DNA barcoding of tuberous Orchidoideae: a resource for identification of orchids used in Salep

Tubers of terrestrial orchids are harvested and traded from the eastern Mediterranean to the Caspian Sea for the traditional product Salep. Overexploitation of wild populations and increased middle‐class prosperity have escalated prices for Salep, causing overharvesting, depletion of native populations and providing an incentive to expand harvesting to untapped areas in Iran. Limited morphological distinctiveness among traded Salep tubers renders species identification impossible, making it difficult to establish which species are targeted and affected the most. In this study, a reference database of 490 nrITS, trnL‐F spacer and matK sequences of 133 taxa was used to identify 150 individual tubers from 31 batches purchased in 12 cities in Iran to assess species diversity in commerce. The sequence reference database consisted of 211 nrITS, 158 trnL‐F and 121 matK sequences, including 238 new sequences from collections made for this study. The markers enabled unambiguous species identification with tree‐based methods for nrITS in 67% of the tested tubers, 58% for trnL‐F and 59% for matK. Species in the genera Orchis (34%), Anacamptis (27%) and Dactylorhiza (19%) were the most common in Salep. Our study shows that all tuberous orchid species in this area are threatened by this trade, and further stresses the urgency of controlling illegal harvesting and cross‐border trade of Salep tubers.     

Chemical compositions of leaf extracts from Conocarpus erectus L. (Combretaceae) and their bioactivities against Tribolium castaneum Herbst (Coleoptera: Tenebrionidae)

Botanical insecticides have long been considered as alternatives to synthetic chemical insecticides in IPM programs. Effects of aqueous and ethanolic extracts obtained from buttonwood, Conocarpus erectus L. (Combretaceae), leaves on a major kind of stored product pests, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae), were evaluated under laboratory conditions. For this purpose, LC₅₀, repellency, antifeedant properties, and some biological effects (including body weight, immature developmental time and survival) of the insect were determined. The aqueous and ethanolic extracts were highly toxic to the larvae and adults. Calculated LC₅₀ values ranged between 2.6 and 193.4 (g/kg). Both extracts had repellent and antifeedant properties against the adults. The extracts adversely affected the larval and pupal weights, developmental time, and survival. Aqueous extracts were more effective only for the LC₅₀ values and only in females. All other measured parameters do not differ between the two extracts. The bioactive properties might be related to high amounts of alkaloid, phenol and tannin. Aqueous extract of the plant leaves may be a useful alternative for chemical insecticides.     

Plant growth promoting activity of an auxin and siderophore producing isolate of <Emphasis Type="Italic">Streptomyces</Emphasis> under saline soil conditions

A biocontrol Streptomyces isolate (C) was tested for its plant growth promoting qualities under saline conditions. Exposure to elevated osmotic strengths up to 300 mM NaCl increased dry weight and cfu/ml significantly. The isolate C produced indolyl-3-acetic acid (IAA) into the medium in the amount of 2.4 μg/ml. The amount of auxin increased after adding salt and reached to 4.7 μg/ml in 300 mM NaCl. Biosynthesis of siderophore was detectable and increased in presence of NaCl. Streptomyces isolate C showed good solubilization of tricalcium phosphate in culture medium with 92 mg/l. Solubilization decreased in presence of NaCl. Soil treatment with isolate C increased the growth and development of wheat plant in normal and saline conditions. In this treatment there were significant increases in germination rate, percentage and uniformity, shoot length and dry weight compared to the control. Applying the bacterial inocula increased the concentration of N, P, Fe and Mn in wheat shoots grown in normal and saline soil, but had non-significant effect on other micro and macronutrients concentrations. Results of this study show that Streptomyces isolate C has potential to be utilized as biofertilizer in saline soils.     

Molecular diversity of CTX prophage in Vibrio cholerae

Aims: The objective of this study was to investigate the molecular diversity of CTX genetic element within toxigenic Vibrio cholerae genomes and to determine the genetic diversity of V. cholerae population collected in a 6‐year period (2004–2009) in Iran. Methods and Results: The results of mismatch amplification mutation assay (MAMA)‐PCR and sequencing showed cytosine nucleotide in positions 203 and 115 in all 50 El Tor V. cholerae strains, which is the same as classical ctxB sequence. One strain yielded amplicons with both El Tor and classical biotype primers in MAMA‐PCR indicative of presence of two copies of CTX phages with different genotypes (rstR^ET ctxB^class and rstR^ET ctxB^ET) integrated within the genome of this isolate, which suggested the integration of two different CTX phages at different occasions or point mutation in one copy of CTX. Sequencing and PCR analysis indicated the presence of hybrid CTX genotype (rstR^ET ctx^class) in 70·6% of the isolates; however, only El Tor RS1 phage has been integrated in flanking to the CTX phages with different genotypes. Conclusions: Enterobacterial repetitive intergenic consensus‐PCR (ERIC‐PCR) and ribosomal gene spacer‐PCR (RS‐PCR) showed a relatively homogenous population in different years. Our findings indicate that sequence analysis of RS and ctxB regions has more discriminative power than restriction‐based methods. Significance and Impact of the Study: Investigating the molecular diversity of CTX prophage among V. cholerae strains helps to establish a new valuable database of genetic information about isolates, which is of great importance for epidemiologic studies in Iran and other countries encountering cholera epidemics.     

