Redirection of tryptophan metabolism in tobacco by ectopic expression of an Arabidopsis indolic glucosinolate biosynthetic gene

Indole-3-acetaldoxime (IAOx) is a branch point compound of tryptophan (Trp) metabolism in glucosinolate-producing species such as Arabidopsis, serving as a precursor to indole-glucosinolates (IGs), the defense compound camalexin, indole-3-acetonitrile (IAN) and indole-3-acetic acid (IAA). We synthesized [²H₅] and [¹³C₁₀¹⁵N₂]IAOx and [¹³C₆], [²H₅] and [2′,2′-²H₂]IAN in order to quantify endogenous IAOx and IAN in Arabidopsis and tobacco, a non-IG producing species. We found that side chain-labeled [2′,2′-²H₂]IAN overestimated the amount of IAN by 2-fold compared to when [²H₅]IAN was used as internal standard, presumably due to protium-deuterium exchange within the internal standard during extraction of plant tissue. We also determined that [¹³C₁]IAN underestimated the amount of IAN when the ratio of [¹³C₁]IAN standard to endogenous IAN was greater than five to one, whereas either [²H₅]IAN or [¹³C₆]IAN showed a linear relationship with endogenous IAN over a broader range of concentrations. Transgenic tobacco vector control lines did not have detectable levels of IAOx or IAN (limit of detection ∼ 100 pg/g fr. wt), while lines expressing either the IAOx-producing CYP79B2 or CYP79B3 genes from Arabidopsis under CaMV 35S promoter control accumulated IAOx in the range of 1-9 μg/g fr. wt. IAN levels in these lines ranged from 0.6 to 6.7 μg/g fr. wt, and IAA levels were ∼9-14-fold above levels in control lines. An Arabidopsis line expressing the same CYP79B2 overexpression construct accumulated IAOx in two of three lines measured (∼200 and 400 ng/g fr. wt) and accumulated IAN in all three lines. IAN is proposed to be a metabolite of IAOx or an enzymatic breakdown product of IGs induced upon tissue damage. Since tobacco does not produce detectable IGs, the tobacco data are consistent with IAN being a metabolite of IAOx. IAOx and IAN were also examined in the Arabidopsis activation tagged yucca mutant, and no accumulation of IAOx was found above the limits of detection but accumulation of IAN (3-fold above wt) occurred. The latter was surprising in light of recent reports that rule out IAOx and IAN as intermediates in YUCCA-mediated IAA synthesis.     

Involvement of PI3K/Akt pathway in the protective effect of hesperidin against a chemically induced liver cancer in rats

Hesperidin is a flavanone glycoside that is found in the Citrus species and showed antioxidant, hepatoprotective as well as anticancer activity. This study investigated the effect of hesperidin on the PI3K/Akt pathway as a possible mechanism for its protective effect against diethylnitrosamine (DEN)‐induced hepatocellular carcinoma (HCC). Adult Wistar rats were divided into Control group (received drug vehicle); DEN group (received 100 mg/L of DEN solution for 8 weeks), and hesperidin + DEN group (received 200 mg/kg body weight of hesperidin/day orally for 16 weeks + DEN solution as DEN group). Our findings showed that the administration of hesperidin significantly decreased the elevation in liver function enzymes, serum AFP level, and oxidative stress markers. Moreover, hesperidin administration suppressed DEN‐induced upregulation of PI3K, Akt, CDK‐2 protein expression, and preserved the integrity of the liver tissues from HCC formation. In conclusion, the hepatoprotective activity of hesperidin is mediated via its antioxidation and downregulation of the PI3K/Akt pathway.     

Nine eriophyoid mite species from Iran (Acari, Eriophyidae)

Nine eriophyoid mites, including two new species and five new records, from Iran are described and illustrated. They are Aceria acroptiloni Shevchenko & Kacalev, 1974, rec. n. on Rhaponticum repens (L.) Hidalgo (Asteraceae); Aceria anthocoptes (Nalepa, 1892), rec. n. on Cirsium arvense (L.) Scop. (Asteraceae); Aceria lactucae (Canestrini, 1893), rec. n. on Lactuca virosa L. (Asteraceae); Aceria pulicarissp. n. on Pulicaria gnaphalodes (Vent.) Boiss. (Asteraceae); Aceria tosichella Keifer, 1969 on Setaria viridis (L.) Beauv. (Poaceae); Eriophyes rotundae Mohanasundaram, 1983 on Cyperus rotundus L. (Cyperaceae); Aculops maroccensis Keifer, 1972, rec. n. on Mentha piperita L. (Lamiaceae); Aculus medicagersp. n. on Medicago sativa L. (Leguminosae); Tetra lycopersici Xue & Hong, 2005, rec. n. on Solanum nigrum L. (Solanaceae).     

