The Role of Historical Persian Gardens on the Health Status of Contemporary Urban Residents

The inherent economic and social challenges in major cities have been known to foster stress among the urban population. Frequent stress over long periods may well have serious damaging outcomes, resulting in ailments such as burnout syndrome, sleeplessness and exhaustion, depression, feelings of panic, among others. Therefore, providing access to resources that may enable people to cope with the stress of urban life has become a crucial phenomenon in the twentieth century. Increasing empirical evidence indicates that the presence of natural areas can contribute to enhancing the quality of life in many ways. This study examines two historical Persian gardens from the residents’ perspective in well-known, historic cities of Iran: Isfahan and Kerman. The data were collected through questionnaires (n = 252), semi-structured interviews (n = 20), and visual observation techniques. The findings demonstrate that nature, diversity and the gardens’ historical background, and coherence motivate the residents’ frequent visits to the gardens, which help to address their social, psychological, and physical needs. In addition, the residents’ involvements and the variety of experiences that occur in the gardens lead to the creation of deeper meanings and values associated with the gardens. Subsequently, these construct functional and emotional attachment that evokes a sense of place and identity and may contribute to society’s health and well-being.     

Purification and characterization of a novel extracellular halophilic and organic solvent-tolerant amylopullulanase from the haloarchaeon, Halorubrum sp. strain Ha25

A halophilic archaeon, Halorubrum sp. strain Ha25, produced extracellular halophilic organic solvent-tolerant amylopullulanase. The maximum enzyme production was at high salt concentration, 3–4 M NaCl. Optimum pH and temperature for enzyme production were 7.0 and 40 °C, respectively. Molecular mass of purified enzyme was estimated to be about 140 kDa by SDS–PAGE. This enzyme was active on pullulan and starch as substrates. The apparent K _m for the enzyme activity on pullulan was 4 mg/ml and for soluble starch was 1.8 mg/ml. Optimum temperature for amylolytic and pullulytic activities was 50 °C. Optimum pH for amylolytic activity was 7 and for pullulytic activity was 7.5. This enzyme was active over a wide range of concentrations (0–4.5 M) of NaCl. The effect of organic solvents on the enzyme activities showed that this enzyme was more stable in the presence of non-polar organic solvents than polar solvents. This study is the first report on amylopullulanase production in halophilic bacteria and archaea.     

Flow‐injection chemiluminescence determination of gentamicin: optimization by central composite design

A simple and sensitive flow‐injection chemiluminescence (CL) method has been developed for the determination of gentamicin sulfate. The method is based on the inhibitory effect of gentamicin on the CL emission accompanying oxidation of luminol by H₂O₂ in an alkaline medium in the presence of Cu(II) as a catalyst. Inhibition was caused by the formation of a strong complex between analyte and the catalyst. Experimental variables, including the concentrations of luminol (µmol/L), H₂O₂ (mol/L), Cu(II) (mol/L) and NaOH (mol/L), were optimized using a central composite design. Under optimum conditions, the plot of CL intensity versus gentamicin concentration was found to have two linear ranges. One range was at low concentrations from 1.0 to 10.0 mg/L and the other was from 10.0 to 30.0 mg/L. Precision was calculated by analyzing samples containing 5.0 mg/L gentamicin (n = 11) and the relative standard deviation (RSD) was 1.7%. Also, a high injection throughput of 120 samples/h was achieved. This method was successfully applied to the determination of gentamicin sulfate in pharmaceutical formulations and water samples. Copyright © 2013 John Wiley & Sons, Ltd.     

The Effect of Quercetin Nanosuspension on Prostate Cancer Cell Line LNCaP via Hedgehog Signaling Pathway

Background:Prostate cancer (PCa) is the second leading cause of cancer death in American population. In this manner, novel therapeutic approaches for identification of therapeutic targets for PCa has significant clinical implications. Quercetin is a potent cancer therapeutic agent and dietary antioxidant present in fruit and vegetables.Methods:To investigate the underlying mechanism by which the PCa was regulated, nanoparticles of quercetin were administrated to cells. For in vitro experiments, human PCa cell line LNCaP were involved. Cell viability assay and quantitative RT-PCR (qRT-PCR) for hedgehog signaling pathway genes were used to determine the key signaling pathway regulated for PCa progression.Results:The cell viability gradually decreased with increased concentration of quercetin nanoparticles. At 48 h, 40 mM concentration of quercetin treatment showed near 50% of viable cells. Quercetin nanoparticles upregulates Su(Fu) mRNA expressions and downregulates gli mRNA expressions in the LNCaP cells.Conclusion:The results showed that the hedgehog signaling targeted inhibition may have important implications of PCa therapeutics. Additionally, the outcomes provided new mechanistic basis for further examination of quercetin nanoparticles to discover potential treatment strategies and new targets for PCa inhibition.Key Words: Hedgehog, Prostate cancer, Proliferation, Quercetin nanoparticles, Signaling pathway     

