Introgression of blast and bacterial blight disease resistance genes in a rice genotype ADT43 through marker assisted back cross breeding.

Bacterial Leaf Blight (Xanthomonas oryzae pv. oryzae) and blast (Magnaporthe oryzae) are the major biotic stresses around the rice-growing zones of the world. The development of resistant varieties through Marker Assisted Backcross Breeding is the utmost economical and eco-friendly method for achieving stable yield. Amongst the resistance genes recognized, Xa21 and Pi54 possess broad-spectrum resistance to many Xoo and blast strains around the world. In the present study, we have effectively introgressed a Bacterial Blight resistance gene (Xa21) and a blast resistance gene (Pi54) into susceptible variety ADT43 from RP-Bio-Patho-2 coupled with phenotypic selection for agronomic, cooking quality and grain traits through MABC. MABC was sustained till BC2F2 generation with specific markers pTA248 for Xa21 and Pi54MAS for Pi54 resistance genes. A set of SSR markers for parental polymorphism were utilized for maximum regaining of recurrent parent genome in each backcrossing. "Positive plants" from BC2F1 were selfed to generate BC2F2 and the homozygous lines for bacterial leaf blight and blast resistance genes were identified for further assessment.

Comparison of trapezius squeeze test and jaw thrust as clinical indicators for laryngeal mask airway insertion in spontaneously breathing children.

BACKGROUND AND AIMS: It is not known whether trapezius squeeze test (TPZ) is a better clinical test than jaw thrust (JT) to assess laryngeal mask airway (LMA) insertion conditions in children under sevoflurane anesthesia. MATERIAL AND METHODS: After the Institutional Ethics Committee approval and written informed parental consent, 124 American Society of Anesthesiologists I and II children of 2-8 years of age undergoing minor surgical procedures were randomized into TPZ and JT groups. The children were induced with 8% sevoflurane in oxygen at a fresh gas flow of 4 L/min. TPZ or JT was performed after 1 min of start of sevoflurane and then every 20 s till the test was negative, when end-tidal (ET) sevoflurane concentration was noted. Classic LMA of requisite size was inserted by a blinded anesthetist and conditions at the insertion of LMA, insertion time, and the number of attempts of LMA insertion were recorded. RESULTS: The mean LMA insertion time was significantly longer (P < 0.001) for TPZ (145 ± 28.7 sec) compared to JT group (111.8 ± 31.0 sec). ET sevoflurane concentration at the time of LMA insertion was comparable in the two groups. LMA insertion conditions were similar in the two groups. There was no difference between the two groups regarding total number of attempts of LMA insertion. Heart rate (HR) decreased in both groups after LMA insertion (P < 0.001) but TPZ group had significantly lower HR compared with the JT group up to 5 min after LMA insertion (P = 0.03). CONCLUSION: Both JT and TPZ are equivalent clinical indicators in predicting the optimal conditions of LMA insertion in spontaneously breathing children; however, it takes a longer time to achieve a negative TPZ squeeze test.

Age-related sperm DNA methylation changes are transmitted to offspring and associated with abnormal behavior and dysregulated gene expression.

Advanced paternal age (APA) has been shown to be a significant risk factor in the offspring for neurodevelopmental psychiatric disorders, such as schizophrenia and autism spectrum disorders. During aging, de novo mutations accumulate in the male germline and are frequently transmitted to the offspring with deleterious effects. In addition, DNA methylation during spermatogenesis is an active process, which is susceptible to errors that can be propagated to subsequent generations. Here we test the hypothesis that the integrity of germline DNA methylation is compromised during the aging process. A genome-wide DNA methylation screen comparing sperm from young and old mice revealed a significant loss of methylation in the older mice in regions associated with transcriptional regulation. The offspring of older fathers had reduced exploratory and startle behaviors and exhibited similar brain DNA methylation abnormalities as observed in the paternal sperm. Offspring from old fathers also had transcriptional dysregulation of developmental genes implicated in autism and schizophrenia. Our findings demonstrate that DNA methylation abnormalities arising in the sperm of old fathers are a plausible mechanism to explain some of the risks that APA poses to resulting offspring.

A maize α-zein promoter drives an endosperm-specific expression of transgene in rice.

