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Objective: We investigated the impact of vitamin D3 (VD3) supplementation during mouse preantral follicle culture in vitro and the mRNA expression of 25-hydroxylase (CYP2R1), 1-alpha-hydroxylase (CYP27B1), and vitamin D receptor (VDR) in mouse ovarian follicles at different stages.Methods: Preantral follicles were retrieved from 39 BDF1 mice (7–8 weeks old) and then cultured in vitro for 12 days under VD3 supplementation (0, 25, and 50 pg/mL). Follicular development and the final oocyte acquisition were assessed. Preantral follicles were retrieved from 15 other BDF1 mice (7–8 weeks old) and cultured without VD3 supplementation. Three stages of mouse ovarian follicles were obtained (preantral, antral, and ruptured follicles). Total RNA was extracted from the pooled cells (from 20 follicles at each stage), and then reverse transcriptase-polymerase chain reaction was performed to identify mRNA for CYP2R1, CYP27B1, and VDR.Results: The survival of preantral follicles, rates of antrum formation and ruptured follicles (per initiated follicle) and the number of total or mature oocytes were all comparable among the three groups. Both CYP2R1 and CYP27B1 were expressed in antral and ruptured follicles, but not in preantral follicles. VDR was expressed in all three follicular stages.Conclusion: VD3 supplementation in vitro (25 or 50 pg/mL) did not enhance mouse follicular development or final oocyte acquisition. Follicular stage-specific expression of CYP2R1, CYP27B1, and VDR was observed.

Quantification of cellular nanoparticles (NPs) is one of the most important steps in studying NP–cell interactions. Here, a simple method for the estimation of cell-associated silver (Ag) NPs in lung cancer cells (A549) is proposed based on their side scattering (SSC) intensities measured by flow cytometry (FCM). To estimate cellular Ag NPs associated with A549 cells over a broad range of experimental conditions, we measured the normalized SSC intensities (nSSC) of A549 cells treated with Ag NPs with five different core sizes (i.e., 40–200 nm, positively charged) under various exposure conditions that reflect different situations of agglomeration, diffusion, and sedimentation in cell culture media, such as upright and inverted configurations with different media heights. Then, we correlated these nSSC values with the numbers of cellular Ag NPs determined by inductively coupled plasma mass spectrometry (ICPMS) as a well-established cross-validation method. The different core sizes of Ag NPs and the various exposure conditions tested in this study confirmed that the FCM-SSC intensities are highly correlated with their core sizes as well as the amount of cellular Ag NPs over a linear range up to ~80,000 Ag NPs/cell and ~23 nSSC, which is significantly broader than those of previous studies.

Agglomeration of nanoparticles (NPs) in cell culture media can alter their physicochemical properties and colloidal behaviour, which may affect their cellular association and cytotoxicity. Herein, we investigated the impact of agglomeration of silver NPs on their effective dose and cellular association under various in vitro assay conditions. Hydrodynamic sizes, zeta potentials, and effective densities of agglomerates were characterized for two different types of positively charged silver NPs with nominal core sizes of 60 nm and 100 nm. Then, three different dose metrics (administered, effective, and cellular doses) under upright and inverted exposure configurations were compared with the measured physicochemical properties and colloidal behaviours. Changes in the agglomeration characteristics, such as an increment in the hydrodynamic size and a decrement in the effective density, assisted the sedimentation and reduced the diffusion of NPs, leading to a higher effective dose and cellular association in the upright configuration than in the inverted configuration. From the comparison between the two sizes, we have observed that agglomerates of NPs with smaller core diameters could be less stably suspended in the biological media and thus, have higher effective doses and cellular association than agglomerates of NPs with larger core sizes, depending on their agglomeration process. Therefore, the agglomeration characteristics, rather than the intrinsic properties of the bare NPs, seem to have a more important role in determining the effective dose and cellular association of NPs and would be more relevant to understanding cell-nanoparticle interactions and their resultant cytotoxicity.

The incidence of colon cancer has increased remarkably in Korea during the last decade. Fatty acids in cell membranes have been implicated in colorectal carcinogenesis. Few studies have focused on the composition of the phospholipid (PL) fatty acids in the mucosa of colorectal adenomas and cancers. Thus, PL fatty acid patterns were studied in the colorectal mucosa of patients with colorectal adenomatous polyposis (n = 27) and colorectal cancer (n = 8), compared with healthy control subjects (n = 38). Significant increases in the ratios of ω6/ω3 and P/S and unsaturation index were observed in both the colorectal adenomatous polyps and cancerous tissues as compared with the normal tissues of the control subjects. Significantly higher ratios of C18:1/C18:0 and C20:4(n-6)/C20:3(n-6) were also observed in both groups of patients with cancer. In conclusion, although many questions still remain, the significant changes in mucosal PL fatty acid patterns observed in this study suggest that the PL fatty acids may have roles in human colorectal carcinogenesis.

Background and Objectives:Essential hypertension has been accepted as being associated with a loss of the balance between proand anti-oxidations. Essential trace elements, such as Se, Mn, Cu and Zn, participate in various enzyme reactions necessary for the antioxidant defense system of cells. The object of the present study was to evaluate the serum levels of anti-oxidant minerals, which may be related to the pathophysiology of essential hypertension. Subjects and Methods: The subjects were composed of 50 normotensive controls; 25 men and 25 women (14 premenopausal, 11 postmenopausal), and 50 hypertensive patients; 16 men and 32 women (19 premenopausal, 13 postmenopausal). ICP-MS was employed to measure the mineral concentrations. Results:Compared with controls, the serum Se and Zn concentrations in the essential hypertensive patients were elevated (Se 12.78±0.47 vs. 17.85±0.58 μL/dL, p=0.0008; Zn 110.87±5.36 vs. 134.36± 5.95 μL/dL, p=0.0040), whereas those of Mn and Cu were decreased (Mn 0.49±0.02 vs. 0.28±0.03 μL/dL, p=0.0121; Cu 127.84±5.07 vs. 93.88±2.90 μL/dL, p