Cost-Effectiveness and Budget Impact Analysis of Apixaban and Rivaroxaban Versus Warfarin in the Prevention of Stroke in Patients With Non-Valvular Atrial Fibrillation (NVAF) in Iran.

INTRODUCTION: This study evaluates the cost-effectiveness of Apixaban and Rivaroxaban, compared to Warfarin, for stroke prevention in patients with non-valvular atrial fibrillation in Iran. METHOD: A Markov model with a 30-year time horizon was employed to simulate and assess different treatment strategies' cost-effectiveness. The study population comprised Iranian adults with NVAF, identified through specialist consultations, hospital visits, and archival record reviews. Direct medical costs, direct nonmedical, and indirect costs were included. Quality-adjusted life years (QALY) were assessed using an EQ-5D questionnaire. This study utilized a cost-effectiveness threshold of $11 134 per QALY. RESULTS: Apixaban demonstrated superior cost-effectiveness compared to Rivaroxaban and Warfarin. Over 30 years, total costs were lower in the Apixaban and Rivaroxaban groups compared to the Warfarin group ($126.18 and $109.99 vs. $150.49). However, Apixaban showed higher total QALYs gained compared to others (0.134 vs. 0.133 and 0.116). The incremental cost-effectiveness ratio for comparing Apixaban to Warfarin was calculated at -1332.83 cost per QALY, below the threshold of $11 134, indicating Apixaban's cost-effectiveness. Sensitivity analyses confirmed the robustness of the findings, with ICER consistently remaining below the threshold. Over 5 years (2024-2028) of Apixaban usage, the incremental cost starts at USD 70 250 296 in the first year and gradually rises to USD 71 770 662 in the fifth year. DSA and PSA were assessed to prove the robustness of the results. CONCLUSION: This study shows that Apixaban is a cost-effective option for stroke prevention in non-valvular atrial fibrillation patients in Iran compared to Warfarin.

Effects of sesame dehulling on physicochemical and sensorial properties of its oil.

At present, sesame oil is extracted from un-hulled white sesame seeds by using cold press lubrication machines in local stores in Iran. This study aimed to evaluate the physicochemical properties and safety parameters of the hulled and un-hulled white sesame oils. The fatty acid composition, antioxidant activity, oxalates content, total phenolic content, carotenoid content, acid value, peroxide value, p-anisidine value, value total oxidation value (TOTOX), aflatoxins and pesticides residue, smoke point, color, relative density, and refractive index of oil sample were examined immediately after extracting the oil. The peroxide, p-anisidine, and TOTOX value of the hulled and un-hulled sesame oil samples were also examined periodically. After 7 months, the quality parameters were high and the oil samples were not consumable. Linoleic and oleic acids were the predominant fatty acids in the hulled and un-hulled sesame oils. The results of this study showed that the oil extracted from raw un-hulled sesame had a lower initial quality than hulled sesame oil and was oxidized more rapidly than it during the storage period. Virgin oils contained impurities acting like prooxidants and reduced their stability and shelf life. In addition, the un-hulled sesame oil contained higher amounts of antinutrient compounds (e.g., oxalate and pesticide residues) than the hulled sesame oil. Aflatoxin was not detected in our oil samples.

Cost-Effectiveness Comparison between Ticagrelor and Clopidogrel in Acute Coronary Syndrome in Iran.

Background: The present study aimed to determine the cost-effectiveness of ticagrelor compared with clopidogrel in Iranian patients with acute coronary syndrome (ACS). Methods: A 1-year decision tree model combined with a 20-year Markov transition model was used to simulate the long-term cost and effectiveness of both ticagrelor and clopidogrel in Iran based on an Iranian payer's perspective. Clinical efficacy data were extracted from the PLATO trial and other published studies. Costs were estimated based on local prices in public sectors. Deterministic and probabilistic sensitivity analyses were used to test the robustness of base-case results over the uncertainties of model inputs. All calculations, analyses, and modeling were done in TreeAge 2011 and Microsoft Excel 2013. Results: Compared with clopidogrel, the treatment of Iranian ACS patients with ticagrelor for 20 years resulted in an additional cost of US$ 2.39 in a hypothetical cohort of 1000 patients. However, ticagrelor led to 7.2 quality-adjusted life-years (QALYs) gained per 1000 hypothetical patients. Accordingly, the estimated incremental cost-effectiveness ratio for this analysis was US$ 332.032 per 1 QALY gained. Conclusion: Ticagrelor was a cost-effective antiplatelet medicine compared with clopidogrel in Iranian patients with ACS. This could help Iran's policymakers to allocate resources more efficiently to ACS.

