Immunoliposomes bearing lymphocyte activation gene 3 fusion protein and P5 peptide: A novel vaccine for breast cancer

LAG3-Ig as an immune adjuvant has elicited potent anti-tumor immune responses in several preclinical and clinical studies, but the full potential immunostimulatory of LAG3-Ig has yet to be achieved. We hypothesized that by anchoring LAG3-Ig to the surface of liposomes, the adjuvant activity of LAG3-Ig could be improved. We also investigated the immunotherapy by co-delivery of liposome-coupled LAG3-Ig and P5 tumor antigen in mice model of TUBO breast cancer. We prepared and characterized novel PEGylated liposomes bearing surface conjugated LAG3-Ig and P5. Consistent with our hypothesis, liposomes-conjugated LAG3-Ig via multivalent binding to MHC class II molecules exerted immunostimulatory of LAG3-Ig and markedly induced maturation of dendritic cells more efficiently than free LAG3-Ig. LAG3-Ig-P5-immunoliposomes effectively elicited protective anti-tumor responses more than locally injected soluble LAG3-Ig + P5. The higher percentage of CD4⁺ and CD8⁺ T cells in the spleen and more rapid and pronounced infiltration of these effector cells into the site of the tumor were seen following immunoliposome therapy. Finally, anti-tumor immunity induced by LAG3-Ig-P5-immunoliposomes translated into the more tumor regression and prolonged survival of treated mice, compared to soluble immunotherapy. Taken together, our findings suggest that LAG3-Ig-P5-immunoliposomes can be considered as a valuable candidate for developing a liposome-based therapeutic cancer vaccine in treating HER2/ neu⁺ breast cancer patients.     

Essential Oil and Bioactive Compounds Variation in Myrtle (Myrtus communis L.) as Affected by Seasonal Variation and Salt Stress

The effect of different NaCl concentrations (control, 2, 4 and 6 dS/m) and three harvesting times in different seasons including spring (9 April), summer (5 July), and fall (23 September) was evaluated on essential oil (EO) yield, composition, phenolic, flavonoid content, and antioxidant activity of myrtle. Essential oil yield ranged from 0.2% in control and fall to 1.6% in moderate salinity (4 dS/m) and spring season. The main constituents obtained from gas chromatography/mass spectrometry analysis were α‐pinene, 1,8‐cineole, limonene, linalool, α‐terpineol, and linalyl acetate in which α‐pinene ranged from 11.70% in moderate and fall to 30.99% in low salinity (2 dS/m) and spring, while 1,8‐cineole varied from 7.42% in high salinity (6 dS/m) and summer to 15.45% in low salinity and spring, respectively. Salt stress also resulted in an increase in total phenolic, flavonoid content, and antioxidant activity. The highest antioxidant activity based on DPPH radical scavenging activity, reducing power (FTC) and β‐carotene/linoleic acid model systems was found in plants harvested in spring and summer in high stress condition. The lowest IC₅₀ values obtained in 6 dS/m in spring (375.23 μg/ml) followed by summer (249.41 μg/ml) and fall (618.38 μg/ml). Eight major phenolic and flavonoid compounds were determined in three harvesting times using high performance liquid chromatography analysis. In overall, late harvesting time of myrtle in fall can lead to reduce the most of major EO components, while it can improve the amount of phenolic acids.     

The ENPP1 K121Q polymorphism is not associated with type 2 diabetes and related metabolic traits in an Iranian population

The K121Q polymorphism of the ectoenzyme nucleotide pyrophosphate phosphodiesterase 1 (ENPP1) gene has been variably associated with insulin resistance and type 2 diabetes (T2D) in several populations. However, this association has not been studied in Iranian subjects and we hypothesized that the K121Q variant might be associated with T2D and related metabolic traits in this population. The K121Q genotypes were determined by PCR-restriction fragment length polymorphism in 377 normoglycemic controls and 155 T2D patients. T2D patients had significantly higher values for systolic and diastolic blood pressure, BMI, glucose, cholesterol, triglyceride, LDL, apoB, insulin, and HOMA-IR, and lower levels of HDL than the normoglycemic subjects. The frequency of the Q allele did not differ between T2D and normoglycemic subjects (OR 0.96, 95% CI 0.90-2.00, P?=?0.70). The Q allele frequency was 16.5% in T2D and 15.2% in normoglycemic subjects. The ENPP1 genotype (KQ?+?QQ) was not associated with the systolic and diastolic blood pressure, glucose, triglyceride, cholesterol, LDL-C and HDL-C, apo B, BMI, HOMA-IR, and insulin levels in both normoglycemic and T2D groups. Our results suggest that the ENPP1 121Q allele might not be associated with T2D and related metabolic traits among Iranian subjects.     

Accuracy evaluation of distance inverse square law in determining virtual electron source location in Siemens Primus linac

Aim

The aim of this study is to evaluate the accuracy of the inverse square law (ISL) method for determining location of virtual electron source (S_Vir) in Siemens Primus linac.

Molecular characterization of Cryptosporidium isolates from humans and animals in Iran

Isolates of Cryptosporidium spp. from human and animal hosts in Iran were characterized on the basis of both the 18S rRNA gene and the Laxer locus. Three Cryptosporidium species, C. hominis, C. parvum, and C. meleagridis, were recognized, and zoonotically transmitted C. parvum was the predominant species found in humans.     

