Evaluation of Liposomal and Microbubbles Mediated Delivery of Doxorubicin in Two-Dimensional (2D) and Three-Dimensional (3D) Models for Breast Cancer.

OBJECTIVE: Liposomal cancer treatment strategies are useful in removing the side effects that were the main concern in recent years. In this study, we prepared microbubble (MBs) conjugated with DOX-loaded liposomes (DOX-loaded MBs) and investigated their effectiveness in in vitro breast cancer cells in two dimensions (2D) and three dimensions (3D). MATERIALS AND METHODS: With this aim, breast cancer cells with different features (4T1, MDA-MB231, MCF-7) were growth in 2D and 3D dimensions. The cytotoxic and cell death effects under different conditions, durations and doses were evaluated with WST-1, trypan-blue, colony counts. Apoptotic effects were investigated with flow cytometric Annexin-V-PI and immunohistochemical (Ki-67, caspase 3, 8, 9) methods. RESULTS: After free DOX and LipoDOX were applied, the proliferation index of three cell lines reduced. Intrinsic and extrinsic apoptotic pathways were activated in both 2D and 3D models. However, this effect was observed at lower levels in the 3D model due to the difficulty of diffusion of DOX into the spheroids. Additionally, the suitability of the 3D model for breast cancer cells was supported by formation of ductus-like structures and spheroids. Cell deaths were not observed significantly with the DOX-loaded microbubbles due to rising of MBs to the surface and not reaching spheroids held in matrigel of 3D model. CONCLUSION: DOX and LipoDOX showed anti-proliferative and apoptosis-inducing effects in breast cancer cells. However, these effects indicated variability depending on the cell lines and 2D or 3D model types.

A Way to Increase the Sensitivity and Specificity of the Hamilton Depression and Anxiety Scales.

Objective: The Hamilton Depression Rating Scale (HDRS-17) and the Hamilton Anxiety Rating Scale (HARS-14) have been acknowledged as gold standards in evaluating the severity of depression and anxiety. The specificity and sensitivity of these scales in predicting somatic complaints of depression and anxiety are issues in both clinical and research areas. The present study proposes a new model to enhance the sensitivity and specificity of HDRS-17 and HARS-14 for predicting symptoms of insomnia, inappetence, and loss of libido in psychiatric patients. Methods: This study included 1507 patients diagnosed withbipolar disorder, depression, panic disorder, obsessive-compulsive disorder, and generalized anxiety disorder. The HDRS-17 and the HARS-14 were utilized as predictive scales for the prediction of patients' sleep, appetite, and libido. The sensitivity and specificity were computed using the receiver operating characteristic (ROC). Logistic regression was performed to enhance the predictive values. The predictive value of the logistic regression model was not satisfactory, and a conversion table was therefore designed for each symptom-diagnosis subgroup. The new joint ROC model was then used to recalculate the sensitivity and specificity of the 2 scales for each symptom-diagnosis subgroup. The outcome is a prediction table, presented for use by clinicians. Results: It was observed that the new statistical model, the joint ROC, increased the sensitivity and specificity of the HDRS-17 and the HARS-14. Conclusion: : Based on the results of the evaluations with the HDRS and the HARS, the joint ROC method was developed to better predict the presence of symptoms.

Myocardial ischemic memory imaging with molecular echocardiography.

BACKGROUND: Diagnosing acute coronary syndrome in patients presenting with chest discomfort is a challenge. Because acute myocardial ischemia/reperfusion is associated with endothelial upregulation of leukocyte adhesion molecules, which persist even after ischemia has resolved, we hypothesized that microbubbles designed to adhere to endothelial selectins would permit echocardiographic identification of recently ischemic myocardium. METHODS AND RESULTS: Lipid microbubbles (diameter, 3.3+/-1.7 microm) were synthesized. The selectin ligand sialyl Lewis(x) was conjugated to the microbubble surface (MB(sLex)). Control bubbles (MB(CTL)) bore surface Lewis(x) or sialyl Lewis(c). Intravital microscopy of mouse cremaster muscle was performed after intravenous injection of MB(sLex) (n=11) or MB(CTL) (n=9) with or without prior intrascrotal tumor necrosis factor-alpha. There was greater adhesion of MB(sLex) to inflamed versus noninflamed endothelium (P = 0.0081). Rats (n=12) underwent 15 minutes of anterior descending coronary artery occlusion. After 30 minutes and 1 hour of reperfusion, high-mechanical-index nonlinear echocardiographic imaging was performed in which single frames were acquired at 3.5 and 4 minutes after intravenous injection of MB(sLex) or MB(CTL). Video intensity at 4 minutes was subtracted from that at 3.5 minutes to derive target-specific acoustic signal. MB(sLex) caused greater opacification in postischemic versus nonischemic myocardium at both time points (P < or = 0.002). Immunostaining confirmed endothelial P-selectin expression in the ischemic bed. CONCLUSIONS: Echocardiographic identification of recently ischemic myocardium is possible using ultrasound contrast agents targeted to selectins. This may offer a new approach to the more timely and precise diagnosis of acute coronary syndrome in patients presenting with chest pain of uncertain cardiac origin.