Identification of Tartrazine adulteration and evaluating exposure to synthetic dyes of sunset yellow and Quinoline yellow through consumption of food products among children.

Excessive consumption of synthetic food dyes by children may raise concerns about their health. These dyes may aggravate the hyperactivity symptoms and exacerbate asthma in sensitive children. The purpose of this study was to determine the presence of sunset yellow and quinoline yellow dyes, as well as tartrazine in dairy-free fruit ice cream, freeze pop, jelly, and candy. Additionally, we evaluated the amount of two food dyes consumed by children. To do so, a total of 150 food samples, including 20 dairy-free fruit ice creams, 25 freeze pops, 57 jelly products, and 48 types of candy were randomly selected from stores in Shiraz, Iran. Then, using the high-performance liquid chromatography (HPLC) method and an ultraviolet (UV) detector, we measured the amounts of sunset yellow and quinoline yellow dyes and identified the use of tartrazine. Also, the per capita consumption (grams per day) of the mentioned foods was calculated using a checklist in two groups of male and female primary schoolchildren aged 6-9 years and 10-13 years in Shiraz, Iran. According to the results, 11 (7.33%) samples contained only tartrazine and 107 (71.33%) samples contained quinoline yellow and sunset yellow synthetic dyes. In addition, of 107 samples that used quinoline yellow and sunset yellow, 102 (95.33%) contained unauthorized tartrazine. Only seven (6.54%) samples contained exceedingly high concentrations of authorized quinoline yellow and sunset yellow synthetic dyes. However, the exposure assessment showed that the intake of quinoline yellow and sunset yellow was at average levels and the 95th percentile in both age groups was less than the associated acceptable daily intake (ADI). For synthetic dyes, the target hazard quotient (THQ) and hazard index (HI) were less than one, indicating that ingestion of these two dyes via food products does not pose a risk to children's overall health.

Development of fortified probiotic dairy desserts with added date extract, whey protein, inulin, folic acid, vitamin D and calcium.

This study aimed to develop fortified dairy desserts containing Lactobacillus casei and evaluate the physicochemical, sensory, and microbiological characteristics of the product during 28 days of storage. Seven dairy desserts were formulated by date extract (8%), whey protein (1.56%), inulin (4%(, folic acid (0.00066%), vitamin D (0.002%) and gluconate calcium (0.66%). The addition of date extract and inulin increased total solids while whey protein incorporation into dairy desserts led to the improvement of protein and phosphorous content. Furthermore, all fortified dairy desserts showed higher antioxidant activity and total phenolic content. Fortification of dairy desserts had no negative effect on the sensory acceptability and syneresis was not observed. In addition, the pH reduction and increased acidity did not adversely affect the count of L. casei, which remained above 8 log CFU g-1 during storage. Consequently, the fortified dairy dessert developed in this research is an innovative food product with good acceptability and high nutritional quality.

Short quality of life scale: A cross-cultural validation in Iranian patients with multiple sclerosis.

Health-related quality of life (HRQoL) is the prioritized measure in multiple sclerosis (MS) patients. The short quality of life scale (SQoL) developed by Devy et al. (2013) is an MS-specific and abbreviated scale with ten items suitable for routine medical care settings. The current study reported the cross-cultural validation of the scale in the Persian language. A total of 455 convenient MS patients with a mean age of 38.39 (9.28) ranged from 18 to 64 filled out the primary measure and the validating measures, including hospital anxiety and depression scale, visual analogue scale - quality of life, and a single index of the number of past-year MS relapse. The confirmatory factor analysis on original structure indicated an acceptable model fit. However, a modestly modified structure composing of physical-functional dimension (items #1-3), mental dimension (items #5-8), and pain & energy dimension (items 4 & 9,10) was also exposed with a sound fit and a meaningful structure. The overall internal consistency reliability was sound (0.88), and the concurrent validity was confirmed. The Persian short quality of life scale (P-SQoL) is the first translated and validated version of the scale, surfacing significant implications. Further cross-cultural investigations are recommended to re-examine current findings. The classic and recent suggestions concerning the close interplay between the immunity system and the psychological system and the implications based on Iran's context are discussed.

