Strain-Specific Behavior of Mycobacterium tuberculosis in Interruption of Autophagy Pathway in Human Alveolar Type II Epithelial A549 Cells

Background:Autophagy induction has been shown to differ in magnitude depending on the mycobacterial species. However, few studies have investigated the specific autophagic capacity of different Mtb strains in ATs. This study aimed to elucidate the host autophagic response to different Mtb strains in ATs responsible for TB in the capital of Iran, Tehran. Methods:A549 cells were infected with three different Mtb clinical isolates (Beijing, NEW1, and CAS1/Delhi) and the reference strain H37Rv. Following RNA extraction, the expression of eight ATG genes, four mycobacterial genes, and three miRNAs was evaluated using quantitative RT-PCR. Results:The results revealed that all four strains influenced the autophagy pathway in various ways at different magnitudes. The Beijing and H37Rv strains could inhibit autophagosome formation, whereas the CAS and NEW1 strains induced autophagosome formation. The expression of genes involved in the fusion of autophagosomes to lysosomes (LAMP1) indicated that all the studied strains impaired the autophagolysosomal fusion; this result is not unexpected as Mtb can block the autophagolysomal fusion. In addition, the Beijing and H37RV strains prevented the formation of autophagic vacuoles, besides mycobacterial targeting of lysosomes and protease activity.Conclusion:This preliminary study improved our understanding of how Mtb manages to overcome the host immune system, such as autophagy, and evaluated the genes used by specific strains during this process. Further studies with a large number of Mtb strains, encompassing the other main Mtb lineages, are inevitable.Key Words: A549 cells, Autophagy, MicroRNA, Mycobacterium tuberculosis     

Regulatory Effect of let-7f Transfection in Non-Small Cell Lung Cancer on its Candidate Target Genes

Background:Let-7f has essential impacts on biological processes; however, its biological and molecular functions in lung cancer pathogenesis have yet been remained unclear. We aimed to investigate the expression level of let-7f and its candidate target genes both in lung cancer tissues and A549 cell line. Methods:Bioinformatics databases were first used to select candidate target genes of let-7f. Then the relative gene and protein expressions of let-7f and its target genes, including HMGA2, ARID3B, SMARCAD1, and FZD3, were measured in lung tissues of Non-Small Cell Lung Cancer (NSCLC) patients and A549 cell line using quantitative real-time PCR and Western blotting. The electroporation method was used to transfect A549 cells with let-7f mimic and microRNA inhibitor. The impact of let-7f transfection on the viability of A549 cells was assessed using MTT assay. The expression data of studied genes were analyzed statistically Results:Results indicated significant downregulated expression level of let-7f-5p (p = 0.0013) and upregulated level of the HMGA2 and FZD3 in NSCLC cases (p < 0.05). In A549 cells, after transfection with let-7f mimic, the expression of both mRNA and protein levels of HMGA2, ARID3B, SMARCAD1, and FZD3 decreased. Also, the overexpression of let-7f significantly inhibited the A549 cell proliferation and viability (p = 0.017). Conclusion:Our findings exhibited the high value of let-7f and HMGA2 as biomarkers for NSCLC. The let-7f, as a major tumor suppressor regulatory factor via direct targeting genes (e.g. HMGA2), inhibits lung cancer cell viability and proliferation and could serve as a marker for the early diagnostic of NSCLC.Key Words: MicroRNAs, Biomarkers, Carcinoma, Non-Small-Cell Lung     

Influence of fumigation with high concentrations of ozone gas on postharvest gray mold and fungicide residues on table grapes