Serum testosterone in females exposed to natural sour gas with respect to polymorphisms of <Emphasis Type="Italic">XRCC1</Emphasis>, <Emphasis Type="Italic">GSTM1</Emphasis>, and <Emphasis Type="Italic">GSTT1</Emphasis>

The present study was done to determine the modulation effect(s) of polymorphisms of XRCC1, GSTM1, and GSTT1 on concentration of serum testosterone in females exposed to natural sour gas. Also we examine whether chronic exposure to natural gas containing sulfur compounds act as natural selection force on XRCC1 polymorphisms. The present study was performed on 68 healthy unrelated female students living in polluted areas of MIS. Also for investigating the effect of natural selection on XRCC1 polymorphism, a study was performed on two groups of healthy individuals of MIS citizens. The first and second groups including 94 (age range 30–85 years) and 187 individuals (age range 5–20 years), respectively. First and second groups were born and were not born in contaminated areas of the MIS, respectively. There was no significant difference between genotypes of XRCC1 for concentration of serum testosterone. Although GSTT1-null genotype had higher level of serum testosterone in comparison with the present genotype (t = 2.392, df = 66, P = 0.023), a borderline difference between genotypes of GSTM1 for serum testosterone was observed (t = 1.928, df = 66, P = 0.058). Analysis of variance revealed significant difference between combination genotypes of GSTM1 and GSTT1 for serum testosterone (F = 4.167; df = 3, 64; P = 0.009). The Duncan post hoc test indicated that the combination genotype of “present GSTM1/null GSTT1” had significant higher level of testosterone. There is no evidence that XRCC1 polymorphisms have advantage/disadvantage when population exposed to natural sour gas. The polymorphisms of GSTM1 and GSTT1 modulate serum testosterone concentration in young females exposed to natural sour gas.     

hiPSC-derived hepatocytes closely mimic the lipid profile of primary hepatocytes: A future personalised cell model for studying the lipid metabolism of the liver

Hepatocyte-like cells (HLCs) differentiated from human-induced pluripotent stem cells offer an alternative platform to primary human hepatocytes (PHHs) for studying the lipid metabolism of the liver. However, despite their great potential, the lipid profile of HLCs has not yet been characterized. Here, we comprehensively studied the lipid profile and fatty acid (FA) metabolism of HLCs and compared them with the current standard hepatocyte models: HepG2 cells and PHHs. We differentiated HLCs by five commonly used methods from three cell lines and thoroughly characterized them by gene and protein expression. HLCs generated by each method were assessed for their functionality and the ability to synthesize, elongate, and desaturate FAs. In addition, lipid and FA profiles of HLCs were investigated by both mass spectrometry and gas chromatography and then compared with the profiles of PHHs and HepG2 cells. HLCs resembled PHHs by expressing hepatic markers: secreting albumin, lipoprotein particles, and urea, and demonstrating similarities in their lipid and FA profile. Unlike HepG2 cells, HLCs contained low levels of lysophospholipids similar to the content of PHHs. Furthermore, HLCs were able to efficiently use the exogenous FAs available in their medium and simultaneously modify simple lipids into more complex ones to fulfill their needs. In addition, we propose that increasing the polyunsaturated FA supply of the culture medium may positively affect the lipid profile and functionality of HLCs. In conclusion, our data showed that HLCs provide a functional and relevant model to investigate human lipid homeostasis at both molecular and cellular levels.     