Effects of vitamin D-fortified oil intake versus vitamin D supplementation on vitamin D status and bone turnover factors: A double blind randomized clinical trial.

PURPOSE: The aim of this study was to compare the effect of vitamin D fortified oil consumption and vitamin D supplementation on serum 25-hydroxy vitamin D and bone turnover factors. METHODS: This study was a double-blind, parallel, randomized controlled clinical trial conducted over 12 weeks on 93 healthy participants aged 18-30 years. Participants were randomly allocated to 1 of the 3 groups: (a) supplement (a tablet of 1000 IU vitamin D supplement + 25 g canola oil daily, n = 31); (b) fortified oil (a placebo tablet + 25 g canola oil fortified with 1000 IU vitamin D daily, n = 30) and (c) control (a placebo tablet + 25 g canola oil, n = 32). Before and after the intervention 25-hydroxy vitamin D (25(OH)D1), parathyroid hormone (PTH2), bone alkaline phosphatase (BAP3) and collagen type 1 cross-linked C-telopeptide I (CTX4) were assessed. RESULTS: Serum 25(OH)D increased more in the vitamin D supplement group compared to the controls (P = 0.001). In addition, subgroup analysis revealed that just in the vitamin D sufficient subgroup, serum 25(OH)D increased more in both vitamin D supplement group and vitamin D fortified oil group, compared to the controls (P = 0.001 for both). The mean differences of PTH, BAP, and CTX were not significantly different among the study groups. CONCLUSION: Consumption of 1000 IU vitamin D per day for 12 weeks as a supplement or fortified oil could enhance the serum vitamin D in main population. However, the protective effect of vitamin D supplementation and oil fortification was seen just in vitamin D sufficient subgroup, not vitamin D deficient one. Besides, this dose of vitamin did not have a noticeable effect on bone turnover markers in this period. Registered under Iranian Registry of Clinical Trials (IRCT.ir) with ID number of: IRCT20180708040401N1.

Sphingomyelin liposome bearing whole Leishmania lysate antigens induce strong Th2 immune response in BALB/c mice.

OBJECTIVES: Whole Leishmania lysate antigens (WLL) has been shown to be effective to tackle leishmaniasis in murine models. Although liposomes can be considered as promising vaccines, the activity of phospholipase-A (PLA) in WLL, breeds difficulties to preparing stable liposomal WLL. One strategy to overcome this shortcoming is to use lipids such as sphingomyelin (SM) which is resistant against PLA. This study aim is formulating stable SM liposomes containing WLL and comparing their adjuvant effects with another first generation vaccine , i.e. solube Leishmania Antigen (SLA) liposomes in BALB/c mice. MATERIALS AND METHODS: BALB/c mice were immunized subcutaneously, three times with 2-week intervals, with Empty-liposome (E-lipo), Particulate WLL, Liposome-WLL, Liposome-SLA and control Buffer, three times every 2-week. Protection was assessed through measuring the swollen footpads and the load of parasites in the spleen. Other factors were used to assess the response of immune system by means of IgG subclasses, IL-4 and IFN-γ levels and intracellular cytokine assay in cultured splenocytes. RESULTS: Although liposomal WLL were stable in terms of physicochemical properties, mice received Liposome-WLL did not reduce footpad swelling. The load of parasites in spleen and levels of IL-4- were also higher compared to other immunized groups. In terms of IgG isotypes, no considerable difference observed in mice received Liposome-WLL or other formulations. CONCLUSION: Liposome-WLL could be a suitable vaccine delivery system when a Th2 response is desired. Also, further studies are warranted to fully understand the role of sphingomyelin in inducing an immune response.

Preparation of nanoliposomes containing HER2/neu (P5+435) peptide and evaluation of their immune responses and anti-tumoral effects as a prophylactic vaccine against breast cancer.

