Fabrication and assessment of milk phospholipid-complexed antioxidant phytosomes with vitamin C and E: A comparison with liposomes.

Evidences have shown that phytosome assemblies are novel drug delivery system. However, studies of phytosomes in food applications are scarce. The characteristics of milk phospholipid assemblies and their functionality in terms of in vitro digestibility and bioavailability of encapsulated nutrients (ascorbic acid and α-tocopherol) were studied. The phytosomes were fabricated using ethanolic evaporation technique. Spectral analysis revealed that polar parts of phospholipids formed hydrogen bonds with ascorbic acid hydroxyl groups, further, incorporating ascorbic acid or α-tocopherol into the phospholipid assembly changed the chemical conformation of the complexes. Phospholipid-ascorbic acid phytosomes yielded an optimal complexing index of 98.52 ± 0.03% at a molar ratio of 1:1. Phytosomes exhibited good biocompatibility on intestinal epithelial cells. The cellular uptake of ascorbic acid was 29.06 ± 1.18% for phytosomes. It was higher than that for liposomes (24.14 ± 0.60%) and for ascorbic acid aqueous solution (1.17 ± 0.70%).

Time-of-flight resolved light field fluctuations reveal deep human tissue physiology.

Red blood cells (RBCs) transport oxygen to tissues and remove carbon dioxide. Diffuse optical flowmetry (DOF) assesses deep tissue RBC dynamics by measuring coherent fluctuations of multiply scattered near-infrared light intensity. While classical DOF measurements empirically correlate with blood flow, they remain far-removed from light scattering physics and difficult to interpret in layered media. To advance DOF measurements closer to the physics, here we introduce an interferometric technique, surmounting challenges of bulk motion to apply it in awake humans. We reveal two measurement dimensions: optical phase, and time-of-flight (TOF), the latter with 22 picosecond resolution. With this multidimensional data, we directly confirm the unordered, or Brownian, nature of optically probed RBC dynamics typically assumed in classical DOF. We illustrate how incorrect absorption assumptions, anisotropic RBC scattering, and layered tissues may confound classical DOF. By comparison, our direct method enables accurate and comprehensive assessment of blood flow dynamics in humans.

Preparation of novel phospholipid-based sonocomplexes for improved intestinal permeability of rosuvastatin: In vitro characterization, dynamic simulation, Caco-2 cell line permeation and in vivo assessment studies.

The study aimed to fabricate innovative drug-phospholipid complexes termed "sonocomplexes" adopting ultrasound irradiation to increase the liposolubility and to enhance the intestinal absorption of rosuvastatin as a model drug for BCS class III active pharmaceutical ingredients (APIs). A 22 full factorial design was fashioned to investigate the influence of phosphatidylcholine content in the phospholipid (∼30 and 60%) and molar ratio of phospholipid to rosuvastatin (1:1 and 2:1) on physicochemical properties of sonocomplexes. In comparison to pure drug, sonocomplexes showed a minimum of about 2 folds and a maximum of about 15 folds increase in lipophilicity (expressed in terms of partition coefficient, P). Results of molecular docking, dynamic simulations, Fourier transform infrared (FTIR) spectroscopy and differential scanning calorimetry (DSC) confirmed the strong interactions between rosuvastatin and the phospholipid via hydrogen bonding interaction, van der Waals forces and hydrophobic interaction. The complexation efficiency reached around 99% and transmission electron microscopy (TEM) of the aqueous dispersion of the optimal sonocomplex showed spherical nanosized vesicles. The optimal sonocomplex showed significantly superior Caco-2 cells permeability and markedly better oral bioavailability compared to the pure drug. In summary, sonocomplexes can be considered as effective approach for enhancing the liposolubility and consequently the intestinal permeability of BCS class III drugs.

Reliability, repeatability, and reproducibility of pulmonary transit time assessment by contrast enhanced echocardiography.

