The objective of this study was to optimize the in vivo activity of proteolipid protein (PLP)-bifunctional peptide inhibitor (BPI) molecule to suppress experimental autoimmune encephalomyelitis (EAE) in SJL/J mice and evaluate pharmacokinetic profiles of PLP-BPI. PLP-BPI is constructed via conjugation of myelin PLP 139-151 with CD11a 237-246 -derived peptide (LABL) via a spacer. The hypothesis is that PLP-BPI binds simultaneously to major histocompatibility complex-II and intercellular adhesion molecule-1 on the antigen-presenting cell (APC) and inhibits the formation of the immunological synapse during T-cell and APC interactions. In this study, the structure of BPI was modified by varying the spacer and was evaluated in the EAE model.Intravenous injections of BPI derivatives inhibited the onset, severity, and incidence of EAE more effectively and induced a lower incidence of anaphylaxis than that produced by unmodified PLP-BPI. As anticipated, production of interleukin-17, a proinflammatory cytokine commonly found in elevated levels among multiple sclerosis (MS) patients, was significantly lower in Ac-PLP-BPI-PEG6-or Ac-PLP-BPI-NH 2 -2-treated mice than in phosphate-buffered saline-treated mice. These results suggest that BPI-type molecules can be modified to achieve more efficient and better tolerated BPI-based derivatives for the treatment of MS.

Abstract.A study on thalassemia intermedia and major patients in Jakarta was initiated to obtain a comprehensive picture of metabolic dysregulation, iron overload, oxidative stress, and cell damage. Data are presented from a group of 14 transfusiondependent patients in an age range of 11-25 years (T) and another group of 9 frequently transfused (for at least 15 years) patients aged 17-30 years (L). A third group comprised 6 patients (aged 7 to 14 years) who had not yet obtained transfusions (N). The 21 controls (C) were voluntary students without diagnosis or clinical signs of thalassemia up to 30 years of age. The study was approved by the Ethical Clearance Board of the Medical Faculty and all blood samples from controls and patients were obtained on fully informed consent. Levels of antioxidants (vitamins A, C, E and β-carotene) and reactive thiols are considerably decreased in transfused patients, whereas signs of iron overload and cell damage are increased (serum iron, ferritin, transferrin saturation, SGOT, SGPT, γ-GT, bilirubin). Results can be summarized that non-transfused thalassemia intermedia patients exert slight signs of oxidative stress, and increased hemoglobin degradation but no significant indication of tissue or cell damage. This picture differs considerably from transfusion-dependent thalassemia major patients: highly significant decrease in antioxidants and thiols and tremendous iron overload and cell damage. The picture is even worsened in long-term transfused patients. Iron chelation after transfusion is not sufficient in Indonesia, because it is normally (with few exceptions) applied only once together with transfusion. Hence, one major reason of the bad condition of transfusion-dependent thalassemia patients in Indonesia appears to be frequent transfusions (on the average one per month) and insufficient chelation of one treatment per month together with transfusion.

Erythrocyte membrane structural parameters were studied in transfusion-dependent beta-thalassemia patients, in long-term transfused patients (regularly transfused < 15 years), and in those who had not yet obtained transfusions. Controls were voluntary students up to 30 years of age without diagnosis or clinical signs of thalassemia. Membranes were isolated and investigated by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electron paramagnetic resonance (EPR) spectroscopy. Data obtained from the thiol-reactive spin label N-ethyl-maleimidoproxyl reveal immobilization of protein environment in erythrocyte membranes from thalassemic patients. SDS-PAGE shows both degradation and aggregation of membrane proteins. Thalassemic erythrocyte membranes exert higher order parameters in the hydrophobic region as determined by 16-doxyl-stearic acid. Rotational correlation times of this spin label increase only in transfused patients. Polarity is higher in membranes of all patients than in controls. In the polar interface, order parameters obtained from 5-doxyl-stearic acid increase in non-transfused and decrease in transfusion-dependent patients as compared with controls. Transfused patients exert increasing membrane order in the hydrophobic region and counter-currently decreasing order in the polar interface indicating loss of membrane integrity along with the loss of fluidity and polarity gradients and the loss the energetic barrier function of the membrane.

Latar belakang: Kami melakukan penelitian selama 10 tahun terakhir untuk mendapatkan gambaran yang komprehensif mengenai kelebihan besi, stres oksidatif, dan kerusakan sel pada pasien talasemia di Jakarta. Metode: Dari sampel darah 15 pasien yang bergantung transfusi (kelompok T), 5 pasien yang belum ditransfusi (kelompok N) dan 10 kontrol (kelompok C) dilakukan analisis plasma dan membran eritrosit yang diisolasi dan strukturnya diteliti dengan spekroskopi electron paramagnetic resonance (EPR) menggunakan doxyl-stearic acid dan maleimido-proxyl spin lable. Data statistik dianalisis dengan ANOVA. Hasil: Terdapat nilai trigliserida lebih tinggi dan nilai kolesterol plasma lebih rendah pada penderita talasemia dibandingkan dengan kontrol. Kadar vitamin E (kelompok C: 21.8 vs T: 6.2 μmol/L) dan reactive thiols (C: 144 vs. T: 61 μmol/L) jauh lebih rendah pada pasien yang bergantung-transfusi. Tanda-tanda stres oksidatif (MDA, C: 1.96 vs. T: 9.2 μmol/L) dan kadar transaminase dalam serum sangat tinggi. Pasien belum ditransfusi memperlihatkan sedikit tanda-tanda terjadinya stres oksidatif, tetapi tidak terdapat tanda-tanda yang berarti adanya kerusakan jaringan atau sel. Parameter membran sel darah merah dari spektroskopi electron paramagnetic resonannce (EPR) menunjukkan hasil yang sangat berbeda-beda untuk setiap kelompok. Pada pasien bergantung-transfusi, struktur membran sel darah merah serta gradien polaritas dan fl uiditas dalam domain lipid menunjukkan rusak; kapasitas mengikat protein tiol di dalam membran lebih rendah dan statis.

No abstract