HER2/neu is an immunogenic protein inducing both humoral and cell-mediated immune responses. The antigen-specific cytotoxic T lymphocytes (CTLs) are the main effector immune cells in the anti-tumor immunity. To induce an effective CTL specific response against P5+435 single peptide derived from rat HER2/neu oncogene, we used a liposome delivery vehicle. In vivo enhancement of liposome stability and intracytoplasmic delivery of peptides are the main strategies which elevate the liposome-mediated drug delivery. Liposomes containing high transition temperature phospholipids, such as DSPC, are stable with prolonged in vivo circulation and more accessibility to the immune system. Incorporation of DOPE phospholipid results in the effective delivery of peptide into the cytoplasm via the endocytotic pathway. To this end, the P5+435 peptide was linked to Maleimide-PEG2000-DSPE and coupled on the surface of nanoliposomes containing DSPC: DSPG: Cholesterol with/without DOPE. We observed that mice vaccinated with Lip-DOPE-P5+435 formulation had the highest number of IFN-γ- producing CTLs with the highest cytotoxic activity that consequently led to significantly smallest tumor size and prolonged survival rate in the TUBO mice model. In conclusion, our study indicated that the liposomal form of P5+435 peptide containing DOPE can be regarded as a promising prophylactic anti-cancer vaccine to generate potent antigen-specific immunity.

A Phospholipase-A Activity in Soluble Leishmania Antigens Causes Instability of Liposomes.

AIM: This study aimed to investigate the existence of phospholipase-A (PLA) activity in Soluble L. major Antigens (SLA) because of no reports for it so far. Liposomes were used as sensors to evaluate PLA activity. OBJECTIVES: Liposomal SLA consisting of Egg Phosphatidylcholine (EPC) or Sphingomyelin (SM) were prepared by two different methods in different pH or temperatures and characterized by Dynamic Light Scattering (DLS) and Thin Layer Chromatography (TLC). METHODS: Lipid hydrolysis led to the disruption of EPC liposomal SLA in both methods but the Film Method (FM) produced more stable liposomes than the Detergent Removal Method (DRM). RESULT: The preparation of EPC liposomal SLA at pH 6 via FM protected liposomes from hydrolysis to some extent for a short time. EPC liposomes but not SM liposomes were disrupted in the presence of SLA. CONCLUSION: Therefore, a phospholipid without ester bond such as SM should be utilized in liposome formulations containing PLA as an encapsulating protein.

Development of a topical liposomal formulation of Amphotericin B for the treatment of cutaneous leishmaniasis.

BACKGROUND: Currently, there is no topical treatment available for any form of cutaneous leishmaniasis (CL) in most of the endemic areas. The aim of the current study was to develop a topical nano-liposomal Amphotericin B (AmB) for the treatment of CL. METHODOLOGY/PRINCIPAL FINDINGS: Liposomes containing 0.1, 0.2 and 0.4% AmB (Lip-AmB) were formulated and characterized for the size, entrapment efficiency, long term stability, and skin penetration properties using Franz diffusion cells. Liposomes diameters were around 100 nm with no change during more than 20 months' storage either at 4 °C or at room temperature. Franz diffusion cells studies showed that almost 4% of the applied formulations penetrated across the skin and the highest skin retention (73.92%) observed with Lip-AmB 0.4%. The median effective doses (ED50), the doses of AmB required to kill 50% of L. major amastigotes were 0.151, 0.151, and 0.0856 (µg/mL) in Lip-AmB 0.1, 0.2, 0.4%, respectively. Lip-AmB 0.4% caused 80% reduction in fluorescence intensity of GFP+ L. tropica infected macrophages at 5 µg/mL of AmB concentration. Topical Lip-AmB was applied twice a day for 4 weeks to the skin of BALB/c mice to treat lesions caused by L. major. Results showed the superiority of Lip-AmB 0.4% compared to Lip-AmB 0.2 and 0.1%. The parasite was completely cleared from the skin site of infection and spleens at week 8 and 12 post-infection in mice treated with Lip-AmB 0.4%. The results suggest that topical Lip-AmB 0.4% may be a useful tool in the treatment of CL and merits further investigation.

Thermal Stability and Kinetic Study on Thermal Degradation of Vitamin D3 in Fortified Canola Oil.