Secondary embryogenesis and transient expression of the β-glucuronidase gene in hypocotyls of rapeseed microspore-derived embryos

Secondary embryogenesis from rapeseed microspore-derived embryos (MDEs) was studied in three Brassica napus L. cultivars Global, PF₇₀₄ and Option. The best results in terms of secondary embryogenesis percentage obtained in cultures of Global and PF₇₀₄ MDEs (75.88 and 65.97 %, respectively) and PF₇₀₄ produced the highest number of secondary embryos per each primary embryo (14.91 ± 2.18). After optimization of physical parameters, rapeseed hypocotyls of MDEs were bombarded with microcarriers coated with a plasmid containing GUS reporter gene. The highest levels of transient GUS expression were obtained using bombardment with gold particles of 1.6 μm, at helium pressure of 9.3 MPa, a bombardment distance of 9 cm, chamber vacuum pressure of 7.1 × 10⁻⁶ kPa and single bombardment in bombardment medium containing 0.4 M mannitol.     

Comparison of Species Richness Estimates Obtained Using Nearly Complete Fragments and Simulated Pyrosequencing-Generated Fragments in 16S rRNA Gene-Based Environmental Surveys

Pyrosequencing-based 16S rRNA gene surveys are increasingly utilized to study highly diverse bacterial communities, with special emphasis on utilizing the large number of sequences obtained (tens to hundreds of thousands) for species richness estimation. However, it is not yet clear how the number of operational taxonomic units (OTUs) and, hence, species richness estimates determined using shorter fragments at different taxonomic cutoffs correlates with the number of OTUs assigned using longer, nearly complete 16S rRNA gene fragments. We constructed a 16S rRNA clone library from an undisturbed tallgrass prairie soil (1,132 clones) and used it to compare species richness estimates obtained using eight pyrosequencing candidate fragments (99 to 361 bp in length) and the nearly full-length fragment. Fragments encompassing the V1 and V2 (V1+V2) region and the V6 region (generated using primer pairs 8F-338R and 967F-1046R) overestimated species richness; fragments encompassing the V3, V7, and V7+V8 hypervariable regions (generated using primer pairs 338F-530R, 1046F-1220R, and 1046F-1392R) underestimated species richness; and fragments encompassing the V4, V5+V6, and V6+V7 regions (generated using primer pairs 530F-805R, 805F-1046R, and 967F-1220R) provided estimates comparable to those obtained with the nearly full-length fragment. These patterns were observed regardless of the alignment method utilized or the parameter used to gauge comparative levels of species richness (number of OTUs observed, slope of scatter plots of pairwise distance values for short and nearly complete fragments, and nonparametric and parametric species richness estimates). Similar results were obtained when analyzing three other datasets derived from soil, adult Zebrafish gut, and basaltic formations in the East Pacific Rise. Regression analysis indicated that these observed discrepancies in species richness estimates within various regions could readily be explained by the proportions of hypervariable, variable, and conserved base pairs within an examined fragment.Culture-independent 16S rRNA gene surveys are now routinely utilized to examine the microbial diversity in various environmental habitats. However, in surveys of highly diverse ecosystems, the size of clone libraries typically constructed (100 to 500 clones) allows for the identification only of members of the community that are present in high abundance (2, 13, 14, 17, 24, 51). In addition to the failure to detect the rare members of the ecosystem, these relatively small datasets provide inaccurate estimates when used for computing species richness within an ecosystem. Regardless of the approach utilized to estimate species richness, the estimates obtained are highly dependent on sample size, and smaller datasets typically result in the underestimation of species richness (14, 44, 47, 55).The use of a pyrosequencing-based approach (40) in 16S gene-based diversity surveys promises to overcome both of the above-mentioned problems associated with inadequate sampling. The large number of 16S rRNA gene sequences produced (hundreds of thousands) allows access to rare members of the community (25; J. M. Tiedje, presented at the 108th General Meeting of the American Society for Microbiology, Boston, MA, 2008), as well as a relatively more accurate estimation of species richness. However, with the introduction of this new technology, it is necessary to correlate the results obtained from newer pyrosequencing-based surveys to the extensive collection of longer, capillary sequence-generated 16S rRNA gene sequences that has been deposited in public databases during the last 2 decades. Several recent studies have examined the utility of pyrosequencing fragments in providing an accurate survey of overall community structure (36) and investigated the ability of various fragments spanning the 16S rRNA gene to accurately predict the phylogenetic affiliation of pyrosequencing-generated fragments at various taxonomic cutoffs (35, 54). As such, these admirable efforts gave useful insights into the advantages and limitations of the pyrosequencing approach in 16S-based community surveys, pinpointed specific regions that provide better phylogenetic resolution than other pyrosequencing-generated regions, and provided a quantitative assessment of binning accuracy at various empirical cutoffs.However, while issues regarding correlating phylogenies of shorter and longer fragments are actively being addressed, efforts to calibrate species richness data obtained from various pyrosequencing fragments at various taxonomic cutoffs to estimates obtained using longer 16S rRNA gene fragments are still lacking. It is unclear how pairwise distances and, hence, operational taxonomic unit (OTU) assignments and species richness estimates computed using various shorter fragments spanning various regions of the 16S rRNA gene will correlate to pairwise distances computed using the nearly complete 16S rRNA gene. Elucidating such differences between shorter and nearly complete fragments, as well as between shorter fragments representing different regions in the 16S rRNA gene, is absolutely necessary for accurate meta-analysis of species richness in previously published and future datasets constructed using various sequencing approaches.Here, we constructed, sequenced, and analyzed a 16S rRNA library of 1,132 clones generated from an undisturbed tallgrass prairie soil in central Oklahoma and compared the numbers of OTUs and species richness values obtained using the full-length data sets (with and without the application of the Lane mask filter that excludes hypervariable regions from the phylogenetic analysis) (32) and fragments simulating pyrosequencing output generated by clipping where known conserved bacterial primers are encountered in the 16S rRNA gene. The lengths of the chosen simulated-pyrosequencing fragments represent amplicons that have been generated using the original GS20 pyrosequencing platform (≈100 bp) (25, 44, 48), similar to those currently being generated using the GS FLX pyrosequencing platform (≈250 bp) (1, 20, 35) or amplicons produced using the anticipated increase in the new GS XLR pyrosequencing platform (>250 bp). We show that the choice of the pyrosequenced fragment could indeed impact the number of OTUs calculated at different taxonomic cutoffs, with some fragments underestimating and others overestimating such parameters compared to the results with longer, nearly complete 16S rRNA gene fragments. We also show that even more marked differences could be encountered when comparing two pyrosequencing fragments within the same molecule. Further, we established a regression analysis that explains the nature of the observed discrepancies using the proportions of the hypervariable, variable, and conserved bases within fragments.     