Induction and comparison of different in vitro morphogenesis pathways using embryo of cumin (<Emphasis Type="Italic">Cuminum cyminum</Emphasis> L.) as a model material

In this study, using cumin embryo as explant and manipulating plant growth regulators (PGRs) in regeneration medium, the main in vitro morphogenesis pathways including direct shoot organogenesis, direct somatic embryogenesis, indirect somatic embryogenesis, and indirect shoot organogenesis were obtained. The effects of PGRs, subculture, and light on the induction and progression of different pathways were studied in detail. Direct shoot organogenesis occurred on the meristematic zone, while direct somatic embryogenesis was observed on hypocotyl part of cumin embryo (more differentiated part). Application of BAP (0.1 mgl⁻¹) was the sole triggering factor for induction of callus and indirect regeneration pathways. Exogenous IAA played the central role in the direct somatic embryogenesis pathway; however, the combined effects of IAA and NAA along with the high endogenous cytokinin level resulted in direct shoot organogenesis. Subculturing revealed accelerating effects on direct somatic embryogenesis pathway and callus formation. Conversely, subculturing had negative effect on direct shoot organogenesis pathway. In certain combinations of PGRs, like 0.4 mgl⁻¹ IAA + 0.4 mgl⁻¹ NAA, co-induction and co-regeneration of different pathways were observed. Investigation of genotype dependencies of different pathways showed that direct pathways are more genotype-dependent, stable, and faster than indirect pathways. This research presents the embryo of cumin as a convenient model material for induction and comparison of different morphogenesis pathways.     

Intestinal pH profile in rainbow trout Oncorhynchus mykiss and microhabitat preference of the flagellate Spironucleus salmonis (Diplomonadida)

In farmed rainbow trout Oncorhynchus mykiss, the flagellate Spironucleus salmonis (Diplomonadida) is often found in the pyloric region of the intestine. While previous in vitro studies report a pH of 7.5 to 8.0 as optimal for presumed S. salmonis, no previous in vivo studies have investigated the relationship between pH and microhabitat preference. Therefore, in 698 rainbow trout (75% were 5 to 6 mo old juveniles, 10 to 20 cm total length), we recorded occurrence and density of S. salmonis, and pH, in the pyloric, anterior, middle, and posterior intestine. There were no significant differences in total length or weight between infected and uninfected fish. S. salmonis preferred the pyloric region, with occurrence and density decreasing significantly from pyloric to posterior regions. In infected fish, pH in pyloric (6.8 to 7.9, mean 7.3) and posterior regions (6.5 to 8.0, mean 7.1) was significantly lower than in anterior (6.5 to 8.5, mean 7.7) and middle (6.8 to 8.2, mean 7.7) regions; in uninfected fish, the pH profile was similar. At the individual level, 90 % of infected fish and 79% of uninfected fish showed this pH profile. In the pyloric region, pH was not significantly different among uninfected fish, and fish with light, moderate, or heavy infections. Our in vivo study suggests the optimal pH for S. salmonis is between 7.1 and 7.5, possibly close to 7.3 (the mean in pyloric region of infected fish). We conclude that while the presence of S. salmonis reflected tolerable pH, density of infection was not correlated with pH, and thus a causal relationship between microhabitat preference and pH is unlikely.     

Suppression of islet allogeneic immune response by indoleamine 2,3 dioxygenase-expressing fibroblasts

Success of transplantation of pancreatic islets which is a promising way for restoring efficient insulin regulation in type 1 diabetes depends on lifelong use of immunosuppressive drugs. To eliminate the use of systemic immunosuppressive drugs for islet transplantation, we examined the potential use of a local immunosuppressive factor, indoleamine 2,3-dioxygenase (IDO). Thus, the aim of this study was to determine whether local expression of IDO in bystander syngeneic fibroblasts could prevent islet allogeneic immune response in vitro. C57BL/6 (B6) mouse fibroblasts were induced to express IDO by either IFN-gamma treatment or transduction with an adenoviral vector and were co-cultured with B6 mouse lymphocytes and BALB/c mouse pancreatic islets in the presence or absence of an IDO inhibitor. Proliferation of lymphocytes were then assessed using [(3)H]-thymidine incorporation assay. IDO-expression by co-cultured syngeneic fibroblasts resulted in a five-fold decrease in lymphocyte proliferation rate upon stimulation of lymphocytes by allogeneic mouse pancreatic islets (21.9% +/- 5.3 and 22.1% +/- 4.9 in the preparations with IFN-gamma treated and genetically modified IDO-expressing fibroblasts, respectively vs. 100% in control groups, P < 0.01). Allogeneic response was restored when IDO inhibitor was added to the culture indicating that suppression was due to IDO. In conclusion, this study shows that local expression of IDO by syngeneic bystander fibroblasts can suppress in vitro proliferation of lymphocytes in response to stimulation with allogeneic pancreatic islets. This local immunosuppressive function of IDO may be employed for development of a novel alternative strategy for preventing allogeneic islet graft rejection.