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Cutaneous leishmaniasis is known as the most prevalent clinical form of leishmaniasis. It needs the development of new therapies due to the serious side-effects promoted by taking the current drugs. In the present study, dextran–behenic acid (DEX–BA) based nanomicelles were developed to direct the delivery of itraconazole (ITZ) to the macrophages and enhance its toxic effects against Leishmania parasites. DEX–BA was synthesized through the esterification of dextran with behenic acid. The critical micelle concentration of the newly developed conjugate was evaluated using pyrene as the fluorescent probe. The nanomicelles were generated by the dialysis method; then they were optimized by applying a Box–Behnken design. The effects of the dialysis temperature, polymer content, and sonication time on the characteristics of micelles were subsequently studied. Furthermore, in vitro efficacy against Leishmania major promastigotes and parasite-infected macrophages was evaluated. The optimized formulation showed the particle size of 195.16 ± 3.06 nm, the polydispersity index of 0.39 ± 0.01, the zeta potential of −16.29 ± 0.89 mV, the encapsulation efficiency % of 56.11 ± 4.9, and the release efficiency % of 51.29 ± 1.97. According to scanning electron microscopy, the nanomicelles were found to be nearly spherical in shape. ITZ-loaded nanomicelles showed the strongest anti-leishmanial activities when compared with the free ITZ and drug-free nanomicelles. It could be, therefore, concluded that ITZ-loaded nanomicelles might be useful as an alternative therapy for the treatment of cutaneous leishmania. [ABSTRACT FROM AUTHOR]

Leishmania infantum is an agent of visceral leishmaniasis (VL). Amastigote form is a more appropriate target for investigations on vaccines, treatment, and diagnosis. This study aimed to achieve the amastigotes of L. infantum in the golden hamster and J774 macrophages and report the pathological changes that occur in the liver and spleen of the hamsters with VL. 4 male golden hamsters were infected with L. infantum promastigotes. After 5 months, the hamsters were euthanized and touch and pathology smears were prepared from the livers and spleens. Then, these tissues were homogenized and centrifuged at 100×g. Supernatants were collected and centrifuged at 2000×g and the pellets were collected. In the next part of our study, J774 macrophages were infected with L. infantum promastigotes. Then, the infected macrophages were ruptured. Centrifuge stages were done same the previous part. The amastigotes were observed in touch and pathology smears. A load of amastigotes in the livers was more than the spleens in both types of smears. Although the livers’ structure had undergone pathological changes, the spleens were unchanged. Also, the macrophage infectivity ratio was up to 95%. Our results present a simple and accessible way of achieving a lot of pure and real amastigotes for different fields in Leishmania. Also, it seems that the pathological changes occurring in the spleen and the liver of animals with VL are different and probably can be attributed to the genetic and immune process of the infected animals. [ABSTRACT FROM AUTHOR]

Abstract: To investigate the effect of apolipoprotein B (apoB) on cell viability, we used lipid-free apoB as a model for denatured apoB. Lipid-free apoB had cytotoxicity to J774 macrophages, CHO cells and HepG2 cells, whereas apoB bound to low density lipoprotein (LDL) and lipid-free apolipoprotein A-I had no effect on cell viability. Lipid-free apoB induced apoptosis in J774 macrophages assessed by caspase-3 activation and annexin V binding. LDL receptor, heparan sulfate proteoglycans, and class A scavenger receptor were involved in the binding/uptake of lipid-free apoB, but lipid-free apoB binding/uptake by the cells did not correlate with cytotoxicity. Lipid-free apoB disrupted the lipid bilayer of large unilamellar vesicles containing calcein. We evaluated the interaction between apoB and cellular membrane by monitoring the change in intracellular Ca2+ concentration using Fura-2, and found that lipid-free apoB rapidly disrupted the cellular membrane in the absence or presence of the inhibitors for cellular binding/uptake mediated by the receptors. Therefore, it is suggested that lipid-free apoB induces cell death by disturbance of the plasma membrane. In addition to other lipid component in modified LDL, apoB itself has an ability to induce apoptosis and plays a crucial role in the development of atherosclerotic lesions. [Copyright &y& Elsevier]

In this study, we examined the signal transduction of dibutyryl cyclic adenosine monophosphate (dBcAMP) to stimulate the release of nitric oxide (NO) and interleukin-6 (IL-6) from J774 macrophages. These actions of dBcAMP were diminished by the presence of the inhibitors of protein kinase A (PKA), protein kinase C (PKC), p38 MAPK and nuclear factor-kappa B (NF-κB). In contrast, Go 6976 and PD98059 had no significant effects. Consistently, dBcAMP caused membrane translocation of PKCβII, δ, μ, λ and ζ isoforms, and increased atypical protein kinase C (aPKC) and p38 MAPK activities. The nuclear translocation and DNA-binding study revealed that dBcAMP stimulated NF-κB, activator protein-1 (AP-1), and CAAT/enhancer-binding protein (c/EBPβ). Via PKA, PKC and p38 MAPK-dependent signals, dBcAMP also induced inhibitory subunit of NF-κB (IκB) degradation, IκB kinase (IKK) activation, nuclear translocation of NF-κB subunit p65 and its association with the CREB-binding protein (CBP). These results illustrate that PKA activation in macrophages is able to stimulate PKC and p38 MAPK, which lead to IKK-dependent NF-κB activation and contribute to the induction of inducible nitric oxide synthase (iNOS) and IL-6 genes. [Copyright &y& Elsevier]