , Regeneración Nerviosa/efectos de los fármacos , Masculino , Antioxidantes/farmacología , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/metabolismo , Modelos Animales de Enfermedad , Ratas Wistar , Conducción Nerviosa/efectos de los fármacos , Ratas Sprague-Dawley
11.
bioRxiv ; 2024 May 18.
Artículo en Inglés | MEDLINE | ID: mdl-38798331

RESUMEN

Autofluorescence lifetime imaging microscopy (FLIM) is sensitive to metabolic changes in single cells based on changes in the protein-binding activities of the metabolic co-enzymes NAD(P)H. However, FLIM typically relies on time-correlated single-photon counting (TCSPC) detection electronics on laser-scanning microscopes, which are expensive, low-throughput, and require substantial post-processing time for cell segmentation and analysis. Here, we present a fluorescence lifetime-sensitive flow cytometer that offers the same TCSPC temporal resolution in a flow geometry, with low-cost single-photon excitation sources, a throughput of tens of cells per second, and real-time single-cell analysis. The system uses a 375nm picosecond-pulsed diode laser operating at 50MHz, alkali photomultiplier tubes, an FPGA-based time tagger, and can provide real-time phasor-based classification ( i.e ., gating) of flowing cells. A CMOS camera produces simultaneous brightfield images using far-red illumination. A second PMT provides two-color analysis. Cells are injected into the microfluidic channel using a syringe pump at 2-5 mm/s with nearly 5ms integration time per cell, resulting in a light dose of 2.65 J/cm 2 that is well below damage thresholds (25 J/cm 2 at 375 nm). Our results show that cells remain viable after measurement, and the system is sensitive to autofluorescence lifetime changes in Jurkat T cells with metabolic perturbation (sodium cyanide), quiescent vs. activated (CD3/CD28/CD2) primary human T cells, and quiescent vs. activated primary adult mouse neural stem cells, consistent with prior studies using multiphoton FLIM. This TCSPC-based autofluorescence lifetime flow cytometer provides a valuable label-free method for real-time analysis of single-cell function and metabolism with higher throughput than laser-scanning microscopy systems.

12.
Nat Commun ; 15(1): 4097, 2024 May 16.
Artículo en Inglés | MEDLINE | ID: mdl-38755144

RESUMEN

Angiogenesis, the growth of new blood vessels from pre-existing vasculature, is essential for the development of new organ systems, but transcriptional control of angiogenesis remains incompletely understood. Here we show that FOXC1 is essential for retinal angiogenesis. Endothelial cell (EC)-specific loss of Foxc1 impairs retinal vascular growth and expression of Slc3a2 and Slc7a5, which encode the heterodimeric CD98 (LAT1/4F2hc) amino acid transporter and regulate the intracellular transport of essential amino acids and activation of the mammalian target of rapamycin (mTOR). EC-Foxc1 deficiency diminishes mTOR activity, while administration of the mTOR agonist MHY-1485 rescues perturbed retinal angiogenesis. EC-Foxc1 expression is required for retinal revascularization and resolution of neovascular tufts in a model of oxygen-induced retinopathy. Foxc1 is also indispensable for pericytes, a critical component of the blood-retina barrier during retinal angiogenesis. Our findings establish FOXC1 as a crucial regulator of retinal vessels and identify therapeutic targets for treating retinal vascular disease.


Asunto(s)
Barrera Hematorretinal , Células Endoteliales , Factores de Transcripción Forkhead , Neovascularización Retiniana , Animales , Masculino , Ratones , Angiogénesis , Barrera Hematorretinal/metabolismo , Células Endoteliales/metabolismo , Factores de Transcripción Forkhead/metabolismo , Factores de Transcripción Forkhead/genética , Cadena Pesada de la Proteína-1 Reguladora de Fusión/metabolismo , Cadena Pesada de la Proteína-1 Reguladora de Fusión/genética , Transportador de Aminoácidos Neutros Grandes 1/metabolismo , Transportador de Aminoácidos Neutros Grandes 1/genética , Ratones Endogámicos C57BL , Ratones Noqueados , Pericitos/metabolismo , Retina/metabolismo , Neovascularización Retiniana/metabolismo , Neovascularización Retiniana/genética , Neovascularización Retiniana/patología , Vasos Retinianos/metabolismo , Serina-Treonina Quinasas TOR/metabolismo
13.
Ophthalmol Sci ; 4(5): 100482, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38751454

