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Bio-protocol Sep 2023Administration of substances into neonatal mice is required for early treatment with pre-clinical therapeutics, delivery of recombination-inducing substances, and dosing...
Administration of substances into neonatal mice is required for early treatment with pre-clinical therapeutics, delivery of recombination-inducing substances, and dosing with viruses or toxins, amongst other things. Several injection routes into mouse pups are possible, including intravenous and intracerebroventricular, each with their own advantages and limitations. Here, we describe a simple and rapid protocol for the intraperitoneal injection of neonatal mice for systemic dosing. By detaching a 30-gauge needle from its plastic hub and inserting it into polyethylene tubing attached to a Hamilton syringe, small volumes (1-10 μL) can be accurately injected into the peritoneal cavity of pups aged 1-5 days old. The procedure can be completed within a few minutes, is generally safe and well tolerated by both pups and parents, and can be used in combination with alternative administration routes. Key features • This protocol provides a simple description to rapidly and efficiently inject mouse pups aged 1-5 days for systemic dosing. • Allows treatment of neonatal mice with substances such as viruses and compounds for research across disciplines.
PubMed: 37753468
DOI: 10.21769/BioProtoc.4826 -
Biomedicine & Pharmacotherapy =... Mar 2024Vitreous replacement is a commonly employed method for treating a range of ocular diseases, including posterior vitreous detachment, complex retinal detachment, diabetic... (Review)
Review
Vitreous replacement is a commonly employed method for treating a range of ocular diseases, including posterior vitreous detachment, complex retinal detachment, diabetic retinopathy, macular hole, and ocular trauma. Various clinical substitutes for vitreous include air, expandable gas, silicone oil, heavy silicone oil, and balanced salt solution. However, these substitutes have drawbacks such as short retention time, cytotoxicity, high intraocular pressure, and the formation of cataracts, rendering them unsuitable for long-term treatment. Polymeric hydrogels possess the potential to serve as ideal vitreous substitutes due to their structure-mimicking to natural vitreous and adjustable mechanical properties. Replacement with hydrogels as the tamponade can help maintain the shape of the eyeball, apply pressure to the detached retina, and ensure the metabolic transport of substances without impairing vision. This literature review examines the required properties of artificial vitreous, including the optical properties, rheological properties, expansive force action, and physiological and biochemical functions of chemically and physically crosslinked hydrogels. The strategies for enhancing the biocompatibility and injectability of hydrogels are also summarized and discussed. From a clinical ophthalmology perspective, this paper presents the latest developments in vitreous replacement, providing clinicians with a comprehensive understanding of hydrogel clinical applications, which offers guidance for future design directions and methodologies for hydrogel development.
Topics: Humans; Hydrogels; Eye; Cataract; Diabetic Retinopathy; Ophthalmology; Polymers
PubMed: 38306844
DOI: 10.1016/j.biopha.2024.116154 -
Heliyon Mar 2024Extrachromosomal DNAs (ecDNAs) are a pervasive feature found in cancer and contain oncogenes and their corresponding regulatory elements. Their unique structural... (Review)
Review
Extrachromosomal DNAs (ecDNAs) are a pervasive feature found in cancer and contain oncogenes and their corresponding regulatory elements. Their unique structural properties allow a rapid amplification of oncogenes and alter chromatin accessibility, leading to tumorigenesis and malignant development. The uneven segregation of ecDNA during cell division enhances intercellular genetic heterogeneity, which contributes to tumor evolution that might trigger drug resistance and chemotherapy tolerance. In addition, ecDNA has the ability to integrate into or detach from chromosomal DNA, such progress results into structural alterations and genomic rearrangements within cancer cells. Recent advances in multi-omics analysis revealing the genomic and epigenetic characteristics of ecDNA are anticipated to make valuable contributions to the development of precision cancer therapy. Herein, we conclud the mechanisms of ecDNA generation and the homeostasis of its dynamic structure. In addition to the latest techniques in ecDNA research including multi-omics analysis and biochemical validation methods, we also discuss the role of ecDNA in tumor development and treatment, especially in drug resistance, and future challenges of ecDNA in cancer therapy.
PubMed: 38545177
DOI: 10.1016/j.heliyon.2024.e27733 -
Cell Death & Disease Dec 2023Photoreceptor cell death and immune cell infiltration are two major events that contribute to retinal degeneration. However, the relationship between these two events...
Photoreceptor cell death and immune cell infiltration are two major events that contribute to retinal degeneration. However, the relationship between these two events has not been well delineated, primarily because of an inadequate understanding of the immunological processes involved in photoreceptor degeneration, especially that of peripheral leukocytes that infiltrate the subretinal space and retinal tissues. In this work, we characterized the role of leukocyte infiltration within the detached retina. We observed that CD45 CD11b Ly6G neutrophils and CD45 CD11b Ly6G Ly6C monocytes are the predominant peripheral immune cell populations that infiltrate the retinal and subretinal space after detachment. Selective depletion of monocytes or neutrophils using cell-specific targeting is neuroprotective for photoreceptors. These results indicate that peripheral innate immune cells contribute to photoreceptor degeneration, and targeting these immune cell populations could be therapeutic during retinal detachment.
