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1.
Clin Case Rep ; 12(8): e9262, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-39149157

RESUMEN

Small cell lung cancer (SCLC), a neuroendocrine aggressive subtype of lung cancer, is associated with paraneoplastic disorders in about 9% of patients. In this report, we describe a middle-aged man who presented with chronic bowel obstruction caused by chronic intestinal pseudo-obstruction (CIPO) due to SCLC.

2.
Plant Reprod ; 36(1): 1-15, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-35767067

RESUMEN

KEY MESSAGE: In barley (Hordeum vulgare), MTOPVIB is critical for meiotic DSB and accompanied SC and CO formation while dispensable for meiotic bipolar spindle formation. Homologous recombination during meiosis assures genetic variation in offspring. Programmed meiotic DNA double-strand breaks (DSBs) are repaired as crossover (CO) or non-crossover (NCO) during meiotic recombination. The meiotic topoisomerase VI (TopoVI) B subunit (MTOPVIB) plays an essential role in meiotic DSB formation critical for CO-recombination. More recently MTOPVIB has been also shown to play a role in meiotic bipolar spindle formation in rice and maize. Here, we describe a meiotic DSB-defective mutant in barley (Hordeum vulgare L.). CRISPR-associated 9 (Cas9) endonuclease-generated mtopVIB plants show complete sterility due to the absence of meiotic DSB, synaptonemal complex (SC), and CO formation leading to the occurrence of univalents and their unbalanced segregation into aneuploid gametes. In HvmtopVIB plants, we also frequently found the bi-orientation of sister kinetochores in univalents during metaphase I and the precocious separation of sister chromatids during anaphase I. Moreover, the near absence of polyads after meiosis II, suggests that despite being critical for meiotic DSB formation in barley, MTOPVIB seems not to be strictly required for meiotic bipolar spindle formation.


Asunto(s)
Proteínas Arqueales , Hordeum , Hordeum/genética , Hordeum/metabolismo , ADN-Topoisomerasas de Tipo II/genética , ADN-Topoisomerasas de Tipo II/metabolismo , Proteínas Arqueales/genética , Meiosis , ADN , Reparación del ADN
3.
Nucleus ; 13(1): 277-299, 2022 12.
Artículo en Inglés | MEDLINE | ID: mdl-36447428

RESUMEN

Nucleus, chromatin, and chromosome organization studies heavily rely on fluorescence microscopy imaging to elucidate the distribution and abundance of structural and regulatory components. Three-dimensional (3D) image stacks are a source of quantitative data on signal intensity level and distribution and on the type and shape of distribution patterns in space. Their analysis can lead to novel insights that are otherwise missed in qualitative-only analyses. Quantitative image analysis requires specific software and workflows for image rendering, processing, segmentation, setting measurement points and reference frames and exporting target data before further numerical processing and plotting. These tasks often call for the development of customized computational scripts and require an expertise that is not broadly available to the community of experimental biologists. Yet, the increasing accessibility of high- and super-resolution imaging methods fuels the demand for user-friendly image analysis workflows. Here, we provide a compendium of strategies developed by participants of a training school from the COST action INDEPTH to analyze the spatial distribution of nuclear and chromosomal signals from 3D image stacks, acquired by diffraction-limited confocal microscopy and super-resolution microscopy methods (SIM and STED). While the examples make use of one specific commercial software package, the workflows can easily be adapted to concurrent commercial and open-source software. The aim is to encourage biologists lacking custom-script-based expertise to venture into quantitative image analysis and to better exploit the discovery potential of their images.Abbreviations: 3D FISH: three-dimensional fluorescence in situ hybridization; 3D: three-dimensional; ASY1: ASYNAPTIC 1; CC: chromocenters; CO: Crossover; DAPI: 4',6-diamidino-2-phenylindole; DMC1: DNA MEIOTIC RECOMBINASE 1; DSB: Double-Strand Break; FISH: fluorescence in situ hybridization; GFP: GREEN FLUORESCENT PROTEIN; HEI10: HUMAN ENHANCER OF INVASION 10; NCO: Non-Crossover; NE: Nuclear Envelope; Oligo-FISH: oligonucleotide fluorescence in situ hybridization; RNPII: RNA Polymerase II; SC: Synaptonemal Complex; SIM: structured illumination microscopy; ZMM (ZIP: MSH4: MSH5 and MER3 proteins); ZYP1: ZIPPER-LIKE PROTEIN 1.