Nowadays, fortified vegetable oils with vitamin D3 are widely available in different countries and are consumed daily. The reduction rate of added vitamin D3 in fortified canola oil during heating process, the changes in oxidative status, and the thermal kinetic degradation of vitamin D3 in the fortified oil were investigated. For this purpose, canola oil was fortified at two levels of vitamin D3 with 5.625 µg/mL (low concentration or LC) and 13.585 µg/mL (a high concentration or HC). Samples were heated isothermally at 100, 150, and 180 °C for 30 min. The vitamin D3 concentration was determined by the high-performance liquid chromatographic method. The retention of vitamin D3 in samples treated at 100 °C for 30 min showed no significant reduction. Samples treated at 150 and 180 °C depending on the initial concentration showed the retention of 67.5% to 72.97% and 33.16% to 40.35% of vitamin D3 , respectively. An inverse relationship was found between the increment of lipid oxidation products (peroxide and anisidine values) and the retention of vitamin D3 . Kinetic parameters such as rate constant, activation energy, decimal reduction time, and quotient indicator were also calculated. An Arrhenius relationship was used for the assessment of temperature dependence of vitamin D3 degradation. Activation energies for vitamin D3 in LC and HC between 100 and 180 °C were found to be 44.01 and 38.77 kJ/mol, respectively. PRACTICAL APPLICATION: The oil can be fortified with vitamin D3 at low cost and offers a good bioavailability. A high-temperature cooking method may not be appropriate for the fortified products containing high lipid content.

Interactive anticancer effect of nanomicellar curcumin and galbanic acid combination therapy with some common chemotherapeutics in colon carcinoma cells.

OBJECTIVE: In the current investigation, we aimed to study the combined cytotoxicity of curcumin, as a nanomicellar formulation, and galbanic acid (Gal), dissolved in DMSO against the murine C26 and human Caco-2 colon carcinoma cells. Further, curcumin potential for cisplatin and doxorubicin (Dox) co-therapy was studied. MATERIALS AND METHODS: The combined cytotoxic effect of these phytochemicals at varying dose ratios were examined using the MTT colorimetric assay. Moreover, the time-dependent toxicity of curcumin, cisplatin, Dox, and pegylated liposomal Dox (Doxil) was determined. The interactive anti-proliferative behavior of these compounds was examined using the CompuSyn software. RESULTS: Nanomicellar curcumin showed considerable cytotoxicity in C26 cells 24 hr post-treatment. Co-treatment of cells with curcumin nanomicelles: Gal had a synergistic effect in C26 (at 10:1 molar ratio), and Caco-2 (at 1:5 molar ratio) cell lines in cell cultures. Nanomicellar curcumin showed strong and mild synergistic inhibitory effects in C26 cells when co-administered with Doxil and cisplatin, respectively. CONCLUSION: Curcumin nanomicelles and Gal had a synergistic effect in C26 and Caco-2 cell lines. It is speculated that nanomicellar curcumin shows synergistic cancer cell killing if administered 24-hr post-injection of Doxil and cisplatin.

P435 HER2/neu-derived peptide conjugated to liposomes containing DOPE as an effective prophylactic vaccine formulation for breast cancer.

The present study was aimed to develop an effective nanoliposomal vaccine delivery system with P435 HER2/neu-derived peptide conjugated to Maleimide-PEG2000-DSPE. The nanoliposome formulation composed of DSPC/DSPG/Chol/DOPE and monophosphoryl lipid A was used as an adjuvant. Liposomal formulations were prepared and their physical properties were characterized. Anti-tumoral efficacy of formulations was evaluated by immunization of tumor-bearing BALB/c mice and the generated immune response was studied by using ELISpot and flow cytometry analysis. The results of the study demonstrated Lip + DOPE + P535 formulation caused the lowest tumor size and the longest survival time in TUBO mice model and could make it a promising candidate in developing effective vaccines against HER2-positive breast cancers.

Preparation of nanoliposomes linked to HER2/neu-derived (P5) peptide containing MPL adjuvant as vaccine against breast cancer.

The study was aimed at evaluating antitumor and immunomodulatory effects of liposomal vaccine composed of P5 human epidermal growth factor receptor 2 (HER2)/neu-derived peptide coupled to the surface of high-temperature nanoliposomes containing distearoylphosphocholine:distearoylphosphoglycerol:Chol:dioleoylphosphatidylethanolamine (DOPE) comprising monophosphoryl lipid A (MPL) adjuvant in HER2/neu overexpressing the breast cancer model. BALB/c mice bearing TUBO carcinoma were subcutaneously immunized with formulations containing 10 µg P5 peptide and 25 µg MPL three times with 2-week intervals. To determine immuno responses in immunized mice, the amount of released interferon-γ and IL-4 were measured by the enzyme-linked immunospot method and the flow cytometric analysis on the isolated splenocytes. The results demonstrated that tumor-bearing mice immunized with Lip/DOPE/MPL/P5 formulation had the most released interferon-γ and the highest cytotoxic T lymphocyte responses that led to the lowest tumor size and the longest survival time than those of other formulations. The results achieved by Lip/DOPE/MPL/P5 formulation could make it a suitable candidate to induce effective antigen-specific tumor immunity against breast cancer.