BACKGROUND: The aim of this study is to investigate the inter and intra-rater reliability, repeatability, and reproducibility of pulmonary transit time (PTT) measurement in patients using contrast enhanced ultrasound (CEUS), as an indirect measure of preload and left ventricular function. METHODS: Mean transit times (MTT) were measured by drawing a region of interest (ROI) in right and left cardiac ventricle in the CEUS loops. Acoustic intensity dilution curves were obtained from the ROIs. MTTs were calculated by applying model-based fitting on the dilution curves. PTT was calculated as the difference of the MTTs. Eight raters with different levels of experience measured the PTT (time moment 1) and repeated the measurement within a week (time moment 2). Reliability and agreement were assessed using intra-class correlations (ICC) and Bland-Altman analysis. Repeatability was tested by estimating the variance of means (ANOVA) of three injections in each patient at different doses. Reproducibility was tested by the ICC of the two time moments. RESULTS: Fifteen patients with heart failure were included. The mean PTT was 11.8 ± 3.1 s at time moment 1 and 11.7 ± 2.9 s at time moment 2. The inter-rater reliability for PTT was excellent (ICC = 0.94). The intra-rater reliability per rater was between 0.81-0.99. Bland-Altman analysis revealed a bias of 0.10 s within the rater groups. Reproducibility for PTT showed an ICC = 0.94 between the two time moments. ANOVA showed no significant difference between the means of the three different doses F = 0.048 (P = 0.95). The mean and standard deviation for PTT estimates at three different doses was 11.6 ± 3.3 s. CONCLUSIONS: PTT estimation using CEUS shows a high inter- and intra-rater reliability, repeatability at three different doses, and reproducibility by ROI drawing. This makes the minimally invasive PTT measurement using contrast echocardiography ready for clinical evaluation in patients with heart failure and for preload estimation.

Intrahepatic transit time predicts liver fibrosis in patients with chronic hepatitis B: quantitative assessment with contrast-enhanced ultrasonography.

We investigated the use of contrast-enhanced ultrasonography (CEUS) with quantitative measurements to assess the stages of liver fibrosis in patients with chronic hepatitis B. One-hundred twenty-two patients with chronic hepatitis B were divided into three groups according to the Scheuer scoring system pathologically and according to clinical evidence: mild fibrosis (S0 and S1, n = 36); moderate fibrosis (S2 and S3, n = 24); and cirrhosis (S4 and clinically typical cirrhosis, n = 62). CEUS of hepatic vessels and parenchyma was performed using the Cadence contrast pulse sequencing technique, with an intravenous bolus injection of a contrast agent (SonoVue). Real-time CEUS imaging of the liver was recorded and analyzed offline. Contrast arrival time, baseline, and peak intensity in the hepatic artery, portal vein, right hepatic vein, and liver parenchyma were used to calculate intrahepatic transit times, hepatic artery to hepatic vein transit time (HA-HVTT) and portal vein to hepatic vein transit time (PV-HVTT), as well as increased signal intensity (ISI). The correlations between these quantitative parameters and the stages of fibrosis were analyzed using Spearman rank correlation coefficients. HA-HVTT and PV-HVTT were shortened gradually with the progression of liver fibrosis. PV-HVTT was statistically significant differences existed between the two paired groups (mild vs. moderate vs. cirrhosis groups, p < 0.001), whereas HA-HVTT was changed significantly between mild and moderate or cirrhosis groups (p < 0.001). HA-HVTT and PV-HVTT changes were significantly correlated with liver fibrosis severity (r = -0.5930, p < 0.001; r = -0.8215, p < 0.001). Area under receiver operating characteristic curves for HA-HVTT and PV-HVTT were 0.891 +/- 0.034 and 0.955 +/- 0.020 at fibrosis scores >or=S2, and 0.785 +/- 0.040 and 0.946 +/- 0.018 at fibrosis score >or=S4, respectively. ISI values in the portal vein and liver parenchyma decreased with the severity of fibrosis. This study demonstrated that hepatic CEUS with quantitative measurements of intrahepatic transit time reflected the severity of liver fibrosis. The real-time CEUS imaging with use of software-based quantitative analysis could provide reliable information of hepatic hemodynamic changes to noninvasively assess the severity of liver fibrosis in patients with chronic hepatitis B.

Characterization of focal liver lesions by means of assessment of hepatic transit time with contrast-enhanced US.

PURPOSE: To assess whether hepatic transit times (HTTs), as measured with contrast material-enhanced ultrasonography (US), can help predict the nature of focal liver lesions. MATERIALS AND METHODS: The study was approved by the local institutional ethics committee, with written informed patient consent. A total of 402 patients were enrolled in the study. HTT, the time between the appearance of the microbubble contrast agent in the hepatic artery and its appearance in the hepatic vein, was measured in the contrast pulse sequencing mode after injection of a sulphur hexafluoride microbubble US contrast agent. Logistic regression was used to identify factors indicative of the malignant or nonmalignant status of focal liver lesions. Receiver operating characteristic (ROC) analysis was performed to determine the predictive value of the HTT. RESULTS: Observed HTTs for malignant focal liver lesions (mean, 6.2 seconds; range, 2-10 seconds) were significantly lower than those for nonmalignant lesions (mean, 9.5 seconds; range, 4-25 seconds; P < .001). ROC analysis revealed cutoff values of 7 seconds for HTT and 0.879 for area under the ROC curve. For HTTs of 7 seconds or shorter, hepatic malignancies were detected with a sensitivity of 79%, a specificity of 80%, a positive predictive value of 53%, and a negative predictive value of 93%. No malignant lesions had an HTT longer than 10 seconds. CONCLUSION: HTT alone could be a good predictor for nonmalignancy of focal liver lesions.

