Background and Aim: It is a common practice for biomedical researchers to evaluate Leishmania infection indexes on in vitro cell cultures to address biological questions or test the efficacy of novel anti- parasitic compounds. In Leishmania infections that parasitize macrophages, infection indexes are usually determined by visual inspection of the cells directly using Giemsa staining. However, the mechanism of action of treatment on Leishmania parasites in this method is not evaluated, therefore motivating the finding of analysis approaches that allow simultaneously quantifying Leishmania infection index and cell death on in vitro cell cultures.

Methods: We proposed a method for simultaneously evaluating parasite infection indexes and cell death indexes using fluorescent microscopy. To perform such an analysis, we used a fluorescent dye mix of ethidium bromide and acridine orange (EB/AO), which binds nucleic acids.

Results: EB/AO achieved simultaneous quantification of infection index and death index under normal growth and treatment conditions that were comparable to the conventional Giemsa staining method while overcoming the drawbacks of the current method.

Conclusion: The cell death index and Leishmania infection index can be simultaneously quantified using our method, which is less user- skill- dependent and faster than the general test.

Background: Nanoliposomes are spherical nano-sized capsules enclosed by lipid membranes, serving as a biocompatible vehicle to enhance the delivery of therapeutic agents. The objective of this research is to prepare and characterize nanoliposome-encapsulated auraptene and compare its cytotoxic and anti-angiogenic effects to non-liposomal auraptene.

Methods: Liposomal auraptene was formulated using DSPC/ DSPG/ Cholesterol (molar ratio of 4:1:2) in combination with two different molar ratios of auraptene (0.1 and 0.05). The entrapment efficiency was evaluated using High- Performance Liquid Chromatography (HPLC). Various parameters, including Dynamic Light Scattering (DLS), zeta potential, stability, and release kinetics, were investigated. Subsequently, both liposomal and non-liposomal auraptene, along with bare liposomes, were applied to the MDA-MB-231 cell line for duration of 72 hours at 37°C at varying concentrations. Cytotoxicity was assessed using the MTT assay. Additionally, the study examined the anti-angiogenic effects on the vessels in the chorioallantoic membrane of chick embryos.

Results: The entrapment efficiency of auraptene was found to be satisfactory at 50%. The liposome size ranged from 85 to 241 nm, with a Z-Average of 190.9 nm. The zeta potentials for all formulations were consistently around -55.7, and the Polydispersity Index (PDI) was less than 0.3 for all formulations. The release profile demonstrated approximately 80% drug release over a period of 130 hours. Notably, liposomal auraptene exhibited a significantly lower IC₅₀ value (38.61 (95% Confidence Interval: 30.56 to 48.78)) compared to non-liposomal auraptene (50.36 (95% Confidence Interval: 43.58 to 58.19)) (p = 0.0240).

Conclusion: Moreover, the administration of 2.5 and 5 µM of liposomal auraptene led to a reduction in the vessels within the chorioallantoic membrane at the injection site when compared to the control group.

In summary, the use of biodegradable nanoliposomal carriers improved the solubility, release profile, and stability of auraptene while demonstrating anticancer and anti-angiogenic properties.

Background and Aim: Incisional hernia is one of the most notable surgical complications that can be repaired either laparoscopically or by open surgery. This study aims to compare the surgery- related factors and surgical outcomes between these two groups.

Methods: This is a retrospective single center study, investigates the surgical outcomes of patients that have underwent either open or laparoscopic surgical repair in a tertiary hospital in Tehran, between 2019- 2020.

Results: 70 patients (35 in each group) enrolled the study. 71.42 % of patients were female and the mean age of total study sample was 53.12 ± 11.66 years. There were significant lower pain score, operation time, and hospitalization length in laparoscopic cohort (p< 0.05). There was no significant difference between rates of post- operative complications including seroma, hematoma, surgical site infection, and ileus (p> 0.05). Laparoscopic surgery significantly costs more than open surgery (p< 0.05). There was no case of recurrence within the 12-months after surgery.

Conclusion: In conclusion, while laparoscopic surgery costs more than open surgery, it is associated with significant decrease in pain score, operation time, and hospitalization length. There is no significant difference between post-operative complications including seroma, hematoma, and surgical site infection among two groups.

