Expression of an innate immune element (mouse hepcidin-1) in baculovirus expression system and the comparison of its function with synthetic human hepcidin-25

Hepcidin is an innate immune element which decreases the iron absorption from diet and iron releasing from macrophage cell. In contrast to the chemical iron chelators, there has been limited effort applied to the specific use of hepcidin as a new drug for decreasing the iron overload. Hepcidin is produced in different biological systems. For instance, E-coli is used for human hepcidin expression, however, post-translational modification is impaired. We have used a simple baculovirus expression system (BES) to improve the hepcidin folding and activity. Hepcidin Messenger Ribonucleic acid (mRNA) was isolated from mouse liver cells and its complementary Deoxyribonucleic acid (cDNA) was produced and amplified. PFastBac HTB vector was used for recombinant bacmid production. Recombinant baculovirus was produced using SF-9 cell line. The mouse hepcidin-1 protein was expressed in a large quantity and functional tests were performed for this recombinant peptide. The yield of hepcidin in BES was 20 μg/mL and anti-histidine (anti-His) tag antibody was used for the confirmation of hepcidin on western blot nitrocellulose paper. Functional tests showed that mouse hepcidin accumulates iron in the macrophage cell line J774A.1 up to 63%. In addition, our data showed that the mouse hepcidin-1 has less toxicity compared to the synthetic human hepcidin-25 (p = 0.000). © 2011 by School of Pharmacy

The incidence of prostate cancer in Iran: Results of a population-based cancer registry

Background: Little is known about the epidemiology of prostate cancer in Iranian men. We carried out an active prostate cancer surveillance program in five provinces of Iran. Methods: Data used in this study were obtained from population-based cancer registries between 1996 and 2000. Results: The age-standardized incidence rate of prostate carcinoma in the five provinces was 5.1 per 100,000 person-years. No significant difference was seen in the age-standardized incidence rate of prostate cancer within the provinces studied. The mean±SD age of patients with prostate cancer was 67±13.5 years. Conclusion: The incidence of prostate cancer in Iran is very low as compared to the Western countries. This can partly be explained by lack of nationwide screening program, younger age structure and quality of cancer registration system in Iran

Hydrogel nanoparticle encapsulated plasmid as a suitable gene delivery system

To facilitate the delivery of genetic material, the use of appropriate carriers such as polymers is necessary. Nanoparticles comprising of chitosan-alginate polymers were formed through pregel preparation method. Chi/Alg nanoparticles had a mean Z-Average diameter of 161.8 nm and mean zeta 29.3 mV, respectively. The ability of plasmid-complex in preventing DNA migration showed Chi/Alg nanoparticles have great capacity to maintain plasmid. The efficiency of nanoparticles for transfection of pEGFP-N1 plasmid in the cultured HEK 293 cells was measured by flow cytometry. Cell viability assays indicated that nanoparticles had no toxic effect on HEK 293 cells after 4 or 24 h. Our suitable candidate for gene delivery would be alg/chi nanoparticles.Для облегчения доставки генетического материала необходимо использование подходящих носителей, таких как полимеры. Наночастицы, состоящие из хитозан-альгинатных полимеров, были получены методом подготовки прегеля. Chi/Alg наночастицы имели средний диаметр 161.8 нм (Z-Average) и средний zeta-потенциал 29.3 mV. Отсутствие миграции ДНК во время электрофореза комплексов плазмиды с наночастицами показало, что Chi/Alg наночастицы могут удерживать плазмидную ДНК внутри комплекса. Эффективность наночастиц для трансфекции плазмиды pEGFP-N1 в культивируемые клетки HEK 293 была измерена с помощью жидкостной цитометрии. Тесты на жизнеспособ-ность клеток показали, что наночастицы не имели токсичного эффекта на клетки HEK 293 через 4 ч или 24 ч. Наночастицы Alg/Chi являются подходящим кандидатом для доставки генов

The Problem of Mixing up of Leishmania Isolates in the Laboratory: Suggestion of ITS1 Gene Sequencing for Verification of Species

Background: Leishmaniasis is endemic in Iran. Different species of Leishmania (L.) parasites are causative agents of this disease. Correct identification of Leishmania species is important for clinical studies,prevention, and control of the diseases. Mix up of Leishmania isolates is possible in the laboratory, so there is need for verification of species for isolates of uncertain identity. Different methods may be used for this purpose including isoenzyme electrophoresis and molecular methods. The isoenzyme lectrophoresis, due to its drawbacks, is feasible only in specialized laboratories while molecular methods may be more feasible. The aim of this research was to study the application of the internal transcribedspacer 1 (ITS1) sequencing method, in comparison to isoenzyme electrophoresis method, for verification of Leishmania species.Methods: Six Leishmania isolates were received from different research institutions in Iran. The species of these isolates were known by donating institution according to their isoenzyme profile. The species of these isolates were re-identified in Pasteur Institute of Iran by PCR amplification of ITS1 followed bysequencing and comparison of these sequences with Leishmania sequences in GenBank. Isoenzyme electrophoresis was performed for confirmation of the results of ITS1.Results: ITS1 sequence showed that some isolates were mixed up or contaminated with Crithidia. Isoenzyme electrophoresis confirmed the results of ITS1 sequences.Conclusion: ITS1 sequencing is relatively more feasible than the traditional isoenzyme electrophoresismethod and is suggested for verification of Leishmania species

