Comprehensive genetic discrimination of Leonurus cardiaca populations by AFLP,ISSR, RAPD and IRAP molecular markers

Leonurus cardiaca is well known for its medicinal importance. In this investigation, genotypic characterization of this species from six eco-geographical regions of Iran was evaluated by four molecular techniques (AFLP, RAPD, ISSR and IRAP). A total of 899 polymorphic fragments were detected by used molecular markers (AFLP = 356, RAPD = 325, ISSR = 113 and IRAP = 105) with an overall average polymorphism of 81.24 %. Genetic variation calculated using Shannon’s Information index (I) and Nei’s gene diversity index (H) showed high genetic diversity in studied germplasm. Also, analysis of molecular variance showed high genetic variation among (55 %) and within populations (45 %). UPGMA dendrogram constructed from combined data of molecular markers distinguished studied populations in accordance with the results obtained by each marker which all individuals were clearly differentiated into two major clusters. The correlation coefficients were statistically significant for all marker systems with the highest correlation between similarity matrixes of RAPD and ISSR markers (r = 0.82). The present results have an important implication for L. cardiaca germplasm characterization, improvement, and conservation. Furthermore, the characterized individuals exhibited a great deal of molecular variation and they seem to have a rich gene pool for breeding programs.     

Multivariate analysis of motherwort germplasm in Iran using morphological variables and essential oil content

The morphological variation and essential oil content of 150 individuals of motherwort (Leonurus cardiaca) from six natural regions of Iran were examined biometrically based on multivariate analysis. The highest variation was observed in plant height, length of main stems, main inflorescence length, floral cycles of main stem, lateral branches length, basal leaf length, floral leaf length width, flower length, and calyx color. Results of simple correlation analysis showed the existence of significant positive and negative correlations among some important parameters. The highest correlation was observed between plant height and length of main stems and between lateral branch length and lower surface color of leaf. Populations were clustered in two groups representing subspecies cardiaca and persicus. The subspecies cardiaca populations were closely related with each other and differentiated from the subspecies persicus by pubescence stems and leaves, more number of floral cycles, higher compaction of floral cycles, lower number of lateral branches, shorter length of petioles and leaves and smaller floral leaves. Essential oil yields varied from 0.02 (for Taleghan population) to 0.053 mg/100 g dry mater (for Kerman population). The conservation of the highly diverse native populations of Iranian motherwort germplasm is recommended.     

Cost-of-illness analysis of type 2 diabetes mellitus in Iran

Introduction

Diabetes is a worldwide high prevalence chronic progressive disease that poses a significant challenge to healthcare systems. The aim of this study is to provide a detailed economic burden of diagnosed type 2 diabetes mellitus (T2DM) and its complications in Iran in 2009 year.

Methods

This is a prevalence-based cost-of-illness study focusing on quantifying direct health care costs by bottom-up approach. Data on inpatient hospital services, outpatient clinic visits, physician services, drugs, laboratory test, education and non-medical cost were collected from two national registries. The human capital approach was used to calculate indirect costs separately in male and female and also among different age groups.

Results

The total national cost of diagnosed T2DM in 2009 is estimated at 3.78 billion USA dollars (USD) including 2.04±0.28 billion direct (medical and non-medical) costs and indirect costs of 1.73 million. Average direct and indirect cost per capita was 842.6±102 and 864.8 USD respectively. Complications (48.9%) and drugs (23.8%) were main components of direct cost. The largest components of medical expenditures attributed to diabetes''s complications are cardiovascular disease (42.3% of total Complications cost), nephropathy (23%) and ophthalmic complications (14%). Indirect costs include temporarily disability (335.7 million), permanent disability (452.4 million) and reduced productivity due to premature mortality (950.3 million).

Conclusions

T2DM is a costly disease in the Iran healthcare system and consume more than 8.69% of total health expenditure. In addition to these quantified costs, T2DM imposes high intangible costs on society in terms of reduced quality of life. Identification of effective new strategies for the control of diabetes and its complications is a public health priority.     