An alpha-zein promoter isolated from maize containing P-box, E motif sequence TGTAAAGT, opaque-2 box and TATA box was studied for its tissue-specific expression in rice. A 1,098 bp promoter region of alpha-zein gene, fused to the upstream of gusA reporter gene was used for transforming rice immature embryos (ASD 16 or IR 64) via the particle bombardment-mediated method. PCR analysis of putative transformants demonstrated the presence of transgenes (the zein promoter, gusA and hpt). Nineteen out of 37 and two out of five events generated from ASD 16 and IR 64 were found to be GUS-positive. A histological staining analysis performed on sections of mature T1 seeds revealed that the GUS expression was limited to the endosperm and not to the pericarp or the endothelial region. GUS expression was observed only in the following seed development stages : milky (14-15 DAF), soft dough (17-18 DAF), hard dough (20-23 DAF), and mature stages (28-30 DAF) of zein-gusA transformed (T0) plants. On the contrary a constitutive expression of GUS was evident in CaMV35S-gusA plants. PCR and Southern blotting analyses on T1 plants demonstrated a stable integration and inheritance of transgene in the subsequent T1 generation. GUS assay on T2 seeds revealed that the expression of gusA gene driven by alpha-zein promoter was stable and tissue-specific over two generations. Results suggest that this alpha-zein promoter could serve as an alternative promoter to drive endosperm-specific expression of transgenes in rice and other cereal transformation experiments.

Profiling of Groundnut bud necrosis orthotospovirus-responsive microRNA and their targets in tomato based on deep sequencing.

Tomato, an extensively cultivated vegetable crop produces miRNAs in response to infection with Groundnut bud necrosis orthotospovirus, a viral pathogen causing significant economic losses. High-throughput miRNA sequencing was performed on tomato leaves inoculated with GBNV and mock-inoculated leaves as controls. Analysis revealed 73 known miRNAs belonging to 24 miRNA families, with variable expression levels. Interestingly, 39 miRNAs were upregulated, and 34 were downregulated in response to GBNV infection. Stem-loop quantitative reverse transcription PCR validated the differential expression of selected miRNAs. Additionally, 30 miRNA encoded proteins were identified to be involved in disease resistance and susceptibility. The miRNA-target interactions were found to play significant roles in cellular and metabolic activities, as well as modulating signaling pathways during the plant-virus interaction. The findings shed light on the intricate regulatory network of miRNAs in tomato response to viral infection and may contribute to developing strategies for improving crop protection against viral diseases.

Chest wall radiotherapy with volumetric modulated arcs and the potential role of flattening filter free photon beams.

PURPOSE: The goal of the work was to assess the role of RapidArc treatments in chest wall irradiation after mastectomy and determine the potential benefit of flattening filter free beams. METHODS AND MATERIAL: Planning CT scans of 10 women requiring post-mastectomy chest wall radiotherapy were included in the study. A dose of 50 Gy in 2 Gy fractions was prescribed. Organs at risk (OARs) delineated were heart, lungs, contralateral breast, and spinal cord. Dose-volume metrics were defined to quantify the quality of concurrent treatment plans assessing target coverage and sparing of OARs. Plans were designed for conformal 3D therapy (3DCRT) or for RapidArc with double partial arcs (RA). RapidArc plans were optimized for both conventional beams as well as for unflattened beams (RAF). The goal for this planning effort was to cover 100% of the planning target volume (PTV) with ≥ 90% of the prescribed dose and to minimize the volume inside the PTV receiving > 105% of the dose. The mean ipsilateral lung dose was required to be lower than 15 Gy and V(20 Gy) < 22%. Contralateral organ irradiation was required to be kept as low as possible. RESULTS: All techniques met planning objectives for PTV and for lung (3DCRT marginally failed for V(20 Gy)). RA plans showed superiority compared to 3DCRT in the medium to high dose region for the ipsilateral lung. Heart irradiation was minimized by RAF plans with ~4.5 Gy and ~15 Gy reduction in maximum dose compared to RA and 3DCRT, respectively. RAF resulted in superior plans compared to RA with respect to contralateral breast and lung with a reduction of ~1.7 Gy and 1.0 Gy in the respective mean doses. CONCLUSION: RapidArc treatment resulted in acceptable plan quality with superior ipsilateral tissue sparing compared to traditional techniques. Flattening filter free beams, recently made available for clinical use, might provide further healthy tissue sparing, particularly in contralateral organs, suggesting their applicability for large and complex targets.