To control postharvest decay, table grapes are commercially fumigated with sulfur dioxide. We evaluated ozone (O₃) fumigation with up to 10,000 μL L^?1 of ozone for up to 2 h to control postharvest gray mold of table grapes caused by Botrytis cinerea. Fumigation for 1 h with 2500 or 5000 μL L^?1 of ozone were equal in effectiveness. Both treatments reduced postharvest gray mold among inoculated ‘Thompson Seedless’ grapes by approximately 50% when the grapes were examined after storage for 7 d at 15 °C following fumigation. In a similar experiment, ‘Redglobe’ grapes were stored for 28 d at 0.5 °C following fumigation for 1 h with 2500 or 5000 μL L^?1 of ozone. Both treatments were equal in effectiveness, but inferior to fumigation with 10,000 μL L^?1. Ozone was effective when grapes were inoculated and incubated at 15 °C up to 24 h before fumigation. The cluster rachis sustained minor injuries in some tests, but berries were never harmed. Ozone was applied in three combinations of time and ozone concentration (10,000 μL L^?1 for 30 min, 5000 μL L^?1 for 1 h, and 2500 μL L^?1 for 2 h) where each had a constant concentration × time product (c × t) of 5000 μL L^?1 × h. The effectiveness of each combination was similar. The incidence of gray mold was reduced by approximately 50% among naturally inoculated, organically grown ‘Autumn Seedless’ and ‘Black Seedless’ table grapes, and by 65% among ‘Redglobe’ table grapes, when they were fumigated with 5000 μL L^?1 ozone for 60 min in a commercial ozone chamber and stored for 6 weeks at 0.5 °C. Residues of fenhexamid, cyprodinil, pyrimethanil, and pyraclostrobin were reduced by 68.5, 75.4, 83.7, and 100.0%, respectively, after a single fumigation of table grapes with 10,000 μL L^?1 ozone for 1 h. Residues of iprodione and boscalid were not significantly reduced. Ozone is unlikely to replace sulfur dioxide treatments in conventional grape production unless its efficacy is improved, but it could be an acceptable technology to use with grapes marketed under “organic” classification, where the use of SO₂ is prohibited, or if SO₂ use were to be discontinued.     

Selective genotyping to identify late blight resistance genes in an accession of the tomato wild species <Emphasis Type="Italic">Solanum pimpinellifolium</Emphasis>

Late blight (LB), caused by the oomycete Phytophtohra infestans, is one of the most destructive diseases of tomato (Solanum lycopersicum) and other Solanaceae species. Current disease control and prevention strategies are not sufficient to control the disease in tomato. Recent germplasm screening experiments led to the identification of a new source of resistance (PI 270443) in the tomato wild species S. pimpinellifolium. This study was conducted to identify genomic regions associated with LB resistance in this accession. A large F₂ population (n = 986) derived from a cross between PI 270443 and a LB-susceptible tomato breeding line (NCEBR-2) was screened for LB resistance using a highly aggressive isolate of P. infestans. Twenty-five of the most resistant and 29 of the most susceptible, but surviving F₂ individuals were identified based on disease evaluations conducted in the F₂ and F₃ progeny populations. The selected individuals were genotyped with 153 DNA markers located across the 12 tomato chromosomes. A selective genotyping approach led to the identification of two genomic regions on tomato chromosomes 1 and 10 associated with LB resistance in PI 270443. Identification of two genomic regions associated with resistance was consistent with a previous estimate of the number of LB resistance genes in this accession. Research is currently underway to fine map the two resistance genes and incorporate them into new tomato breeding lines and hybrid cultivars.     