A comparative study on the allergenicity, structure, and ultrastructure of two Acacia pollen grains in guinea pigs

Acacia saligna and Acacia victoria are native to Australia brought to and grown in Southern Iran. They have since had a widespread distribution and become native in Iran. As people of the region are exposed to the pollen grains from these plants during a 5 months period, this study aimed at investigating the allergenicity of their mature pollen grains. In addition, the structural and ultrastructural as well as the total protein content of the mature pollen grains were analyzed. Pollen grains of A. saligna and A. victoria were collected from the suburbs of Ahvaz, Iran. Pollen extracts (15 % concentration) were prepared in PBS (pH 7.2), and 4- to 6-week-old male guinea pigs (Hartly) were used for allergenicity tests. Skin tests showed a significant increase in flare diameter. Clinical tests also showed a significant change in the levels of eosinophils, neutrophils, and IgE. Comparative analysis of the allergenicity of A. saligna and A. victoria pollen grains revealed a higher IgE level in the latter. Comparative analysis of pollen characteristics was performed using both light and electron microscopy for sporoderm structure and ultrastructure, respectively. Twelve percent PAGE analysis of the total protein content of mature pollen grains showed a greater number of bands in A. victoria, while four bands were common in both species. In conclusion, this study demonstrates that both A. saligna and A. victoria pollen grains are allergenic, with a greater allergenicity of A. victoria pollen grains. In addition, the structural and ultrastructural as well as the total protein content of the mature pollen grains are revealed, and a potential protein allergen is proposed.     

The fauna and distribution of house dust mites in residential homes of Bandar Abbas District,Southern Iran

This study was designed to determine the occurrence, distribution and abundance of house dust mites (HDM) in residential homes in Bandar Abbas (Hormozgan Province), because of numerous complaints of allergies in this oriental city. The study area was divided in five sampling zones based on population density and geographical distribution. In each sampling zone 10 houses were randomly selected. A total of 50 home dust samples were collected using a portable vacuum cleaner for 2 min from 1 m² of the surface of mattresses, carpets, sofas and furniture in residential houses. After collection, samples were immediately frozen. Mite species were identified and counted using standard methods and keys. Of the sampled houses 88% (44 houses) were contaminated with at least one HDM species. Three species were identified: Dermatophagoides pteronyssinus (63.1%), D. farinae (32.8%) and D. evansi (4.1%) (Pyroglyphidae). Our findings indicate a relationship between HDM density and moisture and temperature of residential places. The high contamination rate of residential houses (88%) and the favourable environmental conditions for these arthropods stress that they should be considered as important allergic causing agents.     

Identification and validation of Asteraceae miRNAs by the expressed sequence tag analysis

MicroRNAs (miRNAs) are small non-coding RNA molecules that play a vital role in the regulation of gene expression. Despite their identification in hundreds of plant species, few miRNAs have been identified in the Asteraceae, a large family that comprises approximately one tenth of all flowering plants. In this study, we used the expressed sequence tag (EST) analysis to identify potential conserved miRNAs and their putative target genes in the Asteraceae. We applied quantitative Real-Time PCR (qRT-PCR) to confirm the expression of eight potential miRNAs in Carthamus tinctorius and Helianthus annuus. We also performed qRT-PCR analysis to investigate the differential expression pattern of five newly identified miRNAs during five different cotyledon growth stages in safflower. Using these methods, we successfully identified and characterized 151 potentially conserved miRNAs, belonging to 26 miRNA families, in 11 genus of Asteraceae. EST analysis predicted that the newly identified conserved Asteraceae miRNAs target 130 total protein-coding ESTs in sunflower and safflower, as well as 433 additional target genes in other plant species. We experimentally confirmed the existence of seven predicted miRNAs, (miR156, miR159, miR160, miR162, miR166, miR396, and miR398) in safflower and sunflower seedlings. We also observed that five out of eight miRNAs are differentially expressed during cotyledon development. Our results indicate that miRNAs may be involved in the regulation of gene expression during seed germination and the formation of the cotyledons in the Asteraceae. The findings of this study might ultimately help in the understanding of miRNA-mediated gene regulation in important crop species.     