Studies on the defluoridation of water using conducting polymer/montmorillonite composites.

Conducting polymer/inorganic hybrid composites have large surface areas, which makes the adsorbent properties of the polymer composites as good the constituents. Polyaniline/montmorilonite (PANi-MMT) and polypyrrole/montmorillonite (PPy-MMT) composites were prepared, characterized (Fourier transform infrared, scanning electron microscopy, energy dispersive spectroscopy and X-ray diffraction patterns) and were employed as adsorbents for the removal of fluoride ions from aqueous solution by the batch sorption method. The spectral studies of the adsorbents before and after the adsorption are recorded to get better insight into the mechanism of the adsorption process. The results indicated that the removal of fluoride ions from water by these composites occurs via the combined effect of both the constituents, that is, through a physico-chemical mechanism. The amount of fluoride ion adsorbed by PANi-MMT and PPy-MMT at 30 degrees C is observed to be 2.3 and 5.1 mg g(-1), respectively, when compared to 0.77 and 2.66 mg g(-1), respectively, for the polymers alone. The Langmuir, Freundlich and Dubinin-Radushkevich isotherms were used to describe the adsorption equilibrium.

Loss of Function of OsFBX267 and OsGA20ox2 in Rice Promotes Early Maturing and Semi-Dwarfism in γ-Irradiated IWP and Genome-Edited Pusa Basmati-1.

Targeted mutagenesis is now becoming the most favored methodology to improve traits in popular rice cultivars selectively. Understanding the genetic basis of already available mutants could be the first step in designing such experiment. Improved White Ponni (IWP), a popularly grown South Indian rice variety, was subjected to γ irradiation to develop WP-22-2, an M6 line superior in semi-dwarfism, early flowering, and high yield, and it has grain qualities similar to those of IWP. The exogenous application of gibberellic acid (GA3) on WP-22-2 resulted in the elongation of shorter internodes to a level similar to IWP. The expression profiling of six genes regulating plant height showed their differential expression pattern at different time points post GA3 treatment. Furthermore, the sequencing of WP-22-2 and IWP genomes revealed several single nucleotide polymorphisms (SNPs) and large-scale deletions in WP-22-2. The conversion of functional codons to stop codons was observed in OsGA20ox2 and OsFBX267, which have been reported to have roles in regulating semi-dwarfism and early flowering, respectively. The loss of function of OsGA20ox2 and OsFBX267 in WP-22-2 resulted in reduced plant height as well as early flowering, and the same has been confirmed by editing OsGA20ox2 in the rice variety Pusa Basmati1 (PB1) using the CRISPR-Cas9 approach. The targeted editing of OsGA20ox2 in PB1 conferred shorter plant height to the edited lines compared with the wild type. Altogether, the study provides evidence on mutating OsGA20ox2 and OsFBX267 genes to develop early maturing and semi-dwarf varieties that can be released to farmers after functional characterization and field trials.

Comparative Analysis of Chitin SynthaseA dsRNA Mediated RNA Interference for Management of Crop Pests of Different Families of Lepidoptera.

RNA interference (RNAi) is a sequence-specific down-regulation in the expression of a particular gene, induced by double-stranded RNA (dsRNA). Feeding of dsRNA either directly or through transgenic plants expressing dsRNA of insect genes has been proven successful against lepidopteran and coleopteran pests, establishing an additional alternative to control insect pests. Lepidopteran crop pests including Spodoptera litura (Fabricius) (Noctuidae), Chilo partellus (Swinhoe) (Crambidae), Plutella xylostella (Linnaeus) (Plutellidae), and Maruca vitrata (Fabricius) (Pyralidae) are the devastating pests of a variety of crops. To tap the potential of RNAi against insect pests, a gene coding for the key enzyme in chitin biosynthesis in arthropods, the chitin synthaseA (CHSA), has been targeted through an exogenous delivery of dsRNA and plant-mediated RNAi. The introduction of dsCHSA caused "Half ecdysis" and "Black body" type lethal phenotypes and a significant reduction in larval body weight. Subsequent RT-qPCR analysis demonstrated the down-regulation of CHSA gene transcripts from 1.38- to 8.33-fold in the four target species. Meanwhile, when S. litura larvae fed with leaves of transgenic tobacco plants expressing dsSlCHSA, the mRNA abundance of CHSA gene was significantly decreased resulting in lethal phenotypes like "Double head formation," "Half ecdysis," and "Black body." In addition, abnormalities in pupal-adult and adult stage were also documented, strongly suggesting the RNAi effect of CHSA gene at late developmental stages. Overall, the results demonstrated that CHSA gene expression in Lepidopteran crop pests could be suppressed by application of dsRNA either as feeding or through transgenic crop plants.

