China
Africa
Explore chapters and articles related to this topic
Synthetic Nanoparticles for Anticancer Drugs
Published in Harishkumar Madhyastha, Durgesh Nandini Chauhan, Nanopharmaceuticals in Regenerative Medicine, 2022
The nanoparticle surface is immobilised by the LL-37 peptide via an oxidation reaction between the primary amine and the carboxyl peptide group of magnetic NPs. To obtain magnetic NPs that function with ceragenin, ethanol and a 25% glutaricdialdehyde solution were added to the core part of the structure. Finally, magnetic NPs with the terminal aldehyde group were resuspended in ethanol containing the synthetic ceragenin analogue to synthesise CSA-13. After synthesis, the precipitated solvent was washed three times with C2H5OH salt solution and a phosphate bufferand then placed at 37°C. To prepare fluorescent nanosystems, magnetic NPs with aldehydes were mixed with propidium iodide (Shukla et al. 2005; Geetha et al. 2013). Magnetic NPs were coated with cathelicidin LL-37. Finally, they were labelled with fluorescein isothiocyanate (Leszczynska et al. 2013; Sorensen et al. 2001).
Asthma, Eosinophils, and Interleukin-5
Published in Gerald J. Gleich, A. Barry Kay, Eosinophils in Allergy and Inflammation, 2019
A. Barry Kay, Qutayba Hamid, Douglas S. Robinson, Andrew M. Bentley, Anne Tsicopoulos, Sun Ying, Redwan Moqbel, Christopher J. Corrigan, Stephen R. Durham
To examine whether mRNA for IL-5 was expressed by T cells, cytocentrifuge preparations of BAL cells from three subjects with asthma were fixed in 4% paraformaldehyde and washed in 15% sucrose in phosphate-buffered saline (20). Cells were incubated simultaneously with IL-5 cRNA probes labeled with uridine triphosphate-biotin and with monoclonal antibody to CD3 directly conjugated to fluorescein isothiocyanate. The conditions for in situ hybridization were as previously described (21), and positive hybridization of probe to cytokine mRNA was detected using streptavidin-Texas red staining. Controls were IgGl-fluorescein isothiocyanate with sense probes and RNase pretreatment. Cells were quantified by fluorescence microscopy, and the percentages of cells expressing both CD3 and cytokine mRNA as well as the percentages of singly labeled cells were evaluated by counting at least 200 positive cells.
Estrogen Receptors and Hormone Responsiveness in Serially Transplanted Mammary Tumors in Rats*
Published in Louis P. Pertschuk, Sin Hang Lee, Localization of Putative Steroid Receptors, 2019
Chung Lee, Charmayne Jesik, Marayart Mangkornkanok, Julia Sensibar, Sin Hang Lee
The experimental sections were air dried for 1 hr in a refrigerator kept at 5°C after they were cut. They were then rehydrated with 2% bovine serum albumin (BSA) for 2 min, then carefully drained. The fluorescent conjugate, 17-beta-estradiol-6-(0)-carboxymenthyl)oxime-BSA-fluorescein isothiocyanate (E2-BSA-FITC), was added to each section so that they were completely covered, and the slides were placed in a darkened humidified chamber for 2 hr. The slides were removed and the sections were drained of excess conjugate; they were washed by inverting the slides in phosphate-buffered saline (PBS) twice for 30 min each time. The sections were then mounted in buffered glycerol and examined with a Leitz fluorescent microscope. The microscope was equipped with an HB-250 mercury vapor lamp, BG-12 exciter filter, and 525 barrier filter.