Using Maize (Zea mays L.) and Sewage Sludge to Remediate a Petroleum-Contaminated Calcareous Soil

Phyto-stimulation, the use of plants to stimulate activity of microorganisms in a root zone, has been proposed as an approach to promote the degradation of petroleum hydrocarbons and thus the remediation of petroleum-polluted soils. In this study, we investigated the potential use of sewage sludge to enhance phyto-stimulating effects of maize (Zea mays L.) on the elimination of an aged petroleum contamination in a calcareous soil. In a pot experiment, maize was grown on the experimental soil for two months at three levels of sewage sludge application (0, 20, and 50 g dry matter of sludge per kg soil). The amendments increased root and shoot growth of the experimental plants approximately by a factor of two at the lower sludge treatment level and by a factor of five at the higher sludge treatment level. In a separate incubation experiment, sludge application also led to an immediate stimulation of soil respiration, which then further increased over time. The initial stimulation was three times larger at the higher than at the lower treatment level, but the rate of subsequent increase was similar in both treatments. The two sludge treatments also accelerated TPH elimination in the contaminated soil, and again the effect was approximately three times stronger at the higher than at the lower treatment level. The sludge effect on TPH elimination was much stronger than the effect of the plants. More than half of the initial contamination was reduced in combined treatment with maize and sludge application at the highest rate. The results show that sewage sludge can substantially enhance the remediation of petroleum-contaminated soil, especially when applied in conjunction with a suitable plant such as maize.     

A study on the allergenicity and ontogeny of Acacia farnesiana pollen grains in guinea pigs

Pollen grains as the angiosperm male gametophytes transfer male genetic material during sexual reproduction. Pollen grains are among the important plant allergens, such that almost 80–90 % of plant allergens are of pollen origin. Acacia farnesiana is a plant with economical values, and due to its resistance to dry climates, it has had a widespread distribution in Southern Iran. This study was aimed at investigating the allergenicity and the ontogeny of pollen grains of A. farnesiana. Pollen grains were collected from the blossoms and flowers of A. farnesiana in the suburbs of Ahvaz-Iran. Pollen extracts (15 %) were prepared in PBS (pH 7.2). 4–6-week-old male guinea pigs (Hartly) were used for allergenicity tests. Skin tests showed a significant increase in flare diameter. Clinical tests also showed a significant change in the levels of eosinophils, neutrophils, and IgE. Histotechnical analysis was performed on male gametophytes, and photographs were taken using a camera-equipped light microscope. Pollen characteristics were identified using both light and electron microscopy. Sporoderm ultrastructure and pollen morphology were studied using Scanning Electron Microscopy. PAGE analysis of the total protein content of mature pollen grains showed seven clear bands of 10–83 kDa.     

Osmolytic Effect of Sucrose on Thermal Denaturation of Pea Seedling Copper Amine Oxidase

Protein stability is a subject of interest by many researchers. One of the common methods to increase the protein stability is using the osmolytes. Many studies and theories analyzed and explained osmolytic effect by equilibrium thermodynamic while most proteins undergo an irreversible denaturation. In current study we investigated the effect of sucrose as an osmolyte on the thermal denaturation of pea seedlings amine oxidase by the enzyme activity, fluorescence spectroscopy, circular dichroism, and differential scanning calorimetry. All experiments are in agreement that pea seedlings amine oxidase denaturation is controlled kinetically and its kinetic stability is increased in presence of sucrose. Differential scanning calorimetry experiments at different scanning rates showed that pea seedlings amine oxidase unfolding obeys two-state irreversible model. Fitting the differential scanning calorimetry data to two-state irreversible model showed that unfolding enthalpy and T ^*, temperature at which rate constant equals unit per minute, are increased while activation energy is not affected by increase in sucrose concentration. We concluded that osmolytes decrease the molecular oscillation of irreversible proteins which leads to decline in unfolding rate constant.