Asunto(s)
Núcleo Celular , Cromatina , Humanos , Flujo de Trabajo , Hibridación Fluorescente in Situ , Microscopía Fluorescente , Proteínas Fluorescentes Verdes
4.
Microsc Res Tech ; 85(8): 2801-2812, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35411979

RESUMEN

Opuntia is the most diverse and widely distributed drought resistant promising genus of family Cactaceae. The cladodes were utilized to quantify the chemical composition of these plants helpful in lignocellulose conversion and their application towards biofuel production. The present study was aimed to evaluate and compare the taxonomic relationship based on morphology, stem anatomy and palynology of important desert cacti including Opuntia dillenii and Opuntia monacantha. This study also evaluates the potential usefulness of morphological, anatomical and pollen traits using light and scanning electron microscopy. The obtained microcharacters of stem and flowers are considered diagnostic at the generic and specific levels. Some distinguishing morphological features observed were elliptical to obovate cladodes, 1-7 spines per areole and presence of glochidia in O. dillenii. Pollen and stem anatomical characters of the studied taxa are considered highly diagnostic at the generic and species levels. The epidermis has irregular and wavy cells, with straight to sinuate wall pattern and paracytic stomata. Pollen grains appear as pantoporate and prolate spheroidal having reticulate to perforate-reticulate sculpturing while exine semi-tectate to tectate. The taxonomic features studied could be valuable to elaborate and helpful in correctly identification of Opuntia species. The methods of diverse microscopic examination also providing sufficient evidence about the taxonomy of the Opuntia species. RESEARCH HIGHLIGHTS: Description and illustration of desert cacti Opuntia. Morpho-anatomy and palynology were studied with LM and SEM. Highly variation in taxonomic qualitative and quantitative features. Systematic significance based on taxonomic characters was presented.


Asunto(s)
Opuntia , Células Epidérmicas/ultraestructura , Microscopía Electrónica de Rastreo , Polen/ultraestructura
5.
Cancers (Basel) ; 15(1)2022 Dec 30.
Artículo en Inglés | MEDLINE | ID: mdl-36612248

RESUMEN

Phytochemicals possess various intriguing pharmacological properties against diverse pathological conditions. Extensive studies are on-going to understand the structural/functional properties of phytochemicals as well as the molecular mechanisms of their therapeutic function against various disease conditions. Phytochemicals such as curcumin (Cur), genistein (Gen), and tanshinone-IIA (Tan IIA) have multifaceted therapeutic potentials and various efforts are in progress to understand the molecular dynamics of their function with different tools and technologies. Cur is an active lipophilic polyphenol with pleiotropic function, and it has been shown to possess various intriguing properties including antioxidant, anti-inflammatory, anti-microbial, anticancer, and anti-genotoxic properties besides others beneficial properties. Similarly, Gen (an isoflavone) exhibits a wide range of vital functions including antioxidant, anti-inflammatory, pro-apoptotic, anti-proliferative, anti-angiogenic activities etc. In addition, Tan IIA, a lipophilic compound, possesses antioxidant, anti-angiogenic, anti-inflammatory, anticancer activities, and so on. Over the last few decades, the field of proteomics has garnered great momentum mainly attributed to the recent advancement in mass spectrometry (MS) techniques. It is envisaged that the proteomics technology has considerably contributed to the biomedical research endeavors lately. Interestingly, they have also been explored as a reliable approach to understand the molecular intricacies related to phytochemical-based therapeutic interventions. The present review provides an overview of the proteomics studies performed to unravel the underlying molecular intricacies of various phytochemicals such as Cur, Gen, and Tan IIA. This in-depth study will help the researchers in better understanding of the pharmacological potential of the phytochemicals at the proteomics level. Certainly, this review will be highly instrumental in catalyzing the translational shift from phytochemical-based biomedical research to clinical practice in the near future.