Transgenic resistance by N gene of a Peanut bud necrosis virus isolate of characteristic phylogeny.

The nucleocapsid protein (N) gene of a Tospovirus devastating tomato crop in the south Indian state of Tamil Nadu was cloned and characterized. The high identity of the cloned sequence to a Peanut bud necrosis virus (PBNV) tomato isolate (97.8/99.6% nucleotide/amino acid) and a PBNV peanut isolate (94.4/96.3% nucleotide/amino acid) identified the Tospovirus as an isolate of PBNV, designated PBNV Coimbatore tomato (PBNV CT) isolate. Phylogenetic analysis of PBNV CT N gene provided useful insights into the movement and evolution of PBNV within Indian Territory. The characteristic phylogeny of PBNV CT N gene implied its potential to be an efficient transgene to confer effective PBNV resistance on crop plants. The efficacy of PBNV CT N gene in conferring PBNV resistance was studied by generating tobacco (Nicotiana tabacum L. cv Wisconsin) lines transgenic to the sense or antisense version of the gene. Several transgenic lines showed transgenic mRNA and/or protein accumulation, ranging from very high to undetectable levels, accompanied by different degrees of PBNV resistance. The undetectable or very low levels of transgene transcripts in certain PBNV-resistant sense or antisense N gene transgenic lines suggested RNA-mediated resistance by post-transcriptional gene silencing (PTGS) mechanism. However, PBNV resistance of certain transgenic lines with high levels of N gene transcripts was suggestive of possible operation of RNA-mediated non-PTGS mechanism(s) of resistance in those lines. Moreover, the high levels of N protein in certain PBNV-resistant sense N gene transgenic lines suggested protein-mediated resistance. The results predict the potential of PBNV CT N gene to confer effective PBNV resistance on tomato and other economically important crops.

A functional Ag-TiO2 nanocomposite solar selective absorber with antimicrobial activity by photochemical reduction process.

A functional nanocomposite coating developed by wet chemical route in a photochemical reduction process will be a good candidate for low temperature solar thermal application. The low-temperature curable cermet comprises of two different type of nanoparticles, generated by photocatalytic reduction of silver ions under the sunlight with the aid of crystalline TiO2 (~8 nm) nanoparticles. The optimized base absorber layer of Ag-TiO2 nanocomposite exhibited absorptance (α) of 0.90 and emittance (ε) of 0.18 at 200 °C. Further to improve the optical properties a second layer of MgF2 has been added to get high selective (α/ε = 0.93/0.19 at 200 °C) nature. In addition, the coating was demonstrated to possess an antimicrobial activity against both Gram-negative and Gram-positive bacteria. The presented work will provide a new insight into the spectrally selective absorbers and its antimicrobial nature, which may useful for water disinfection, hot water, industrial heating and swimming pool applications.

Genetic transformation of tropical maize (Zea mays L.) inbred line with a phytase gene from Aspergillus niger.

A full-length cDNA of phyA gene of Aspergillus niger, encoding phytase enzyme, was cloned and expressed in E. coli BL21 cells and assayed for its activity. The phyA cDNA consisted of 1404 bp, which encoded 467 amino acid residues. The phytase activity of purified phytase was 826.33 U/mL. The phyA gene under the control of endosperm-specific promoters was transformed into an Indian maize inbred line, UMI29, using particle bombardment-mediated transformation method to generate transgenic maize plants over-expressing phytase in seeds. PCR and GUS analyses demonstrated the presence of transgenes in T0 transgenic plants and their stable inheritance in the T1 progenies. Three transgenic events expressing detectable level of A. niger phytase were characterized by western blot analysis. Phytase activity of 463.158 U/kg of seed was observed in one of the events, JB-UMI29-Z17/2. The phytase activity of transgenic maize seeds was 5.5- to 7-fold higher than the wild-type UMI29 seeds and, consequently, the seeds had 0.6- to 5-fold higher inorganic phosphorus content.