RESUMEN

Purpose: To localize early capillary perfusion deficits in patients with diabetes mellitus (DM) without clinical diabetic retinopathy (DR) using averaged OCT angiography (OCTA). Design: Retrospective cross-sectional study. Participants: Patients with DM without DR and healthy controls. Methods: We measured perfusion deficits in the full retina, superficial capillary plexus (SCP), and deep capillary plexus (DCP) on averaged 3 × 3-mm OCTA images. Perfusion deficits were defined as the percentage of retinal tissue located >30 µm from blood vessels, excluding the foveal avascular zone (FAZ). One eye from each patient was selected based on image quality. We measured deficits in the parafoveal region, the 300 µm surrounding the FAZ, and 300 to 1000 µm surrounding the FAZ. If a capillary layer within one of these regions was significantly different in DM without DR compared with controls, we further characterized the location of perfusion deficit as periarteriolar, perivenular, or the capillaries between these 2 zones. Main Outcome Measures: Location of increased perfusion deficits in patients with DM without DR compared with controls. Results: Sixteen eyes from 16 healthy controls were compared with 16 eyes from 16 patients with DM without DR (age 45.1 ± 10.7 and 47.4 ± 15.2 years respectively, P = 0.64). Foveal avascular zone area and perfusion deficits in the entire parafovea and the 300 to 1000-µm ring around the FAZ were not significantly different between groups (P > 0.05 for all). Perfusion deficits in 300 µm around the FAZ were significantly increased in patients with DM without DR in full retinal thickness, SCP, and DCP (P < 0.05 for all). When analyzing the perivenular, periarteriolar, and capillary zones, only the perivenular DCP perfusion deficits were significantly increased (5.03 ± 2.92% in DM without DR and 2.73 ± 1.97% in controls, P = 0.014). Conclusions: Macular perfusion deficits in patients with DM without DR were significantly increased in the region nearest the FAZ, mainly at the perivenular deep capillaries. Further research on these early changes may improve our understanding of the capillaries most susceptible to vascular injury and disruption during diabetes. Financial Disclosures: Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

14.
Cureus ; 16(4): e58384, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38628380

RESUMEN

BACKGROUND AND OBJECTIVES: Stem cell banking (SCB) is a promising area of modern medicine with the potential to yield innovative treatments and cures. To effectively educate parents and implement laws and regulations that address parental concerns and encourage informed decision-making, it is imperative to emphasize parental viewpoints and their consequences for future healthcare. The study aims to establish the Saudi Arabian population's level of understanding regarding SCB and to comprehend the elements influencing parental knowledge, attitudes, and SCB decision-making processes. METHODOLOGY: A cross-sectional study was conducted among the population in the Makkah region of Saudi Arabia. Demographic data, knowledge levels, attitudes, and decision-making variables were gathered from 380 respondents. RESULTS: The study reveals a lack in their comprehension of the objectives and possible uses of SCB, together with the main sources of information on those banks and conveniently available banking choices. It showed varied results regarding attitudes about considering an SCB for their children. In addition, the majority of respondents had not made a consent decision about SCB for their children. It also illuminates the factors that could influence participants' decisions about SCB for their children and shows that a lack of information and understanding is the main obstacle faced by parents regarding SCB. It highlights that participants were generally in favor of learning more about SCB for their children. CONCLUSIONS: This study broadens our understanding of parental decision-making toward SCB and clarifies the elements influencing parents' opinions and worries and offers significant ramifications for lawmakers, medical professionals, and SCB. These implications can be utilized to enhance communication strategies, create instructional programs, and ease the fears of concerned parents.