Interaction of water and nitrogen on maize grown for silage

Water scarcity and environmental pollution due to excessive nitrogen (N) applications are important environmental concerns. The Varamin region, which is located in the central part of Iran, is one of the locations where farmers apply 250-350 kg N ha⁻¹ for silage maize without any concerns with respect to the available water for irrigation. The objective of this study was to quantify the response of the silage maize (Zea mays L.) to variable irrigation and N fertilizer applications under arid and semi-arid conditions and to determine the optimum amount of N fertilizer as a function of irrigation. The maize Hybrid 704 single-cross was planted on 3 August 2003 and on 25 June 2004. The experimental treatments consisted of three N rates (0, 150, and 200 kg N ha⁻¹) and four levels of irrigation, including two deficit irrigation levels 0.70 SWD (soil water depletion) and 0.85 SWD, a full-irrigation level (1.0 SWD) and an over-irrigation level (1.13 SWD). Twelve treatments were arranged in a strip-plot design in a randomized complete block with three replicates. Gravimetric soil samples were collected in 2003 and a neutron probe was used in 2004 to measure soil water content. Leaf area index, total aboveground biomass (TB), plant height, stem diameter, and leaf, stem, and ear dry weight were measured during the growing seasons and at final harvest. Total aboveground biomass was affected by irrigation (P < 0.0001) during both years and was also affected by N fertilizer in 2003 (P = 0.0001) and 2004 (P < 0.0001). However, there was no irrigation and N fertilizer interaction for both years (P > 0.5). Total aboveground biomass and biomass of the crop components increased as a function of the amount of water and N applied. For each of the irrigation levels, there was an associated optimum amount of N, which increased as the amount of irrigation water that was applied increased. Among the four irrigation levels that were studied, 0.85 SWD was the optimum level of irrigation for the conditions at the experimental site. The results also indicated that an increase in N applications is not a good strategy to compensate for a decrease of TB under drought stress conditions. We concluded that the effect of N fertilizer on TB depends on the availability of water in the soil, and that the amount of N fertilizer applied should be decreased under drought stress conditions. Further research will combine these results with a crop simulation model to help optimize nitrogen and water management for silage maize.     

GROWTH AND FLOWERING OF TUBEROSE AS AFFECTED BY ADDING NATURAL ZEOLITE TO THE CULTURE MEDIUM

This research was conducted to investigate the effect of natural zeolite on vegetative growth and flower characteristics of tuberose (Polianthes tuberosa L.). The experiment was performed as a completely randomized design with seven treatments (5, 10, 15, 20, 25 and 30 g zeolite/kg soil mixture along with control) and four replicates. Results indicate that zeolite increased the height of flowering stem, diameter of flower, leaf area and chlorophyll content, but decreased the diameter of flowering stem, dry weight of shoots, and roots and bulblets. Overall, zeolite added to the soil mixture had beneficial effects on only some of the parameters of tuberose growth and development.     

Picric Acid Degradation in Sediments from the Louisiana Army Ammunition Plant

Picric acid is an explosive historically produced and disposed at the Louisiana Army Ammunition Plant (LAAP) in northern Louisiana. The potential for natural degradation of picric acid was investigated by creating picric-acid slurries with four LAAP sediments of variable composition and monitoring for up to 98 days. The concentrations of picric acid decreased rapidly in all slurries during the first day, attributed to adsorption, followed by slower decreases in some samples due to degradation. Degradation in unsterilized slurries was nearly complete within 80 days for two of the four sediments. Increases in nitrite and nitrate concentration over time were proportional to the loss of picric acid and indicate that at least two of the three nitrite groups were removed from the picric acid molecule. The absence of significant concentrations of compounds with a mass greater than 100 amu in the final solutions suggests that all three nitrite groups were removed. No correlation was found between the degree of degradation and grain size, clay content, organic content, carbonate content, or a suite of element concentrations in the sediment. Degradation in sterilized samples was minimal for all sediment slurries, indicating microbial activity as the primary mechanism of degradation.     

Minimizing Ammonia Volatilization from Urea in Waterlogged Condition Using Chicken Litter Biochar

Application of urea in lowland rice fields leads to ammonia (NH₃) volatilization and environmental pollution, and diminishes nitrogen recovery by rice (Oryza sativa L.). Amending urea with biochar could reduce NH₃ loss from urea as well as improve chemical properties of acid soils. An incubation study was conducted using a closed-dynamic air flow system to determine NH₃ volatilization from urea and chemical properties of an acid soil (Typic Paleudults). The soil was mixed with three rates of chicken litter biochar (20, 40, and 60 g pot^?1) and 1.31 g urea. Mixing an acid soil with biochar (60 g pot^?1) in waterlogged to stimulate conditions in paddy condition significantly reduced NH₃ loss and total titratable acidity. Biochar application also increased soil pH, total nitrogen, available nitrate, organic matter, total organic carbon, total carbon, available phosphorus, and exchangeable cations. Thus, chicken litter biochar can be used to reduce urea-N loss and ameliorate chemical properties of acid soils. This aspect is being embarked on in our on-going field experiments.