Phase behavior of DSPC/PEG40St mixtures at higher emulsifier contents.

Phase behaviors of 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC) and polyoxyethylene(40)stearate (PEG40St) were investigated with Langmuir monolayer isotherms and Brewster angle microscopy (BAM) imaging at DSPC/PEG40St molar ratios ranging from 9:1 to 5:5. Two plateaus were found in the Langmuir isotherms which were relatively shorter for the 9:1 mixture and extended significantly by increasing the PEG40St content, indicating that the PEG40St squeezed out whereas more emulsifier retained in the monolayer at higher PEG40St contents. A strong hysteresis was observed when the mixed monolayers were subjected to compression-expansion cycles. The degree of hysteresis for the first cycles also increased with increasing PEG40St content in the monolayer. Gray scale intensities in the Brewster angle microscopy images were determined for pure DSPC and pure PEG40St and a scale was established to better interpret the morphologies for the mixtures. Bud and vessels formed during the PEG40St squeezed out upon compression. Upon expansion, PEG40St and DSPC is reappeared on the monolayer. When considered BAM images together with the Langmuir isotherm, PEG40St molecules were found to be well distributed within the DSPC molecules at lower DSPC/PEG40St mole ratios and mostly phase separated at higher mole ratios. It was concluded that higher PEG40St content would be advantageous for the design of an efficient and cheaper ultrasound contrast agents.

Lack of Association between Pulse Steroid Therapy and Bone Mineral Density in Patients with Multiple Sclerosis.

Multiple sclerosis (MS) has been associated with reduced bone mineral density (BMD). The purpose of this study was to determine the possible factors affecting BMD in patients with MS. We included consecutive 155 patients with MS and 90 age- and sex-matched control subjects. Patients with MS exhibited significantly lower T-scores and Z-scores in the femoral neck and trochanter compared to the controls. Ninety-four (61%) patients had reduced bone mass in either the lumbar spine or the femoral neck; of these, 64 (41.3%) had osteopenia and 30 (19.4%) had osteoporosis. The main factors affecting BMD were disability, duration of MS, and smoking. There was a negative relationship between femoral BMD and EDSS and disease duration. No association with lumbar BMD was determined. There were no correlations between BMD at any anatomic region and cumulative corticosteroid dose. BMD is significantly lower in patients with MS than in healthy controls. Reduced BMD in MS is mainly associated with disability and duration of the disease. Short courses of high dose steroid therapy did not result in an obvious negative impact on BMD in the lumbar spine and femoral neck in patients with MS.

Effect of the systemic use of methotrexate on the oxidative stress and paraoxonase enzyme in psoriasis patients.

Previous studies have indicated that oxidative stress contributes in the efficacy and toxicity of methotrexate (MTX) treatment. The present study aims to investigate the systemic MTX treatments impact on the total oxidant and antioxidant status of the patients with psoriasis. A total of 26 psoriasis patients were included in the study. Serum total oxidant status (TOS), total antioxidant status (TAS), and serum paraoxonase enzyme (PON) levels were measurement of all patients, and Oxidative Stress Index (OSI) were calculated before and after 8 weeks of MTX therapy. Psoriasis Area Severity Index scores of the patients decreased significantly after MTX treatment. While the serum concentrations of total cholesterol, low density lipoprotein, and high density lipoprotein decreased significantly, the serum ALT levels of the patients increased significantly after MTX treatment. There was no statistically significant alteration in serum levels of PON, TAS, TOS, and OSI after the MTX therapy. The oxidative stress emerging with 8-week MTX treatment is not significantly increased in the patients. In parallel with the decreasing inflammation by MTX treatment in patients with psoriasis, a decrease in oxidative stress (OS) is also expected. However, the expected reduction in OS might have been precluded by MTX-induced OS, which resulted in no significant difference between pre- and post-treatment values of OS parameters in our study. There is a possibility that the 8-week results may change with longer treatment durations and higher cumulative doses.