15.
Open Vet J ; 14(1): 370-388, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38633191

RESUMEN

Background: Mastitis in goats is unquestionably a grave concern, with far-reaching implications for both animal well-being and productivity, while also presenting a potential threat to public health. Aim: The study aimed to compare culture methods and multiplex PCR (m-PCR) in the detection of the most three common mastitis-causing pathogens (Staphylococcus aureus, Escherichia coli, and Streptococcus spp.) and investigate the gene expression, single nucleotide polymorphisms (SNPs), serum concentrations of immunological and antioxidant indicators linked to mastitis in Shami goats. Methods: One hundred Shami do (50 Shami goats with clinical mastitis and 50 normal goats taken as control group). The culture methods and m-PCR analysis were used to find the bacteria in the milk samples. Blood samples were obtained to assess some hemato-biochemical parameters, detect SNPs, and determine the expression of certain immunological and antioxidant indicators in the genes. Results: The culture method detected the pathogens causing mastitis in 90% of the milk samples, but m-PCR detected them in 100% of the milk samples. SNPs linked to mastitis resistance/susceptibility in examined does were detected through DNA sequencing of immunological and antioxidant indicators. The magnitude of gene expression varied significantly between the resistant and mastitis-affected groups. Significant (P ˂ 0.05) elevations were noticed in WBCs count, mainly neutsrophils count, serum levels of BHB, NEFA, triglycerides, LDL-C, AST, ALT, ALP, creatinine, total protein, globulin, Ca, K, GPx, MDA, acute phase proteins, and cytokines in mastitis affected does as compared to control. While RBCs count, PCV%, lymphocytes count, serum concentration of glucose, cholesterol, HDL-C, albumin, Na, Cl, P, GSH, SOD, and catalase significantly (P ˂ 0.05) diminished in mastitis affected does compared to healthy ones. APPs and pro-inflammatory cytokines scored high sensitivities and NPVs but TNF-α and serum amyloid A (SAA) had the highest percentages of increase. Conclusion: The study confirmed that m-PCR is the most sensitive method for bacteria identification (S. aureus, E. coli, and Strept. spp.) while SNPs in antioxidant and immunological genes may be important genetic indicators for mastitis risk or resistance in Shami does. To establish an effective management plan and forecast the most sensitive risk time for illness onset, gene expression profiles of the tested genes may also be employed as proxy biomarkers. TNF-α and SAA may be precious indicators for the detection of caprine mastitis.


Asunto(s)
Enfermedades de las Cabras , Mastitis , Femenino , Animales , Antioxidantes , Cabras , Staphylococcus aureus , Factor de Necrosis Tumoral alfa , Egipto , Escherichia coli , Bacterias , Mastitis/microbiología , Mastitis/veterinaria , Genómica , Enfermedades de las Cabras/microbiología
16.
Invest Ophthalmol Vis Sci ; 65(4): 37, 2024 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-38652648

RESUMEN

Purpose: Adjuvant, pre-operative intravitreal anti-vascular endothelial growth factor (anti-VEGF) injections have been used to reduce peri-operative bleeding in eyes undergoing pars-plana vitrectomy for complications of proliferative diabetic retinopathy (PDR). To address the concern over their potential off-target effects of progressive fibrous contraction, we sought to dissect the transcriptional changes in the surgically extracted fibrovascular membranes (FVMs). Methods: We analyzed surgically extracted FVMs from 10 eyes: 4 eyes pretreated with intravitreal bevacizumab (IVB) and 6 untreated eyes. FVMs were digested into single cells, mRNA was extracted from endothelial cell-enriched (microbead selection with CD31) and non-endothelial cell compartments, followed by RT-qPCR quantification. We then compared the relative expression of genes involved in angiogenesis, endothelial cell integrity, and myofibroblastic processes between treated and untreated FVMs. Results: Endothelial cells from IVB pretreated FVMs showed significant reduction of VEGFA, VEGF receptors (FLT1 and KDR), and angiopoietin 2 expression as well as increased vascular endothelial cadherin and endothelin, suggesting reduced angiogenesis and enhanced vascular integrity. The non-endothelial cell fraction showed decreased expression of VEGFA and fibronectin, without significant difference in the expression of other profibrotic factors. Conclusions: Our findings confirm that adjuvant pre-operative IVB decreased fibronectin and increase endothelin-1 expression without affecting other profibrotic gene expression, uncovering an important interaction between IVB and endothelin-1 that deserves further study.