Monoclonal antibodies (mAbs) have changed diagnostics and therapy due to their high specificity and affinity to the target antigens (Ags). Accurately measuring the affinity of mAbs is critical to understanding their binding properties. It represents the strength of binding between an antibody (Ab) and its target Ag and enables decision making in the development and optimization of these antibodies to improve their efficacy in diagnostics and therapy. Various methods such as the equilibrium dissociation constant, ELISA, surface plasmon resonance (SPR), and microarray-based platforms can be used to determine mAb affinity. The non-competitive ELISA is simple and available method for many laboratories, based on the law of mass action that compares the OD50 of three sigmoidal curves of serial Ab dilutions on plates coated with different Ag concentrations to determine the binding strength between a mAb and its Ag. This protocol provides a step-by-step guide to determining mAb affinity using modified ELISA and enables researchers to make informed decisions about the development and application of mAbs in their respective fields.

Lung is one of the vital organs that get severely affected by cytokine storm and sepsis, leading to the development of acute lung injury (ALI) or acute respiratory distress syndrome (ARDS). Mesenchymal stem cell- derived small extracellular vesicles (MSC- derived sEVs) are one of the therapeutic approaches for ARDS/ ALI caused by sepsis. Apart from sEVs’ ability to carry medication, MSC- derived sEVs also possess anti- inflammatory, tissue repair, and regeneration properties. Targeted drug delivery has been a crucial area of concern in the medical field, specifically for treating lung diseases. To treat lung diseases locally, inhalation of drug products has been developed. Drug delivery by inhalation has emerged as an effective method for local administration of therapeutic agents, with numerous benefits including better efficacy at lower doses and decreased toxicity. Additionally, inhalation administration is a viable option for the systemic distribution of medications due to the lungs' considerable absorption surface and their ability to bypass initial metabolism. Therefore, our hypothesis proposes the inhalation of MSC- derived sEVs as a potential strategy for alleviating acute lung injury induced by sepsis-related cytokine storm. Following the isolation and characterization of these MSC- derived sEVs, they will be administered to an animal model of sepsis via a nebulizer, either in their pure form or loaded with drugs. Several approaches will be employed to evaluate lung functionality, including histological analysis and the measurement of inflammatory and regulatory cytokine levels to assess changes in both the humoral and cellular immune systems.

The promastigotes of Leishmania major (MRHO/IR/75/ER) were grown as previously described [1]. J774A.1 which is a murine macrophage Cell Line was purchased from Pasteur Institute of Iran and cultured using Roswell Park Memorial Institute medium (RPMI-1640) (Biowest, France) enriched with 10% heat-inactivated Fetal Bovine Serum (FBS) (Gibco, USA) with 2 mM L.glutamine (Sigma, Australia) and 1% penicillin/streptomycin (Gibco, USA). 7 × 10⁶ macrophage cells were seeded in a T75 cell culture flask (SPL life sciences, South Korea). After 24 hours, 7 × 10⁷ stationary phase promastigotes of L. major were added to the T75 flask containing J774A.1 macrophage cells. The video was recorded immediately and 3 hours after the infection of macrophages with L. major parasites under an inverted microscope (OPTIKA, Italy) with Samsung A13 mobile phone camera (Samsung, USA).

Dendritic cells (DCs) are remarkable professional antigen-presenting cells (APCs) that are pivotal in bridging the gap between innate and adaptive immunity. Given this unique ability, these cells are a promising target for immunotherapy of various diseases. Monocytes serve as a valuable resource for generating DCs in vitro. To generate monocyte-derived DCs and investigate morphological changes during the differentiation phenomenon, following the reception of written informed consent, peripheral blood samples were obtained from healthy donors using falcon tubes containing heparin. Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll solution with a density of 1.077 g/ml and density gradient centrifugation. Monocytes were subsequently extracted from PBMCs through the magnetic-activated cell sorting method (MACS) following the instructions provided with the kit. The culture of monocytes was carried out in a concentration of 1.5 × 10⁶/mL in a 6-well plate using complete culture media (RPMI-1640 supplemented with 15% FBS, 100 µg/mL streptomycin, 100 IU/mL penicillin, and 2 mM L-glutamine), in conjunction with 50 μM 2-mercaptoethanol solution and recombinant human GM-CSF and IL-4 cytokines at concentrations of 40 and 25 ng/mL, respectively. The plate was incubated at 37°C with 5% CO2 and humidification. After three days, half of the culture media was substituted with a new mixture containing GM-CSF and IL-4 cytokines. Immature DC (iDCs) cells were collected on the fifth day. Following this, iDCs were treated with 100 ng/mL of lipopolysaccharide (LPS) and incubated for 24 hours to induce the maturation of iDCs. On the sixth day, mature DCs were harvested. Monocytes and DCs were examined for their morphology, with images captured utilizing an inverted light microscope. In this regard, microscopic analysis revealed morphological alterations between monocytes derived from PBMCs at the initiation of culture and differentiated mature DCs acquired on day 6. Monocytes appeared as round cells with no visible dendrites, in contrast to DCs which were distinguishable by their visible dendrites (Figure1).