Molecular Cloning and Characterization of P4 Nuclease from Leishmania infantum

Parasite of the genus Leishmania is reliant on the salvage pathway for recycling of ribonucleotides. A class I nuclease enzyme also known as P4 nuclease is involved in salvage of purines in cutaneous Leishmania species but the relevant enzymes have not been characterized in Leishmania infantum (L. infantum). The aim of this study was to clone and characterize the gene encoding class I nuclease in L. infantum. DNA extracted from L. infantum was used for amplification of P4 nuclease gene (Li-P4) by PCR. The product was cloned, sequenced, and expressed in E. coli for further characterization. Analysis of the sequence of Li-P4 revealed that the gene consists of an ORF of 951 bp. Sequence similarity analysis indicated that Li-P4 has a high homology to relevant enzymes of other kintoplastids with the highest homology (88%) to p1/s1 class I nuclease from L. donovani. Western blotting of antirecombinant Li-P4 with promastigote and amastigote stages of L. infantum showed that this nuclease is present in both stages of parasite with higher expression in amastigote stage. The highly conserved nature of this essential enzyme in Leishmania parasites suggests it as a promising drug target for leishmaniasis

Multimorbidity as an important issue among women: results of gender difference investigation in a large population-based cross-sectional study in West Asia

Objectives: To investigate the impact of gender on multimorbidity in northern Iran. Design: A cross-sectional analysis of the Golestan cohort data. Setting: Golestan Province, Iran. Study population: 49 946 residents (age 40–75 years) of Golestan Province, Iran. Main outcome measures: Researchers collected data related to multimorbidity, defined as co-existence of two or more chronic diseases in an individual, at the beginning of a representative cohort study which recruited its participants from 2004 to 2008. The researchers utilised simple and multiple Poisson regression models with robust variances to examine the simultaneous effects of multiple factors. Results: Women had a 25.0% prevalence of multimorbidity, whereas men had a 13.4% prevalence (p<0.001). Women of all age-groups had a higher prevalence of multimorbidity. Of note, multimorbidity began at a lower age (40–49 years) in women (17.3%) compared with men (8.6%) of the same age (p<0.001). This study identified significant interactions between gender as well as socioeconomic status, ethnicity, physical activity, marital status, education level and smoking (p<0.01). Conclusion: Prevention and control of multimorbidity requires health promotion programmes to increase public awareness about the modifiable risk factors, particularly among women

Development of monoclonal antibodies against axenic amastigotes of Leishmania infantum strain in Iran: Implication for diagnosis of Kala-azar

Objective(s): Leishmaniasis is endemic in 88 countries. Amastigote forms of Leishmania are experts at exploiting host cell processes to establish infection. Monoclonal antibodies are key reagents used in the diagnosis of infectious and non-infectious diseases. The aim of this study was to produce monoclonal antibodies against axenic amastigotes of the Leishmania infantum strain in Iran.Materials and Methods: First, standard strains were cultured and axenic amastigote antigens of L. infantum were obtained. Since then, BALB/c mice were immunized and antibody titers were determined. For hybridoma cell formation, lymphocytes isolated from spleen of immunized mice and myeloma cells were fused at a ratio of 10 to 1 in the presence of polyethylene glycol, followed by limiting dilution for the isolation of monoclones. Subsequently, antibody isotypes were determined by using the isotyping kit. The best clone was injected intraperitoneally to pristane-primed mice for large scale production of monoclonal antibodies. The specificity of antibody was determined with Western blotting.Results: Approximately 25 positive monoclones were obtained, of which four hybrids producing anti-amastigotes L. infantum monoclonal antibodies with high optical density (OD), selected and designated as 8D2 FVI6, 8D2 FVI3, 6G2 FV4 and 6G2 FV3. Results from isotype determination showed the IgG2b sub-class in 6G2FV2 and 8D2FVI6 monoclones. Conclusion: This study produced monoclonal antibody against amastigotes of Iranian strain of L. infantum for the first time. These antibodies have reactivity against Iranian strain of L. infantum and can be used in the diagnosis of Kala-azar