Asunto(s)
Inhibidores de la Angiogénesis , Bevacizumab , Retinopatía Diabética , Fibrosis , Inyecciones Intravítreas , Factor A de Crecimiento Endotelial Vascular , Vitrectomía , Humanos , Retinopatía Diabética/metabolismo , Retinopatía Diabética/tratamiento farmacológico , Retinopatía Diabética/cirugía , Inhibidores de la Angiogénesis/uso terapéutico , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Bevacizumab/uso terapéutico , Bevacizumab/farmacología , Masculino , Femenino , Persona de Mediana Edad , ARN Mensajero/genética , ARN Mensajero/metabolismo , Neovascularización Retiniana/metabolismo , Neovascularización Retiniana/tratamiento farmacológico , Anciano , Cuidados Preoperatorios , Anticuerpos Monoclonales Humanizados/uso terapéutico , Anticuerpos Monoclonales Humanizados/farmacología
17.
Environ Sci Pollut Res Int ; 31(21): 31135-31148, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38627347

RESUMEN

We have synthesized cobalt ferrite (CoFe2O4) using the sucrose auto-combustion method and subsequently employed the in situ polymerization technique to fabricate ternary composites comprising CoFe2O4, polyaniline (PANI), and multi-walled carbon nanotubes (MWCNTs). In this novel investigation, we explored the influence of varying MWCNTs ratios on these composites' structural, magnetic, thermal, and electrical properties. The crystal structures of the synthesized composites were analyzed using X-ray diffraction (XRD), while Fourier transform infrared (FT-IR) spectroscopy revealed changes in bonding patterns, including the disappearance of ferrite bonds and the emergence of new ones. Transmission electron microscopy (TEM) images illustrated a complete coating of PANI on both MWCNTs and CoFe2O4 particles, resulting in a substantial reduction in magnetization compared to pure CoFe2O4 ferrite due to PANI's nonmagnetic nature. Vibrating sample magnetometer (VSM) measurements confirmed this reduction, indicating a decrease to 7.3 emu.g-1. Thermal analysis demonstrated an enhancement in thermal stability with increasing MWCNTs content, as evidenced by an increase in the temperature equivalent for half decomposition (T50) from 486 to 522 °C for composites with 40% MWCNTs. Moreover, the electrical conductivity showed a corresponding rise with MWCNTs content, increasing from 3.1 × 10-3 Ω-1.cm-1 to 2.2 × 10-2 Ω-1.cm-1, possibly indicating charge transfer from PANI to MWCNTs. To assess practical applications, we investigated the ability of the composite with 40% MWCNTs to remove phenol red (PR) dye from aqueous solutions. Through a systematic study of adsorption parameters and kinetics, we determined optimal conditions for effective dye removal and elucidated the underlying adsorption mechanism. Our results demonstrated the composite's efficiency in dye removal, with a 6.4 mg·g-1 capacity for PR dye, highlighting its potential for environmental remediation efforts.


Asunto(s)
Cobalto , Compuestos Férricos , Nanotubos de Carbono , Cobalto/química , Nanotubos de Carbono/química , Compuestos Férricos/química , Espectroscopía Infrarroja por Transformada de Fourier , Compuestos de Anilina/química , Difracción de Rayos X
18.
J Vis Exp ; (206)2024 Apr 12.
Artículo en Inglés | MEDLINE | ID: mdl-38682901

RESUMEN

Neural stem cells (NSCs) divide and produce newborn neurons in the adult brain through a process called adult neurogenesis. Adult NSCs are primarily quiescent, a reversible cell state where they have exited the cell cycle (G0) yet remain responsive to the environment. In the first step of adult neurogenesis, quiescent NSCs (qNSCs) receive a signal and activate, exiting quiescence and re-entering the cell cycle. Thus, understanding the regulators of NSC quiescence and quiescence exit is critical for future strategies targeting adult neurogenesis. However, our understanding of NSC quiescence is limited by technical constraints in identifying quiescent NSCs (qNSCs) and activated NSCs (aNSCs). This protocol describes a new approach to identify and enrich qNSCs and aNSCs generated in in vitro cultures by imaging NSC autofluorescence. First, this protocol describes how to use a confocal microscope to identify autofluorescent markers of qNSCs and aNSCs to classify NSC activation state using autofluorescence intensity. Second, this protocol describes how to use a fluorescent activated cell sorter (FACS) to classify NSC activation state and enrich samples for qNSCs or aNSCs using autofluorescence intensity. Third, this protocol describes how to use a multiphoton microscope to perform fluorescence lifetime imaging (FLIM) at single-cell resolution, classify NSC activation state, and track the dynamics of quiescent exit using both autofluorescence intensities and fluorescence lifetimes. Thus, this protocol provides a live-cell, label-free, single-cell resolution toolkit for studying NSC quiescence and quiescence exit.