Th17 immune response to adipose tissue-derived mesenchymal stromal cells

Adipose tissue-derived mesenchymal stromal cells (ASCs) hold the promise of achieving successful immunotherapeutic results due to their ability to regulate different T-cell fate. ASCs also show significant adaptability to environmental stresses by modulating their immunologic profile. Cell-based therapy for inflammatory diseases requires a detailed understanding of the molecular relation between ASCs and Th17 lymphocytes taking into account the influence of inflammation and cell ratio on such interaction. Accordingly, a dose-dependent increase in Th17 generation was only observed in high MSC:T-cell ratio with no significant impact of inflammatory priming. IL-23 receptor (IL-23R) expression by T cells was not modulated by ASCs when compared to levels in activated T cells, while ROR-γt expression was significantly increased reaching a maximum in high (1:5) unprimed ASC:T-cell ratio. Finally, multiplex immunoassay showed substantial changes in the secretory profile of 15 cytokines involved in the Th17 immune response (IL-1β, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-22, IL-21, IL-23, IL-25, IL-31, IL-33, IFN-γ, sCD40, and TNF-α), which was modulated by both cell ratio and inflammatory priming. These findings suggest that Th17 lymphocyte pathway is significantly modulated by ASCs that may lead to immunological changes. Therefore, future ASC-based immunotherapy should take into account the complex and detailed molecular interactions that depend on several factors including inflammatory priming and cell ratio.     

Therapeutic Efficacy of Artemether-Lumefantrine (Coartem?) in Treating Uncomplicated P. falciparum Malaria in Metehara,Eastern Ethiopia: Regulatory Clinical Study

BackgroundAs per the WHO recommendation, the development of resistance by P. falciparum to most artemisinin combination therapies (ACTs) triggered the need for routine monitoring of the efficacy of the drugs every two years in all malaria endemic countries. Hence, this study was carried out to assess the therapeutic efficacy of Artemether-Lumefantrine (Coartem®) in treating the uncomplicated falciparum malaria, after 9 years of its introduction in the Metehara, Eastern Ethiopia.MethodThis is part of the therapeutic efficacy studies by the Federal Ministry of Health Ethiopia, which were conducted in regionally representative sentinel sites in the country from October 2014 to January 2015. Based on the study criteria set by WHO, febrile and malaria suspected outpatients in the health center were consecutively recruited to study. A standard six-dose regimen of AL was administered over three days and followed up for measuring therapeutic responses over 28 days. Data entry and analysis was done by using the WHO designed Excel spreadsheet and SPSS version 20 for Windows. Statistical significant was considered for P-value less than 0.05.ResultOf the 91 patients enrolled, the day-28 analysis showed 83 adequate clinical and parasitological responses (ACPRs). Per protocol analysis, PCR-uncorrected & corrected cure rates of Coartem® among the study participants were 97.6% (95%CI: 93.6–99.5) and 98.8% (CI: 93.5–100%), respectively. No parasite detected on day 3 and onwards. Fever clearance was above 91% on day-3. Mean hemoglobin was significantly increased (P<0.000) from 12.39 g/dl at day 0 to 13.45 g/dl on day 28. No serious adverse drug reactions were observed among the study participants.ConclusionThis study showed high efficacy of AL in the study area, which suggests the continuation of AL as first line drug for the treatment of uncomplicated P. falciparum malaria in the study area. This study recommends further studies on drug toxicity, particularly on repeated cough and oral ulceration.     

Thermoluminescent properties of some phosphor glasses based on aluminum oxide

The transparent thermoluminescent aluminum oxide‐based glass of 15Al₂O₃–35P₂O₅–25CaO–25Na₂CO₃, abbreviated as APCN (all in mol%) doped with different concentrations of SiO₂ from 0.0–500 ppm was prepared using a conventional melt‐quenching technique. The TL sensitivities of the prepared glasses were investigated at 3 Gy γ‐dose using a ⁶⁰Co source and measured at a heating rate 10 C/sec. The highest TL intensity of the material doped with SiO₂ was found at a concentration of 500 ppm (APCNSi₅). Deconvolution of the glow curve from APCNSi₅ resulted in four peaks at about 161, 194, 237 and 293 C with a Figure Of Merit (FOM) of 1.28%. The APCNSi₅ specimen had the best dosimetric properties when compared with the other samples. Reproducibility, repeatability, dose–response curve and fading effect were checked for peak 3, which appeared at about 237 C. The results displayed that the APCNSi₅ glass system was a low‐Z material (Z_eff ≈ 10), and had good reproducibility and good repeatability. Peak 3 showed good linearity over a dose range up to 20 Gy (R² = 0.999) and sublinearity behaviour was found. The signal from APCNSi₅ faded by about 11% after 2 days post irradiation, therefore it showed almost no significant loss. Such properties make the newly prepared glasses suitable for and highly recommended for use in γ‐dosimeters. Copyright © 2016 John Wiley & Sons, Ltd.