Cationic liposomes formulated with a novel whole Leishmania lysate (WLL) as a vaccine for leishmaniasis in murine model.

Although there have been numerous attempts to develop a successful vaccine against leishmaniasis, based on the clinical trial in this field, no vaccine against Leishmania in routine way can be found for globally effective vaccination in human. Amongst, first generation vaccines consisting of parasite fractions or whole killed Leishmania showed more successful results in clinical trials. It seems that the main reason for the low efficacy of these vaccines is lack of a suitable adjuvant. In this study, a crude extract of detergent-solubilized L. major promastigotes as a novel developed antigen (whole Leishmania lysate (WLL)) was formulated in liposomal form. The cationic liposomes consisting of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) were used to deliver WLL. Liposomes formulations containing different WLL concentrations (prepared from 103, 104, 105, 106 and 107 parasites) were prepared and characterized for particle size, surface charge, proteins, DNA and phospholipids contents. Moreover, to explore the type of immune response generated and extend of immunization, in vivo and in vitro tests including evaluation of lesion development, parasite burden in the foot and spleen, Th1 and Th2 cytokine analysis, and titration of IgG isotypes before and after the challenge were used. The maximum immunization was provided by WLL06 as depicted by the reduction of footpad swelling andparasite load, increase in anti-Leishmania IgG2a production, though no significant difference was observed between mice which received WLL05 vs WLL06. While maximum immunization was seen in WLL06 group, most of the liposomal WLL formulations induced a mixed Th1/Th2 response. Hence, a more protective immune response is expected to be induced when an immune potentiator adjuvant such as CpG ODNs would be co-deliverd in WLL liposomal formulations.

Stimulation of Tumor-Specific Immunity by p5 HER-2/neu Generated Peptide Encapsulated in Nano-liposomes with High Phase Transition Temperature Phospholipids.

BACKGROUND: The aim of this study is to evaluate the possible advantages of liposomes with high transition temperature (Tm) in the function of a vaccine for P5 HER2/neu-generated peptide and its adjuvant action to elicit CD8+ T cell response and its efficacy in TUBO in vivo tumor mice model, which over expresses the HER2/neu oncogene. P5, a hydrophobic peptide, was encapsulated in the nanoliposomes consisting of DSPC/DSPG/Chol(Tm 54°C) with a chaotropic loading system via 7M urea and described by size, zeta potential, encapsulation efficiency, and the structural stability of SDS-PAGE. METHODS: We immunized the mice for three times subcutaneously based on a two-week intervals using encapsulated peptide in the nanoliposomes, empty liposome, P5 in PBS, and PBS. Enzyme-linked immunospot assay, cytotoxicity test, and flow cytometric studies followed by the size of tumor and survival time measurements, which were done in TUBO tumor mice version. RESULTS: Findings of ELISpot and flow cytometric analysis showed that immunization with Lip-p5 nanoliposomes has enhanced the antigen-specific IFN-γ response of CD8+ T cells and induced CTL response, which resulted in a smaller tumor and longer survival time. In addition to increase in amounts of IFN-γ-CD8+ T cells in a group, which was immunized with Lip-P5, our findings also revealed a Th1 shift in the group immunized with an empty liposome with reduced frequencies of IL-4-producing cells and increase of IFN-γ-producing cells. CONCLUSION: The results indicated that simple liposomes consisting of phospholipids with high transition temperature could be an effective vaccine vehicle for tumor-associated antigens for inducing cell mediate immunity.

Cationic Immune Stimulating Complexes Containing Soluble Leishmania Antigens: Preparation, Characterization and in Vivo Immune Response Evaluation.

