DNA methylation and expression analyses reveal epialleles for the foliar disease resistance genes in peanut (Arachis hypogaea L.).

OBJECTIVE: Low DNA sequence polymorphism despite enormous phenotypic variations in peanut indicates the possible role of epigenetic variations. An attempt was made to analyze genome-wide DNA methylation pattern and its influence on gene expression across 11 diverse genotypes of peanut. RESULTS: The genotypes were subjected to bisulfite sequencing after 21 days of sowing (DAS). CHG regions showed the highest (30,537,376) DNA methylation followed by CpG (30,356,066) and CHH (15,993,361) across 11 genotypes. The B sub-genome exhibited higher DNA methylation sites (46,294,063) than the A sub-genome (30,415,166). Overall, the DNA methylation was more frequent in inter-genic regions than in the genic regions. The genes showing altered methylation and expression between the parent (TMV 2) and its EMS-derived mutant (TMV 2-NLM) were identified. Foliar disease resistant genotypes showed significant differential DNA methylation at 766 sites corresponding to 25 genes. Of them, two genes (Arahy.1XYC2X on chromosome 01 and Arahy.00Z2SH on chromosome 17) coding for senescence-associated protein showed differential expression with resistant genotypes recording higher fragments per kilobase of transcript per million mapped reads (FPKM) at their epialleles. Overall, the study indicated the variation in the DNA methylation pattern among the diverse genotypes of peanut and its influence of gene expression.

Study of structural and magnetic properties and heat induction of gadolinium-substituted manganese zinc ferrite nanoparticles for in vitro magnetic fluid hyperthermia.

This article outlines the synthesis of gadolinium (Gd)-doped manganese zinc ferrite magnetic nanoparticles (MNPs) as potential magnetic carriers for magnetic fluid hyperthermia (MFH). MNPs with high specific loss power (SLP; 146 W/g) have been developed and used for an in vitro hyperthermia study. The treatment of MFH is fruitful if there is an adequate number of MNPs in tumor cells with the highest SLP to rapidly generate heat while minimizing thermal injury to surrounding healthy tissue. X-ray diffraction patterns of the studied particles confirm the formation of a cubic spinel structure. Field emission scanning electron micrographs showed homogeneous distributions of particles with some agglomerates with a granular appearance. Transmission electron microscopy analysis showed the presence of agglomerated spherical particles at the surface. The substitution of Gd resulted in superparamagnetism at room temperature as confirmed by vibrating sample magnetometer analysis. The estimated saturation magnetization reduced from 48.6 to 28.2 emu/g with an increase in Gd concentration. However, the coercivity increased from 1093 Oe to 1597 Oe. Field cooled and zero field cooled measurements showed Curie temperatures from 315 to 326 K, as required for MFH applications. Cell viability measurements indicated that the MNPs are nontoxic to A549 cells for the studied concentrations of particle fraction and a contact time of up to 24 h. The interaction of the MNPs with A549 cells was highlighted from an image captured by an inverted microscope. In order to treat cancer in vivo, an in vitro hyperthermia study has initially been carried out with A549 cells.

Biological Activities and Identification of Bioactive Metabolite from Endophytic Aspergillus flavus L7 Isolated from Aegle marmelos.

Aegle marmelos, a well-known Indian plant with medicinal and religious importance, has been extensively used in Indian traditional medicine. The present study aimed to isolate, identify, and evaluate the biological activities of endophytic fungi from A. marmelos. One of the isolates, labeled as L7, was identified as Aspergillus flavus using morphology and ITS gene sequence. Total phenolic and flavonoid contents in the culture filtrate were found to be 65.77 mg GAE/ml and 158.33 mg quercetin/ml of crude extract, respectively. The extract showed excellent antimicrobial activity against common human bacterial and fungal pathogens. The test extract at 700 µg/ml, which notably reduced the concentration of DPPH-free radical as percent DPPH scavenging activity, was found to be the highest (64.53 %). The extract, at the concentration of 2 mg/ml, produced 70 % inhibition of hemolysis of RBCs compared to 78 % produced by standard drug (Ibuprofen). Chemical profiling of the fermented extract using TLC followed by UV and FTIR revealed the presence of flavonoids. The HPLC analysis confirmed the presence of bioflavonoid rutin in the extract. To the best of our knowledge, this is the first report on production of bioactive flavonoid by endophytic Aspergillus flavus obtained from A. marmelos and its pharmaceutical potential. In conclusion, the endophytic Aspergillus flavus obtained from the A. marmelos could be explored as an economic and potential natural resource with diverse pharmaceutical and biological activities.

Somatic cell count and alkaline phosphatase activity in milk for evaluation of mastitis in buffalo.

BACKGROUND AND AIM: Mastitis is a serious disease of dairy animals causing great economic losses due to a reduction in milk yield as well as lowering its nutritive value. The application of somatic cell count (SCC) and alkaline phosphatase activity in the milk for diagnosis of mastitis in buffalo is not well documented. Therefore, the present study was conducted to observe the SCC and alkaline phosphatase activity for evaluation of mastitis in buffalo. MATERIALS AND METHODS: Milk samples of forty apparently healthy lactating buffaloes were selected and categorized into five different groups viz. normal buffaloes, buffaloes with subclinical mastitis with CMT positive milk samples (+1 Grade), (+2 Grade), (+3 Grade), and buffaloes with clinical mastitis with 8 animals in each group. The milk samples were analyzed for SCC and alkaline phosphatase activity. RESULTS: The levels of SCC (×10(5) cells/ml) and alkaline phosphatase (U/L) in different groups were viz. normal (3.21±0.179, 16.48±1.432), subclinical mastitis with CMT positive milk samples with +1 Grade (4.21±0.138, 28.11±1.013), with +2 Grade (6.34±0.183, 34.50±1.034), with +3 Grade (7.96±0.213, 37.73±0.737) and buffaloes with clinical mastitis (10.21±0.220, 42.37±0.907) respectively, indicating an increasing trend in the values and the difference observed among various group was statistically significant. CONCLUSION: In conclusion, the results of the present study indicate that the concentration of milk SCC and alkaline phosphatase activity was higher in the milk of buffaloes with mastitis than in the milk of normal buffaloes.