An efficient method for transformation of pre-androgenic,isolated <Emphasis Type="Italic">Brassica napus</Emphasis> microspores involving microprojectile bombardment and <Emphasis Type="Italic">Agrobacterium-</Emphasis>mediated transformation

The physical barrier imposed by the thick microspore wall constitutes an obstacle for an efficient Agrobacterium-mediated transformation of vacuolate microspores prior to androgenic induction and haploid embryogenic commitment. It is thus necessary to implement additional methods to overcome this drawback. In this study, we focused on the optimization of a protocol to allow for the exogenous DNA to enter the microspore in an efficient manner. We tested different options, based on microprojectile bombardment, to be applied prior to agroinfiltration. From them, the best results were obtained through co-transformation by microspore bombardment with DNA-coated microprojectile particles, followed by Agrobacterium tumefaciens infection. This method provides an efficient means to integrate extraneous DNA into rapeseed microspores prior to androgenesis induction.     

Radioprotective effects of hawthorn against genotoxicity induced by gamma irradiation in human blood lymphocytes

The radioprotective effect of hawthorn (Crataegus microphylla) fruit extract was investigated in cultured blood lymphocytes from human volunteers. Peripheral blood samples were collected from five human volunteers 10 min before and 1, 2 and 3 h after a single oral ingestion of 500 mg hawthorn powder extract. At each time point, the whole blood was exposed in vitro to 150 cGy of cobalt-60 gamma irradiation, and then the lymphocytes were cultured with mitogenic stimulation to determine the micronuclei in cytokinesis-blocked binucleated cell. The lymphocytes in the blood samples collected after extract ingestion exhibited a significant decrease in the incidence of binucleated cells containing micronuclei as compared to similarly irradiated lymphocytes collected prior to extract ingestion. The maximum decrease in the frequency of micronuclei-containing cells was observed at 1 h after ingestion of Hawthorn extract (on average a 44% decrease). These data suggest that it may be possible to use Hawthorn extracts in personnel exposed to radiation in order to protect lymphocytes from radiation effects.