Myeloid deletion and therapeutic activation of AMPK do not alter atherosclerosis in male or female mice

The dysregulation of myeloid-derived cell metabolism can drive atherosclerosis. AMP-activated protein kinase (AMPK) controls various aspects of macrophage dynamics and lipid homeostasis, which are important during atherogenesis. Using LysM-Cre to drive the deletion of both the α1 and α2 catalytic subunits (MacKO), we aimed to clarify the role of myeloid-specific AMPK signaling in male and female mice made acutely atherosclerotic by injection of AAV vector encoding a gain-of-function mutant PCSK9 (PCSK9-AAV) and WD feeding. After 6 weeks of WD feeding, mice received a daily injection of either the AMPK activator A-769662 or a vehicle control for an additional 6 weeks. Following this (12 weeks total), we assessed myeloid cell populations and differences between genotype or sex were not observed. Similarly, aortic sinus plaque size, lipid staining, and necrotic area did not differ in male and female MacKO mice compared with their littermate floxed controls. Moreover, therapeutic intervention with A-769662 showed no treatment effect. There were also no observable differences in the amount of circulating total cholesterol or triglyceride, and only minor differences in the levels of inflammatory cytokines between groups. Finally, CD68+ area and markers of autophagy showed no effect of either lacking AMPK signaling or AMPK activation. Our data suggest that while defined roles for each catalytic AMPK subunit have been identified, complete deletion of myeloid AMPK signaling does not significantly impact atherosclerosis. Additionally, these findings suggest that intervention with the first-generation AMPK activator A-769662 is not able to stem the progression of atherosclerosis.     

Anti Diabetic effect of CL 316,243 (A β3-Adrenergic Agonist) by Down Regulation of Tumour Necrosis Factor (TNF-α) Expression

Objective

Obesity is a risk factor for the development of insulin resistance and is one of the most important contributors to the pathogenesis of type2 diabetes, which acts mainly through the secretion of adipokines such as TNF-α that may influence insulin sensitivity. TNF-α affects many aspects of adipocyte function, such as adipocyte development and lipid metabolism.

Material and Methods

We demonstrated that there is a correlation between the expressions of TNF-α in retroperitoneal WAT and insulin-resistance in 8 genetically obese fa/fa rats. Treatment of animals with CL 316,243, a β3-adrenergic agonist, showed an improvement of insulin-resistance that was linked with the suppression of TNF-α mRNA expression in WAT.

Results

These results confirm the association between TNF-α expression and the insulin-resistant condition in rats. Our finding indicates that the hyperglycaemia and hyperinsulinemia induced by insulin-resistance correlated positively with the expression of TNF-α mRNA in an abdominal WAT depot.

Conclusion

We conclude that CL 316,243 possesses both anti-diabetic effects and anti-obesity effects in rodents.     

A comparison of symmetry and flatness measurements in small electron fields by different dosimeters in electron beam radiotherapy

BackgroundSymmetry and flatness are two quantities which should be evaluated in the commissioning and quality control of an electron beam in electron beam radiotherapy. The aim of this study is to compare symmetry and flatness obtained using three different dosimeters for various small and large fields in electron beam radiotherapy with linac.Materials and methodsBeam profile measurements were performed in a PTW water phantom for 10, 15 and 18 MeV electron beams of an Elekta Precise linac for small and large beams (1.5 × 1.5 cm² to 20 × 20 cm² field sizes). A Diode E detector and Semiflex-3D and Advanced Markus ionization chambers were used for dosimetry.ResultsBased on the obtained results, there are minor differences between the responses from different dosimeters (Diode E detector and Semiflex-3D and Advanced Markus ionization chambers) in measurement of symmetry and flatness for the electron beams. The symmetry and flatness values increase with increasing field size and electron beam energy for small and large field sizes, while the increases are minor in some cases.ConclusionsThe results indicate that the differences between the symmetry and flatness values obtained from the three dosimeter types are not practically important.