Adjuvant Effect of Leishmania major Promastigotes on the Immune Response of Mice to Ovalbumin

ABSTRACT The immune responses of mice immunized with ovalbumin (OVA) together with killed L. major (KLM) promastigotes as adjuvant were studied. Three doses (5 × 10 7 , 1 × 10 8 and 2 × 10 8 ) of KLM combined with OVA (100 µg) were injected into the groups of C57BL/6 mice. BCG and complete Freund's adjuvant (CFA) were used as control adjuvants. Lymphocyte proliferation and antibody titers were determined, and IFN-γ and IL-4 were measured in the supernatants of lymph node cell cultures. Results showed that immunization using OVA mixed with KLM enhanced the in vitro proliferative response of T-cells to the antigen and resulted in the production of increased levels of IFN-γ (2800-3700 pg/ml) relative to the mice injected with OVA alone (1750 pg/ml). In the mice receiving OVA + 5 × 10 7 KLM, the production of IL-4 remained lower (18, 20 pg/ml) than OVA alone (105, 109 pg/ml) and almost was similar to that of observed in mice inoculated with OVA + BCG, leading to high IFN-γ/IL-4 ratios. Using higher doses of KLM (1 × 10 8 ), the IL-4 responses were of the same magnitude as or higher than the responses of mice inoculated with OVA + CFA. Antibody titers to OVA were also strongly boosted at the highest KLM dose. These findings indicate that KLM may function as an adjuvant, and its dose plays a role in the eventual outcome of the response. Inoculation of the mice with a low dose of KLM (5 × 10 7 ) tends to promote a Th1-type response. Iran. Biomed. J. 6 (4): 123-128, 200

Late Pleistocene and Holocene sea-level change and coastal paleoenvironment evolution along the Iranian Caspian shore

© The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).The level of the Caspian Sea is influenced by rivers mostly from the high latitudes of the Northern hemisphere and therefore any change of its catchments including temperature and precipitation directly reflects on Caspian Sea-level. We reconstructed Late Pleistocene to Holocene Caspian Sea-level by a multi-disciplinary approach from a 27.7m long core in the SE corner of the Iranian Caspian coast in the Gomishan Lagoon. Late Pleistocene deposits containing typical Pleistocene fauna and dated around 20,120 cal yr BP bordered with a major hiatus indicating sea-level fall. Lagoonal deposits with shells dated at around 10,590 cal yr BP suggest that, after this deep lowstand, an initial transgression started, leading to landward advance of barrier–lagoon systems which still continued without any lowstand until 8400 cal yr BP. This corresponded to a biofacies change from lagoonal to the deeper biofacies including diatom and Gastropoda species. Around 8400 cal yr BP sea-level started to fall again, and reddish oxidized sediments with abundant foraminifera (Ammonia beccarii) record a regressive phase around 7700 cal yr BP. The mid-Holocene between 15.7 and 4.9 depths is characterized by a shallow marine environment mostly with high carbonate and gypsum contents, and lagoonal and highstand tract with no subaerial facies. The upper part of the core above a 4.9 m depth reflects at least five Late Holocene Caspian Sea-level cycles from 3260 cal yr BP onward. The Caspian Sea-levels are influenced both by global and regional events.The Oceanography Institute and Cultural Heritage Tourism Organization of Mazandaran

The marginal causal effect of opium consumption on the upper gastrointestinal cancer death using parametric g-formula: An analysis of 49,946 cases in the Golestan Cohort Study, Iran

Upper gastrointestinal (UGI) cancer, including esophageal and gastric, is one of the most common cancers in the world. Hence, the determination of risk factors of UGI helps to reduce the economic and social burden of this cancer in communities. In Iran, the consumption of opium because of its neighborhood with Afghanistan are considerable. In this study, we examine the causal effect of opium use on the time to UGI cancer death. Based on the Golestan Cohort Study (GCS) in northeastern of Iran, about 50000 adults were enrolled to the study for four years (2004–2008) and followed annually until July 2018. We used “parametric g-formula” to study the causal effect of opium use on the time to death due to UGI. In this study, the information of 49946 individuals due to missingness were analyzed. So the median of follow-up time was 144 months and the prevalence of opium use was 17% (about 8489 persons). During the follow-up period, 593 (1.2%) death from upper gastrointestinal cancer were reported. The study showed that the effect of opium use on the time to UGI death was statistically significant (adjusted risk-ratio based on parametric g-formula = 1.31, 95% CI: [1.04, 1.65]). Additionally, the Population Attributable Fraction (PAF) in UGI cancer deaths of opium use was estimated 5.3% (95% CI: [0.6%, 11.3%]). Our results showed a causal effect of opium use on the intensity of upper gastrointestinal cancer deat