BACKGROUND: Cationic immune stimulating complexes (PLUSCOMs) are particulate antigen delivery systems. PLUSCOMs consist of cationic immunostimulatory complexes (ISCOMs) derivatives and are able to elicit in vivo T cell responses against an antigen. OBJECTIVE: To evaluate the effects of PLUSCOMs containing Leishmania major antigens (SLA) on the type of immune response generated in the murine model of leishmaniasis. METHODS: PLUSCOMs consisting of 1, 2-dioleoyl-3-trimethylammonium-propane (DOTAP) were used as antigen delivery system/immunoadjuvants for soluble SLA. BALB/c mice were immunized subcutaneously, three times in 2-week intervals. Footpads swellings at the site of challenge and parasite loads were assessed as a measure of protection. The immune responses were also evaluated by determination of IgG subclasses and the level of IFN-γ and IL-4 in cultured splenocytes. RESULTS: There was no significant difference (p<0.05) between the sizes of lesions in mice immunized with different formulations. Also, there was no significant difference in the number of parasites in the footpad or spleen of all groups compared with the control group. The highest level of IFN-γ secretion was observed in the splenocytes of mice immunized with PLUSCOM/SLA (p<0.001) and lower amounts of IL-4 was observed in PLUSCOM group (p<0.001) as compared to negative control. CONCLUSION: Our results indicated that SLA in different formulations generated an immune response with mixed Th1/Th2 response that was not protective enough despite the activation of CD4+ T cells with secreting IFN-γ in groups which received PLUSCOM with antigen.

Optimizing the therapeutic efficacy of cisplatin PEGylated liposomes via incorporation of different DPPG ratios: In vitro and in vivo studies.

The anionic lipid DPPG is known to enhance the cellular uptake of liposomes by forming phase boundaries of high fusogenic potentials in vesicular membranes. The focus of this study is to optimize DPPG concentrations to improve the therapeutic efficacy of cisplatin-loaded liposomes. First, cisplatin liposomes composed of HSPC, mPEG2000-DSPE and cholesterol with increasing amounts of DPPG (10, 20 and 30% mol) were prepared by ethanol injection. Liposomes were then characterized by their size, zeta potential and cytotoxicity against C26 colon carcinoma cells. In an experimental system, based upon C26 tumor bearing BALB/c, mice were treated with administering i.v. doses of different formulations, once weekly for total of three weeks. Although with the highest DPPG ratio (30% mol) liposomes exhibited the highest toxicity in vitro, at 10% DPPG better stability of the encapsulated drug was obtained in the presence of serum. In addition, survival of animals was substantially improved at 10% DPPG compared to the higher DPPG contents. It is thus presumable that the high density of negatively charged residues of DPPG gave rise to repulsive forces between phospholipids in concentric lipid bilayers, which resulted in the instability of lipid structure and the subsequent premature drug leakage. Results indicated that cisplatin liposome fabricated with the inclusion of 10% DPPG, maintains the stability while in circulation, and improves therapeutic efficacy due to fusogenic properties; therefore might serve as an effective and stable formulation of cisplatin. However, further investigations are required to confirm the potential anti-tumor effects of cisplatin anionic nanoliposomes in various tumor types.

Immunoliposomes containing Soluble Leishmania Antigens (SLA) as a novel antigen delivery system in murine model of leishmaniasis.

Development of new generation of vaccines against leishmaniasis requires adjuvants to elicit the type and intensity of immune response needed for protection. The coupling of target-specific antibodies to the liposomal surface to create immunoliposomes has appeared as a promising way in achieving a liposome active targeting. In this study, immunoliposomes were prepared by grafting non-immune mouse IgG onto the liposomal surface. The influence of active targeted immunoliposomes on the type and intensity of generated immune response against Leishmania was then investigated and compared with that of liposomes and control groups which received either SLA or HEPES buffer alone. All formulations contained SLA and were used to immunize the mice in the left hind footpad three times in 3-week intervals. Evaluation of lesion development and parasite burden in the foot and spleen after challenge with Leishmania major, evaluation of Th1 cytokine (IFN-γ), and titration of IgG isotypes were carried out to assess the type of generated immune response and the extent of protection. The results indicated that liposomes might be effective adjuvant systems to induce protection against L. major challenge in BALB/c mice, but stronger cell mediated immune responses were induced when immunoliposomes were utilized. Thus, immune modulation using immunoliposomes might be a practical approach to improve the immunization against L. major.