6.
Clin Ophthalmol ; 14: 2497-2505, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32904671

RESUMEN

PURPOSE: Glaucoma is one of the most common complications post-penetrating keratoplasty (PK). In this study, we report the Incidence, risk factors and treatment outcomes of intraocular hypertension (IOH) or/and glaucoma post-penetrating keratoplasty (PKG). METHODS: A 5-year descriptive retrospective study, Lebanese patients who underwent PK at Beirut Eye & ENT Specialist Hospital, between 2012 and 2017, were included. Patients with history of glaucoma were excluded. IOH/PKG cases that necessitate treatment were identified and analyzed for the incidence, risk factors and treatment outcomes. RESULTS: A total of 189 eyes of 159 patients were included, with male/female ratio 1.6 and the mean age 47.2±21.3 years. Bullous keratopathy (BK) presented with a high mean age: 70.3 years while ectasia patients were the youngest: 36.5 years. 34.9% of eyes developed high IOP within a mean of 25 months of follow-up distributed between sub-groups of patient with corneal ectasia (22.5%), redo-PK (51.2%), bullous keratopathy (BK) (50%), keratitis (24.9%), and others (dystrophy, trauma …) (21.4%). High IOP developed in 67.4% of the diabetic patients. Visual acuity was less likely to improve in cases developing elevated IOP while postoperative complications were significantly high. In those refractory to medical treatment, trabeculectomy as a glaucoma surgery was effective in lowering the IOP. Combining procedures with PK was not a risk factor for glaucoma. Interrupted sutures and higher number of suturing were associated with increased IOP levels. CONCLUSION: IOH developed in one out of three patients who underwent penetrating keratoplasty. DM, bullous keratopathy, infectious keratitis and redo-PK were highly associated with PKG, whereas high IOP was less likely to develop in cases with keratoconus. Glaucoma is considered a poor prognostic factor in patients post-PK.

7.
Plant Physiol ; 183(4): 1545-1558, 2020 08.
Artículo en Inglés | MEDLINE | ID: mdl-32527734

RESUMEN

Crossovers (COs) ensure accurate chromosome segregation during meiosis while creating novel allelic combinations. Here, we show that allotetraploid (AABB) durum wheat (Triticum turgidum ssp. durum) utilizes two pathways of meiotic recombination. The class I pathway requires MSH4 and MSH5 (MutSγ) to maintain the obligate CO/chiasma and accounts for ∼85% of meiotic COs, whereas the residual ∼15% are consistent with the class II CO pathway. Class I and class II chiasmata are skewed toward the chromosome ends, but class II chiasmata are significantly more distal than class I chiasmata. Chiasma distribution does not reflect the abundance of double-strand breaks, detected by proxy as RAD51 foci at leptotene, but only ∼2.3% of these sites mature into chiasmata. MutSγ maintains the obligate chiasma despite a 5.4-kb deletion in MSH5B rendering it nonfunctional, which occurred early in the evolution of tetraploid wheat and was then domesticated into hexaploid (AABBDD) common wheat (Triticum aestivum), as well as an 8-kb deletion in MSH4D in hexaploid wheat, predicted to create a nonfunctional pseudogene. Stepwise loss of MSH5B and MSH4D following hybridization and whole-genome duplication may have occurred due to gene redundancy (as functional copies of MSH5A, MSH4A, and MSH4B are still present in the tetraploid and MSH5A, MSH5D, MSH4A, and MSH4B are present in the hexaploid) or as an adaptation to modulate recombination in allopolyploid wheat.


Asunto(s)
Triticum/genética , Segregación Cromosómica/genética , Roturas del ADN de Doble Cadena , Meiosis/genética , Meiosis/fisiología , Tetraploidía
8.
Methods Mol Biol ; 2061: 381-402, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31583674

RESUMEN

Here, we describe a protocol for in planta delivery of chemical compounds into meiocytes of different barley genotypes not impacting plant fertility allowing to harvest seeds from treated plants. Compound uptake into meiocytes is assessed by determining 5-ethynyl-2'-deoxyuridine (EdU) incorporation. Similar to EdU, other compounds being soluble in an aqueous solution can be delivered in planta before/during meiosis to decipher their impact on meiosis and meiotic recombination.We give practical advice on how to deliver EdU as compound example (delivery via injection or needle and thread, addition of detergents or surfactants to increase compound uptake), how in planta compound delivery can be established for your plant material under specific growing conditions, how to generate and characterize barley hybrid plants, and how to conduct a meiotic cytological study of (treated) barley plants.


Asunto(s)
Hordeum/genética , Meiosis/genética , Recombinación Genética , Semillas/genética , Alelos , Cromosomas de las Plantas , Análisis Citogenético , Técnica del Anticuerpo Fluorescente , Hibridación Genética , Fenotipo
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