Asunto(s)
Células-Madre Neurales , Células-Madre Neurales/citología , Animales , Ratones , Microscopía Confocal/métodos , Citometría de Flujo/métodos , Imagen Óptica/métodos , Neurogénesis/fisiología
19.
Med Oncol ; 41(5): 117, 2024 Apr 17.
Artículo en Inglés | MEDLINE | ID: mdl-38630325

RESUMEN

Among the most prevalent forms of cancer are breast, lung, colon-rectum, and prostate cancers, and breast cancer is a major global health challenge, contributing to 2.26 million cases with approximately 685,000 deaths worldwide in 2020 alone, typically beginning in the milk ducts or lobules that produce and transport milk during lactation and it is becoming challenging to treat as the tissues are developing resistance, which makes urgent calls for new multitargeted drugs. The multitargeted drug design provides a better solution, simultaneously targeting multiple pathways, even when the drug resists one, it remains effective for others. In this study, we included four crucial proteins that perform signalling, receptor, and regulatory action, namely- NUDIX Hydrolases, Dihydrofolate Reductase, HER2/neu Kinase and EGFR and performed multitargeted molecular docking studies against human-approved drugs using HTVS, SP and extra precise algorithms and filtered the poses with MM\GBSA, suggested a benzodiazepine derivative chlordiazepoxide, used as an anxiolytic agent, can be a multitargeted inhibitor with docking and MM\GBSA score ranging from - 4.628 to - 7.877 and - 18.59 to - 135.86 kcal/mol, respectively, and the most interacted residues were 6ARG, 6GLU, 3TRP, and 3VAL. The QikProp-based ADMET and DFT computations showed the suitability and stability of the drug candidate followed by 100 ns MD simulation in water and MMGBSA on trajectories, resulting in stable performance and many intermolecular interactions to make the complexes stable, which favours that chlordiazepoxide can be a multitargeted breast cancer inhibitor. However, experimental validation is needed before its use.


Asunto(s)
Neoplasias de la Mama , Femenino , Masculino , Humanos , Neoplasias de la Mama/tratamiento farmacológico , Clordiazepóxido , Simulación del Acoplamiento Molecular , Transducción de Señal , Benzodiazepinas , Factores de Transcripción
20.
Cureus ; 16(2): e55105, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38558726

RESUMEN

This narrative review explores the integration of eye-tracking technology in dentistry, aiming to provide a comprehensive overview of its current applications and potential benefits. The review begins by elucidating the fundamental principles of eye tracking, encompassing the various eye-tracking methods and devices commonly used in dental research. It then delves into the role of eye tracking in dental education, where the technology offers a unique perspective on students' visual attention during training and skill acquisition. Moreover, the review examines how eye tracking can aid in assessing and improving dental practitioners' clinical performance, shedding light on areas of improvement and expertise. In patient care, the application of eye-tracking technology offers significant potential. By analyzing patients' gaze patterns and visual focus during dental procedures, clinicians can gain valuable insights into their experiences, identifying sources of anxiety and discomfort. This newfound understanding can pave the way for more patient-centric care and optimized treatment plans. The review also explores the application of eye-tracking technology in designing and evaluating dental interfaces and equipment. By assessing visual ergonomics and usability, researchers can develop user-friendly instruments that enhance dental professionals' workflow and efficiency. However, despite its promise, integrating eye tracking in dentistry is not without challenges. Technical limitations, data analysis complexities, and ethical considerations require careful attention to ensure this technology's ethical and responsible use. In conclusion, this narrative review highlights the growing significance of eye-tracking technology in dentistry. Its applications span dental education, clinical practice, and patient care, holding immense potential to revolutionize how dental procedures are conducted, evaluated, and experienced. Nevertheless, further research and collaboration between dental professionals and eye-tracking experts are necessary to unlock the technology's benefits and ensure its seamless integration into dental practices.

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