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1.
Nucleic Acids Res ; 52(14): 8332-8343, 2024 Aug 12.
Artículo en Inglés | MEDLINE | ID: mdl-38953170

RESUMEN

BRCA2 is an essential tumor suppressor protein involved in promoting faithful repair of DNA lesions. The activity of BRCA2 needs to be tuned precisely to be active when and where it is needed. Here, we quantified the spatio-temporal dynamics of BRCA2 in living cells using aberration-corrected multifocal microscopy (acMFM). Using multicolor imaging to identify DNA damage sites, we were able to quantify its dynamic motion patterns in the nucleus and at DNA damage sites. While a large fraction of BRCA2 molecules localized near DNA damage sites appear immobile, an additional fraction of molecules exhibits subdiffusive motion, providing a potential mechanism to retain an increased number of molecules at DNA lesions. Super-resolution microscopy revealed inhomogeneous localization of BRCA2 relative to other DNA repair factors at sites of DNA damage. This suggests the presence of multiple nanoscale compartments in the chromatin surrounding the DNA lesion, which could play an important role in the contribution of BRCA2 to the regulation of the repair process.


Asunto(s)
Proteína BRCA2 , Daño del ADN , Reparación del ADN , Proteína BRCA2/metabolismo , Proteína BRCA2/genética , Humanos , Cromatina/metabolismo , Núcleo Celular/metabolismo , Núcleo Celular/genética , Línea Celular Tumoral , ADN/metabolismo
2.
Methods Appl Fluoresc ; 10(4)2022 Sep 13.
Artículo en Inglés | MEDLINE | ID: mdl-36063812

RESUMEN

Fluorescence microscopy has widespread applications across biological sciences. It has been routinely used for cell counting, which provides a preliminary diagnostic test for many infectious diseases. Conventional fluorescence microscopes are bulky, expensive, time-intensive and laborious. They often require trained operators to acquire and analyze data. We report a compact automated digital fluorescence microscopy system,i-scope, for cell counting applications. Thei-scopeemploys a total internal reflection fluorescence (TIRF) mode of sample illumination, along with a brightfield mode. It has a magnification of 30X, an optical resolution of ∼0.2µm/pixel and offers sample scanning over 20 mm × 20 mm. A custom-written program enables automated image acquisition and analysis, thereby enhancing ease of operation. It has a compact form-factor and has been developed into a standalone system with a processing unit, screen, and other accessories to offer a portable and economic point-of-care diagnostic solution in low-resource settings. We analysed the performance of the i-scopefor milk somatic cell enumeration and benchmarked it against that of a conventional fluorescence microscope.


Asunto(s)
Microscopía Fluorescente , Microscopía Fluorescente/métodos
3.
Food Chem Toxicol ; 150: 112075, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33617964

RESUMEN

Medicinal or herbal plants are widely used for their many favourable properties and are generally safe without any side effects. Saponins are sugar conjugated natural compounds which possess a multitude of biological activities such as medicinal properties, antimicrobial activity, antiviral activity, etc. Saponin production is a part of the normal growth and development process in a lot of plants and plant extracts such as liquorice and ginseng which are exploited as potential drug sources. Herbal compounds have shown a great potential against a wide variety of infectious agents, including viruses such as the SARS-CoV; these are all-natural products and do not show any adverse side effects. This article reviews the various aspects of saponin biosynthesis and extraction, the need for their integration into more mainstream medicinal therapies and how they could be potentially useful in treating viral diseases such as COVID-19, HIV, HSV, rotavirus etc. The literature presents a close review on the saponin efficacy in targeting mentioned viral diseases that occupy a high mortality rate worldwide. This manuscript indicates the role of saponins as a source of dynamic plant based anti-viral remedies and their various methods for extraction from different sources.


Asunto(s)
Antivirales/aislamiento & purificación , Saponinas/aislamiento & purificación , Antiinfecciosos/farmacología , Antifúngicos/farmacología , Antivirales/farmacología , VIH/efectos de los fármacos , Estructura Molecular , Orthomyxoviridae/efectos de los fármacos , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , SARS-CoV-2/efectos de los fármacos , Saponinas/biosíntesis , Saponinas/química , Saponinas/farmacología
4.
Front Neurosci ; 14: 693, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32742252

RESUMEN

Cyclooxygenases (COX) are enzymes catalyzing arachidonic acid into prostanoids. COX exists in three isoforms: COX-1, 2, and 3. COX-1 and COX-2 have been widely studied in order to explore and understand their involvement in Alzheimer's disease (AD), a progressive neuroinflammatory dementia. COX-2 was traditionally viewed to be expressed only under pathological conditions and to have detrimental effects in AD pathophysiology and neurodegeneration. However, an increasing number of reports point to much more complex roles of COX-2 in AD. Mammalian/mechanistic target of rapamycin (mTOR) has been considered as a hub which integrates multiple signaling cascades, some of which are also involved in AD progression. COX-2 and mTOR are both involved in environmental sensing, growth, and metabolic processes of the cell. They are also known to act in cooperation in many different cancers and thus, their role together in normal cellular functions as well as AD has been explored in this review. Some of the therapeutic approaches targeting COX-2 and mTOR in AD and cancer are also discussed.

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