5 research outputs found
Torpedo maculopathy: A primary choroidal capillary abnormality?
A 26-year-old healthy male patient's fundus revealed findings consistent with torpedo maculopathy. Swept-source optical coherence tomography (OCT) showed a dome-shaped elevation of the retina at the level of ellipsoid zone. On OCT angiography segmented at the level of the choriocapillaris, a cluster of convoluted fine vessels was seen, and further, deeper scans of the larger choroidal vessels showed a slower flow. From these observations along with the embryological correlation of choriocapillaris development, a possibility of an abnormality preventing proper fenestration of the choriocapillaris along the horizontal raphe being responsible for this anomaly is suggested
Rapid flow cytometry based cytotoxicity assay for evaluation of NK cell function
983-988Assessment of natural killer cells (NK-cell)
cytotoxicity is used not only in research settings but is also important in
diagnosis of various diseases. NK-cell cytotoxicity assays are based on
measurement of target cells killed by cytotoxic cells analyzed either by
chromium (51Cr) release assay or flow cytometry. Both these methods
use peripheral blood mononuclear cells (PBMC) or pure NK-cell population and
hence require large volume of blood sample which is difficult to obtain in
pediatric patients and patients with cytopenia. Hence, a flow cytometric assay
was designed to determine NK cell activity using whole blood, eliminating the
need for isolation of PBMCs or pure NK cells. This assay is based on a dual
fluorescent staining of target cells (K562 cell line). The DIOC18 dye labeled
K562 cells are incubated with whole blood and then counterstained with 7-AAD
enabling the measurement of dead target cell and then percent cytotoxicity is
calculated. This study compared the NK cell cytotoxicity using PBMC and whole
blood in clinically relevant samples. There was no significant difference
between two assays in the measurement of lytic activity or in reproducibility
in the repeated samplings of healthy individuals. The whole blood assay
required less volume of blood and also less processing time as compared to PBMC
assay. It was also validated by testing patients diagnosed with familial
hemophagocytic lymphohistiocytosis expected to have low NK-cell activity. This
assay is rapid, sensitive and reproducible and requires significantly less
volume of blood which is important for clinical evaluation of NK-cell function
Asialoglycoprotein receptor targeted delivery of doxorubicin nanoparticles for hepatocellular carcinoma
<p>We report asialoglycoprotein receptor (ASGPR)-targeted doxorubicin hydrochloride (Dox) nanoparticles (NPs) for hepatocellular carcinoma (HCC). Polyethylene sebacate (PES)-Gantrez® AN 119 Dox NPs of average size 220 nm with PDI < 0.62 and ∼20% Dox loading were prepared by modified nanoprecipitation. ASGPR ligands, pullulan (Pul), arabinogalactan (AGn), and the combination (Pul-AGn), were anchored by adsorption. Ligand anchoring enabled high liver uptake with a remarkable hepatocyte:nonparenchymal cell ratio of 85:15. Furthermore, Pul-AGn NPs exhibited an additive effect implying incredibly high hepatocyte accumulation. Galactose-mediated competitive inhibition confirmed ASGPR-mediated uptake of ligand-anchored NPs in HepG2 cell lines. Subacute toxicity in rats confirmed the safety of the NP groups. However, histopathological evaluation suggested mild renal toxicity of AGn. Pul NPs revealed sustained reduction in tumor volume in PLC/PRF/5 liver tumor-bearing Nod/Scid mice up to 46 days. Extensive tumor necrosis, reduced collagen content, reduction in the HCC biomarker serum α-fetoprotein (<i>p</i> < 0.05), a mitotic index of 1.135 (day 46), and tumor treated/tumor control (T/C) values of <0.42 signified superior efficacy of Pul NPs. Furthermore, weight gain in the NP groups, and no histopathological alterations indicated that they were well tolerated by the mice. The high efficacy coupled with greater safety portrayed Pul Dox NPs as a promising nanocarrier for improved therapy of HCC.</p