Quantitative assessment of the transport of elastic and rigid vesicle components and a model drug from these vesicle formulations into human skin in vivo.

The aim of this study was to quantitatively assess the distribution profiles of elastic and rigid vesicle material in human skin in vivo. Furthermore, the distribution profiles of the model drug ketorolac applied in these vesicle formulations was investigated. A deuterium-labelled phospholipid was incorporated into these vesicles to serve as a marker for the vesicle material. The vesicles were loaded with ketorolac at saturated concentrations. Vesicle solutions were applied non-occlusively onto the skin and the treated site was sequentially tape-stripped. Tape-strips were analyzed for vesicle material using attenuated total reflectance-Fourier transform infrared spectroscopy and for ketorolac by extraction of the tape-strips followed by high pressure liquid chromatography. Distribution profiles in the stratum corneum (SC) were obtained for the elastic and rigid vesicle material and for the ketorolac. These profiles have suggested that elastic vesicle material can rapidly enter the deeper layers of the SC and can reach almost the SC-viable epidermal junction. Rigid vesicle material, however, did not penetrate deep into the SC. Furthermore, the elastic vesicles were better than the rigid vesicles in the enhancement of ketorolac transport into human SC. The distribution profile of ketorolac in the deeper SC layers was, however, different from that of the vesicle material. This suggests that once the elastic vesicles partition into the SC, the ketorolac is released from the vesicles. The elastic vesicles are superior to the rigid vesicles both in terms of vesicular transport into the SC and in terms of therapeutic potential as a skin delivery vehicle.

Phosphatidylazidothymidine and phosphatidyl-ddC: assessment of uptake in mouse lymphoid tissues and antiviral activities in human immunodeficiency virus-infected cells and in Rauscher leukemia virus-infected mice.

During the early stages of human immunodeficiency virus (HIV) infection, although symptoms are absent and viral replication in peripheral blood mononuclear cells is low, substantial levels of HIV replication can be documented in lymphoid tissue [G. Pantaleo, C. Graziosi, J.F. Demarest, L. Butini, M. Montroni, C.H. Fox, J.M. Orenstein, D.P. Kotler, and A.S. Fauci, Nature (London) 362:355-358, 1993, and J. Embretsen, M. Zupancic, J.L. Ribas, A. Burke, P. Racz, K. Tenner-Tacz, and A.T. Haase, Nature (London) 362:359-362, 1993]. This observation suggests that earlier treatment of HIV infection may be indicated and that strategies for enhancing drug targeting to the lymphoid tissue reservoris of HIV infection may be beneficial. To address this issue, we synthesized dioleoylphosphatidyl-ddC (DOP-ddC) and dipalmitoylphosphatidyl-3'-azido-3'-deoxythymidine (DPP-AZT), phospholipid prodrugs which form lipid bilayers and which are readily incorporated into liposomes. The anti-HIV activity of DOP-ddC was similar to that of ddC in HIV type 1-infected HT4-6C cells, but DPP-AZT was considerably less active than AZT in HT4-6C cells. Liposomes containing DOP-[3H]ddC or DPP-[3H]AZT administered intraperitoneally to mice produced greater levels of total radioactivity over time in plasma, spleen, and lymphoid tissue relative to the results with [3H]ddC and [3H]AZT, respectively. DPP-AZT administered intraperitoneally in liposomes as a single daily dose to mice infected with Rauscher leukemia virus prevented increased spleen weight and reverse transcriptase levels in serum with a dose-response roughly comparable to that of AZT given continuously in the drinking water. DOP-ddC, DPP-AZT, and lipid conjugates of other antiretroviral nucleosides may provide higher levels of drug over time in plasma and in lymph nodes and spleen, important reservoirs of HIV infection, and may represent an interesting alternative approach to antiviral nucleoside treatment of AIDS.