Topics: Humans; Animals; Retinal Detachment; Monocytes; Neutrophils; Photoreceptor Cells; Retina; Retinal Degeneration; Photoreceptor Cells, Vertebrate; Disease Models, Animal
PubMed: 38102109
DOI: 10.1038/s41419-023-06350-6 -
Ophthalmology. Retina Oct 2023To compare outcomes of scleral buckle (SB), pars plana vitrectomy (PPV), and combined PPV-SB to treat rhegmatogenous retinal detachments (RRDs) with inferior retinal... (Meta-Analysis)
Meta-Analysis Review
TOPIC
To compare outcomes of scleral buckle (SB), pars plana vitrectomy (PPV), and combined PPV-SB to treat rhegmatogenous retinal detachments (RRDs) with inferior retinal breaks (IRBs).
CLINICAL RELEVANCE
Rhegmatogenous retinal detachments with IRBs are not uncommon; their management is challenging with higher risk of failure. There is no consensus about their treatment, specifically whether SB, PPV, or PPV-SB should be performed.
METHODS
Systematic review and meta-analysis. Randomized controlled trials, case-control, and prospective/retrospective series (if n > 50) in English were eligible. Medline, Embase, and Cochrane databases were searched up to January 23, 2023. Standard systematic review methods were followed. The following outcomes at 3 (± 1) and 12 (± 3) months were evaluated: number of eyes with retinal reattachment after ≥ 1 surgeries, change in best-corrected visual acuity from preoperative to postoperative levels, and number of eyes with improvement of > 10 and > 15 ETDRS letters after surgery. Authors of eligible studies were asked for individual participant data (IPD) and IPD meta-analysis was undertaken. Risk of bias was assessed using National Institutes of Health study quality assessment tools. This study was registered prospectively in PROSPERO (CRD42019145626).
RESULTS
A total of 542 studies were identified: 15 were eligible and included and 60% were retrospective. Individual participant data was obtained from 8 studies (1017 eyes). Given that only 26 patients had received SB alone, these data were not considered in the analysis. There was no evidence for differences between treatment groups (PPV versus PPV-SB) in the probability of having a flat retina at 3 or 12 months postoperatively after 1 (P = 0.067; odds ratio [OR], 0.47; P = 0.408; OR 2.55; respectively) or > 1 (OR, 0.54; P = 0.21; OR, 0.89; P = 0.926; respectively) surgery. Pars plana vitrectomy-SB showed less improvement in vision postoperatively at 3 months (estimate, 0.18; 95% confidence interval, 0.01-0.35; P = 0.044), but this difference was no longer observed at 12 months (estimate, -0.07; 95% confidence interval, -0.27, 0.13; P = 0.479).
CONCLUSION
Available evidence suggests a lack of benefit of adding SB to PPV to treat RRDs with IRBs. Evidence, however, comes mainly from retrospective series and, thus, despite the large number of eyes included, should be interpreted with caution. Further research is needed.
FINANCIAL DISCLOSURE(S)
The author(s) have no proprietary or commercial interest in any materials discussed in this article.
Topics: Humans; Retinal Detachment; Vitrectomy; Retrospective Studies; Prospective Studies; Visual Acuity; Retinal Perforations
PubMed: 37187441
DOI: 10.1016/j.oret.2023.05.006 -
Ceska a Slovenska Oftalmologie :... 2024To summarize the history and current trends in the use of scleral grafts in ophthalmology. (Review)
Review
AIM
To summarize the history and current trends in the use of scleral grafts in ophthalmology.
MATERIALS AND METHODS
We conducted a review of the literature through the MEDLINE and Cochrane Library databases. The search terms were "sclera", "graft", and "surgery". The search resulted in 1596 articles, of which we evaluated 192 as relevant. The relevant articles were sorted chronologically and according to the method of using scleral grafts, which enabled the development of a review article.
RESULTS
The sclera has been routinely used in ophthalmology since the 1950s in many different indications. Some of these indications have become practically obsolete over time (for example, use in the surgical management of retinal detachment), but a large number still find application today (especially use in glaucoma or oculoplastic surgery, or as a patch for a defect in the sclera or cornea).
CONCLUSION
Even though allogeneic sclera is currently used less frequently in ophthalmology compared to other tissue banking products and the range of its indications has partially narrowed, it remains a useful material due to its availability and properties.
Topics: Humans; Ophthalmology; Sclera; Glaucoma; Retinal Detachment; Cornea
PubMed: 38413226
DOI: 10.31348/2024/11 -
Frontiers in Cell and Developmental... 2023Detachment from the extracellular matrix (ECM) is the first step of the metastatic cascade. It is a regulated process involving interaction between tumor cells and...