Topical delivery of Bruton’s tyrosine kinase inhibitor and curcumin-loaded nanostructured lipid carrier gel: Repurposing strategy for the psoriasis management
Published in Pharmaceutical Development and Technology, 2022
Harsha Jain, Geetanjali Devabattula, Aditi Bhat, Harshita Dalvi, Nagarjun Rangaraj, Chandraiah Godugu, Saurabh Srivastava
The confocal laser scanning microscopy (CLSM) was used to check the skin permeation of prepared formulations. For distribution of prepared NLC loaded animals were kept in central animal house facility at NIPER-Hyderabad under monitored conditions (12 h light/12 h of darkness at 22 °C with 50% relative humidity (RH)) for a week to acclimatize. The distribution of prepared formulation into skin layers was examined using shaved mice. The Fluorescein isothiocyanate (FITC) (0.25% w/v) loaded NLC formulations were prepared as per the procedure described in Section 2.4. FITC loaded NLC and plain drug were applied uniformly to the skin and kept for 6 h in dark. The animals were sacrificed after the completion of study, and the skin sections were collected and kept at 4 °C in 30% sucrose solution. The experiment was executed in psoriatic and healthy animal. Sections were cut with the help of cryotome (Leica Biosystems, Germany). The sections were mounted on slides and examined using a microscope (Nikon Corporation, Japan) (Jain et al. 2016).
Cancer Chemopreventive Properties of Sulfated Enterolobium cyclocarpum Extract
Published in Nutrition and Cancer, 2021
Amira M. Gamal-Eldeen, Hassan Amer, Amani A Alrehaili, Ahmed Saleh, Abd El-Rahman Al Ghamdi, Nahed M Hawsawi, Asma Salman, Bassem M. Raafat
To measure the macrophage proliferation index, RAW 264.7 macrophages (0.5 × 105 cells/well), were treated with SEC (0–40 µg/ml) and cultured in phenol red-free RPMI for 20 h at 37ºC, in a humidified 5% CO2. After incubation, cells were submitted to 3-[4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay (24), which is based on the cleavage of the tetrazolium salt by mitochondrial dehydrogenases in viable cells. The accumulation of nitrite, an indicator of NO synthesis, was measured by a Griess reagent in RAW 264.7 cells (25). Macrophages were pre-incubated for 24 h with bacterial lipopolysaccharide (LPS, 1 µg/ml) with or without SEC (10 µg/ml). The nitrite content was normalized to the cellular protein content. The effect of SEC on the binding affinity of fluorescein isothiocyanate (FITC)-conjugated LPS to RAW 264.7 cells was monitored (26). Cells were pre-incubated with or without SEC (10 µg/ml) for 30 min at 37 °C in RPMI 1640 containing 10% FBS, as a source of CD14 and LPS-binding protein. After washing, the cells were incubated for 1 h with FITC-conjugated LPS (200 ng/ml) and the binding of FITC-LPS was analyzed by a microplate fluorescence reader (FluoStarOptima, BMG, USA).
A novel dendrimer-based complex co-modified with cyclic RGD hexapeptide and penetratin for noninvasive targeting and penetration of the ocular posterior segment
Published in Drug Delivery, 2019
Xiucheng Yang, Lihua Wang, Lin Li, Meishan Han, Shengnan Tang, Tengteng Wang, Junping Han, Xiaoyan He, Xiuting He, Aiping Wang, Kaoxiang Sun
To maximize both targeting of the CNV and crossing of the ocular physiological barriers, in the current study, RGD and PEN co-modified PEGylated PAMAM was successfully constructed. It was possible to adjust the size by changing the molecular weight and grafting density of the PEG. Fluorescein isothiocyanate (FITC) was used as a fluorescent probe for cell and animal imaging experiments and it was linked to PAMAM. The cytotoxicity of NCs in human conjunctival epithelial cells (NHCs), human corneal epithelial cells (HCECs), and human umbilical vein endothelial cells (HUVECs) was measured using a MTT assay. Because of the wide application of HUVECs as vascular endothelial cells in investigations of pathologic neovascularization, HUVECs were applied as a cellular model to validate the targeting of the NCs. The in vitro permeability of the NCs was evaluated in NHCs and HCECs. Furthermore, frozen sections of mouse eyes were used to evaluate the in vivo distribution of NCs. The penetration and targeting of the NCs were studied to identify their availability and feasibility for use in the treatment of ocular posterior segment diseases. We hope that the current research will contribute to the development of a noninvasive CNV eyedrop treatment.