Exploratory studies on incineration of carbaryl waste.

A hazardous waste stream, generated during the manufacture of carbaryl (1-naphthyl-n-methylcarbamate), an insecticide, was explored for assessing its suitability for incineration. The physico-chemical characteristics such as proximate analysis (moisture, volatile matter, fixed carbon and ash), ultimate analysis (carbon, hydrogen, nitrogen, sulphur and oxygen), calorific value and the heavy metal content of the waste indicated that carbaryl waste was suitable for incineration. The incineration experiments were conducted in a bench-scale (25 mm ID, 450 mm long) quartz reactor at various operating temperatures (800 to 1200 degrees C) at a fixed gas-phase residence time of two seconds. Results of the experiments showed that carbaryl waste could be effectively incinerated at a temperature of 1200 degrees C with a gas-phase residence time of two seconds. The destruction and removal efficiency (DE) at these operating conditions was > 99.99% for carbaryl, which was monitored as a principal organic hazardous compound (POHC). The ash generated at these operating conditions was subjected to the Toxicity Characteristic Leaching Procedure (TCLP) and was found to be non-toxic in nature.

An approach for modeling thermal destruction of hazardous wastes in circulating fluidized bed incinerator.

This paper presents a detailed computational fluid dynamics (CFD) based approach for modeling thermal destruction of hazardous wastes in a circulating fluidized bed (CFB) incinerator. The model is based on Eular - Lagrangian approach in which gas phase (continuous phase) is treated in a Eularian reference frame, whereas the waste particulate (dispersed phase) is treated in a Lagrangian reference frame. The reaction chemistry hasbeen modeled through a mixture fraction/ PDF approach. The conservation equations for mass, momentum, energy, mixture fraction and other closure equations have been solved using a general purpose CFD code FLUENT4.5. Afinite volume method on a structured grid has been used for solution of governing equations. The model provides detailed information on the hydrodynamics (gas velocity, particulate trajectories), gas composition (CO, CO2, O2) and temperature inside the riser. The model also allows different operating scenarios to be examined in an efficient manner.

Multi-level screening of a proposed hazardous waste treatment and disposal facility site--a case study.

A comprehensive environmental assessment (EA) is a pre-requisite before the site for developing a hazardous waste treatment and disposal facility (HWTDF) is selected. However, the resource limitations, especially for developing countries, often dictate that the detailed EA be restricted to those sites, which are constraint free and have low hazard potentials. Thus, a preliminary screening exercise for assessing the suitability of site for developing the HWTDF needs to be carried out to avoid huge costs involved in detailed EA. While screening a HWTDF site, various factors such as present land use, ecologically sensitive areas, geology and hydrogeology of the area, the quality and quantity of wastes, engineered safeguards, and the operational procedures that need to be adopted. are required to be addressed. In this paper, a multi-level screening criteria employing Remote Sensing, Constraint Mapping, Groundwater Pollution Potential Index (DRASTIC Index), and the Site Ranking was used to assess the suitability of a proposed site for the development of HWTDF. The study helped to identify various constraints at the proposed site and their significance on the development of the HWTDF.

Psocoptera spp. (book louse): a new major household allergen in Mumbai.

BACKGROUND: Exposure and sensitization to indoor allergens of house dust mites and insects is well established. However, there are no reports on the allergenicity of Psocoptera spp. (book louse). These insects are known to inhabit various terrestrial habitats such as tree bark (bark lice), old papers, books, stored food products, etc and gain entry into human dwellings through these modes. They have the potential to sensitize individuals with an occupational exposure to them. OBJECTIVE: Our objective is to use clinical and immunologic techniques to evaluate the extent of sensitization to Psocoptera in patients with nasobronchial allergy. We also have the objective to determine the extent of Psocoptera infestation in houses in Mumbai (Bombay). METHODS: An entomologic analysis of dust samples in houses in Mumbai was carried out and Psocoptera found in these samples was mass-cultured. Two hundred patients were checked for allergy to the Psocoptera extracts using skin test. Total IgE and specific IgE to Psocoptera in sera from Psocopotera-sensitive patients' sera were estimated by enzyme immunoassay. IgE-binding proteins of Psocoptera were determined by immunoblotting experiments. RESULTS: Among the 75 house dust samples from Mumbai analyzed for the presence of Psocoptera, 25% were found to be infested by Psocoptera. Strong skin sensitivity to Psocoptera was found in 20% of patients. Total IgE levels >150 IU/mL were found in these patients and high levels of specific IgE to Psocoptera were detected by ELISA. Immunoblot analysis revealed 67, 59, 43, and 27 kD proteins as major allergens of Psocoptera. CONCLUSIONS: Psocoptera is an important allergen in Mumbai as it causes IgE mediated responses in 20% of the allergic population studied.