Detachment from the extracellular matrix (ECM) is the first step of the metastatic cascade. It is a regulated process involving interaction between tumor cells and tumor microenvironment (TME). Iron is a key micronutrient within the TME. Here, we explored the role of iron in the ability of ovarian cancer cells to successfully detach from the ECM. HEY and PEO1 ovarian cancer cells were grown in 3D conditions. To mimic an iron rich TME, culture media were supplemented with 100 μM Fe. Cell mortality was evaluated by cytofluorimetric assay. The invasive potential of tumor spheroids was performed in Matrigel and documented with images and time-lapses. Iron metabolism was assessed by analyzing the expression of CD71 and FtH1, and by quantifying the intracellular labile iron pool (LIP) through Calcein-AM cytofluorimetric assay. Ferroptosis was assessed by quantifying mitochondrial reactive oxygen species (ROS) and lipid peroxidation through MitoSOX and BODIPY-C11 cytofluorimetric assays, respectively. Ferroptosis markers GPX4 and VDAC2 were measured by Western blot. knockdown was performed by using siRNA. To generate spheroids, HEY and PEO1 cells prevent LIP accumulation by upregulating FtH1. 3D HEY moderately increases FtH1, and LIP is only slightly reduced. 3D PEO1upregulate FtH1 and LIP results significantly diminished. HEY tumor spheroids prevent iron import downregulating CD71, while PEO1 cells strongly enhance it. Intracellular ROS drop down during the 2D to 3D transition in both cell lines, but more significantly in PEO1 cells. Upon iron supplementation, PEO1 cells continue to enhance CD71 and FtH1 without accumulating the LIP and ROS and do not undergo ferroptosis. HEY, instead, accumulate LIP, undergo ferroptosis and attenuate their sphere-forming ability and invasiveness. knockdown significantly reduces the generation of PEO1 tumor spheroids, although without sensitizing them to ferroptosis. Iron metabolism reprogramming is a key event in the tumor spheroid generation of ovarian cancer cells. An iron-rich environment impairs the sphere-forming ability and causes cell death only in ferroptosis sensitive cells. A better understanding of ferroptosis sensitivity could be useful to develop effective treatments to kill ECM-detached ovarian cancer cells.
PubMed: 38033861
DOI: 10.3389/fcell.2023.1272667 -
Nature Communications Oct 2023Transglutaminase 3 (TG3), the autoantigen of dermatitis herpetiformis (DH), is a calcium dependent enzyme that targets glutamine residues in polypeptides for either...
Transglutaminase 3 (TG3), the autoantigen of dermatitis herpetiformis (DH), is a calcium dependent enzyme that targets glutamine residues in polypeptides for either transamidation or deamidation modifications. To become catalytically active TG3 requires proteolytic cleavage between the core domain and two C-terminal β-barrels (C1C2). Here, we report four X-ray crystal structures representing inactive and active conformations of human TG3 in complex with a TG3-specific Fab fragment of a DH patient derived antibody. We demonstrate that cleaved TG3, upon binding of a substrate-mimicking inhibitor, undergoes a large conformational change as a β-sheet in the catalytic core domain moves and C1C2 detaches. The unique enzyme-substrate conformation of TG3 without C1C2 is recognized by DH autoantibodies. The findings support a model where B-cell receptors of TG3-specific B cells bind and internalize TG3-gluten enzyme-substrate complexes thereby facilitating gluten-antigen presentation, T-cell help and autoantibody production.
Topics: Humans; Celiac Disease; Autoantibodies; Dermatitis Herpetiformis; Transglutaminases; Immunoglobulin A; Glutens
PubMed: 37798283
DOI: 10.1038/s41467-023-42004-z -
Ophthalmology and Therapy Oct 2023Retinopathy of prematurity (ROP), a leading cause of childhood blindness, has historically been associated with blindness from overgrowth of blood vessels from the... (Review)
Review
Retinopathy of prematurity (ROP), a leading cause of childhood blindness, has historically been associated with blindness from overgrowth of blood vessels from the retina into the vitreous that lead to complex retinal detachments. Our understanding of ROP has evolved with the survival of extremely low-birthweight infants and includes not only overgrowth of blood vessels, but also insufficient developmental retinal vascular growth in early phases of the disease. Our current treatments of ROP have focused on methods to improve perinatal and prenatal care, reduce premature birth, and prevent early phases of ROP. Nonetheless, addressing vasoproliferation in treatment-warranted eyes remains the mainstay of management. Two main treatment strategies co-exist today: laser treatment, which has been the standard of care since the 1990s, and anti-VEGF injections, which have been used since early reports in 2007 (Travassos et al. in Ophthalmic Surg Lasers Imaging, 38:233-237, https://doi.org/10.3928/15428877-20070501-09 , 2007, Shah et al. in Indian J Ophthalmol 55:75-76, https://doi.org/10.4103/0301-4738.29505 , 2007, Quiroz-Mercado et al. in Semin Ophthalmol 22:109-125, https://doi.org/10.1080/08820530701420082 , 2007).
PubMed: 37337024
DOI: 10.1007/s40123-023-00744-7