Differential dependencies on [Ca2+] and temperature of the monolayer spontaneous curvatures of DOPE, DOPA and cardiolipin: effects of modulating the strength of the inter-headgroup repulsion.

Biomembranes assume nonlamellar structures in many cellular events, with the tendency of forming a nonlamellar structure quantified by the monolayer spontaneous curvature, C(0), and with many of these events involving the acts of Ca(2+). Despite this biologically important intimacy, how C(0) is affected by [Ca(2+)] is unknown. In this study, we use the X-ray diffraction technique and the reconstruction of electron density profiles to measure the C(0)s of a zwitterionic phospholipid, DOPE, and two anionic phospholipids, DOPA and 18 : 1 (9Z) cardiolipin, at temperatures from 20 °C to 40 °C and [Ca(2+)]s from 0 mM to 100 mM; these phospholipids are chosen to examine the contributions of the electric charge density per molecule. While showing a strong dependence on temperature, C(0,DOPE) is nearly independent of [Ca(2+)]. In contrast, C(0,DOPA) and C(0),cardiolipin are almost unresponsive to the temperature change but affected by the [Ca(2+)] variation; and C(0,DOPA) varies with [Ca(2+)] ∼1.5 times more strongly than C(0,cardiolipin), with the phase preferences of DOPA and cardiolipin shifting to the H(II) phase and remaining on the Lα phase, respectively, at [Ca(2+)] = 100 mM. From these observations, we reveal the effects of modulating the strength of the inter-headgroup repulsion and discuss the mechanisms underlying the phase behaviour and cellular functions of the investigated phospholipids. Most importantly, this study recognizes that the headgroup charge density is dominant in dictating the phase behaviour of the anionic phospholipids, and that the unique molecular characteristics of cardiolipin are critically needed both for maintaining the structural integrity of cardiolipin-rich biomembranes and for fulfilling the biological roles of the phospholipid.

Long-term follow-up of prostate cancer patients treated with vaccine and definitive radiation therapy.

BACKGROUND: Vaccine therapy in combination with radiation therapy may improve distant and/or local control in prostate cancer. We present long-term follow-up data on the secondary and exploratory endpoints of safety and biochemical failure, respectively, from patients with clinically localized prostate cancer treated definitively with a poxviral vector-based therapeutic vaccine combined with external beam radiation therapy (EBRT). METHODS: Thirty-six prostate cancer patients received definitive EBRT plus vaccine. A total of 18 patients were treated with adjuvant standard-dose interleukin-2 (S-IL-2) (4 MIU m(-2)) and 18 were treated with very low-dose IL-2 (M-IL-2) (0.6 MIU m(-2)). Seven patients were treated with EBRT alone. Twenty-six patients treated with EBRT plus vaccine returned for follow-up, and we reviewed the most recent labs and clinical notes of the remaining patients. RESULTS: Median follow-up for the S-IL-2, M-IL-2 and EBRT-alone groups was 98, 76 and 79 months, respectively. Actuarial 5-year PSA failure-free probability was 78%, 82% and 86% (P=0.58 overall), respectively. There were no significant differences between the actuarial overall survival and the prostate cancer-specific survival between the two vaccine arms. Of the 26 patients who returned for follow-up, Radiation Therapy Oncology Group grade ≥2 genitourinary (GU) and gastrointestinal (GI) toxicity was seen in 19% and 8%, respectively, with no difference between the arms (P=1.00 and P=0.48 for grade ≥2 GU and GI toxicity, respectively). In all, 12 patients were evaluated for PSA-specific immune responses, and 1 demonstrated a response 66 months post-enrollment. CONCLUSIONS: We demonstrate that vaccine combined with EBRT does not appear to have significant differences with regard to PSA control or late-term toxicity compared with standard treatment. We also found limited evidence of long-term immune response following vaccine therapy.