Sardine oil loaded vanillic acid grafted chitosan microparticles, a new functional food ingredient: attenuates myocardial oxidative stress and apoptosis in cardiomyoblast cell lines (H9c2).

Fish oil has been widely recognized as an excellent dietary source of polyunsaturated n-3 fatty acids such as EPA and DHA. However, it can undergo oxidation easily resulting in the formation of toxic off flavor compounds such as hydroperoxides. These compounds adversely affect the nutritional quality and may induce several stress reactions in body. To solve this problem, a new antioxidant bio-material, vanillic acid-grafted chitosan (Va-g-Ch), was synthesized and used as a wall material for microencapsulation of fish oil. The sardine oil loaded Va-g-Ch microparticles could be a potential functional food ingredient considering the numerous health benefits of fish oil, chitosan, and vanillic acid. The current study aimed to investigate the possible protective effect of sardine oil-loaded Va-g-Ch microparticles against doxorubicin-induced cardiotoxicity and the underlying mechanisms. In vitro cytotoxicity evaluation was conducted using H9c2 cardiomyocytes. MTT assay revealed that effective cytoprotective effect was induced by a sample concentration of 12.5 µg/mL. Results of apoptosis by double fluorescent staining with acridine orange/ethidium bromide and caspase-3 evaluation by ELISA substantiated the above findings. Further, flow cytometric determination of membrane potential, relative expression of NF-κB by PCR, and ROS determination using DCFH-DA also confirmed the protective effect of encapsulated sardine oil against doxorubicin-induced cardiotoxicity. NF-κB expression was down-regulated nearly by 50% on cells treated with encapsulated sardine oil. Altogether, the results revealed that sardine oil-loaded Va-g-Ch microparticles demonstrated potential cell protection against doxorubicin-induced oxidative stress.

Beaded reactive polymers. 3. Effect of triacrylates as crosslinkers on the physical properties of glycidyl methacrylate copolymers and immobilization of penicillin G acylase.

Various glycidyl methacrylate (GMA) copolymers were synthesized by suspension polymerization, using pentaerythritol triacrylate (PETA), trimethylolpropane triacrylate (TMPTA), and trimethylolpropane trimethacrylate (TRIM) as crosslinking comonomers. These copolymers were evaluated for the immobilization of penicillin G acylase. Broad pore-size distribution that was observed was in the range 5-300 nm. Both surface area and pore volume increased with increase in the mole fraction of crosslinking comonomer (increasing crosslink density). The pore volume of the copolymers was more than doubled by including lauryl alcohol as porogen. Binding of penicillin G acylase (PGA) was quantitative on highly crosslinked copolymers. The expression of bound PGA was better on the relatively more hydrophilic GMA-TMPTA and GMA-PETA copolymer supports compared to the GMA-TRIM copolymers. Among the different copolymers studied, GMA-TMPTA copolymer 7411 exhibited highest activity of immobilized penicillin G acylase (167.4 IU/g) with 35.1% expression.

Adsorption and expression of penicillin G acylase immobilized onto methacrylate polymers generated with varying pore generating solvent volume.

Adsorption and expression of penicillin G acylase was studied on macroporous methacrylate polymer beads of differing pore volume, generated with kerosene. The absorption and expression of the penicillin G acylase was dependent on pore volume. Maximum expression of 57% of adsorbed enzyme was obtained on beads synthesized with 40 mL of kerosene, indicating minimum pore-diffusion limitations.

Immobilization of penicillin G acylase on methacrylate polymers.

Macroporous weak cation-exchange methacrylate polymers were synthesized for the immobilization of penicillin G (Pen G) acylase. The role of certain factors such as pore-generating solvent, cross-linking agent, cross-linking density, and comonomer, in enzyme adsorption and expression was studied. Kerosene was a superior pore-generating solvent to paraffin oil. Ethylene glycol dimethacrylate and acrylic acid served as the best cross-linking agent and comonomer, respectively, in the systems studied. 80.3% of the activity of the enzyme adsorbed onto polymer beads prepared with 0.05 mol of acrylic acid (polymer PM-39) was expressed. Properties of the Pen G acylase, immobilized on PM-39 by adsorption and cross-linking with glutaraldehyde (IME-PM-39) were studied. The optimum pH, optimum temperature and Km of Pen G acylase shifted from 8.0 to 7.5-7.8, 50 degrees C to 55 degrees C and 0.038 mol dm-3 to 2.4-3.0 mol dm-3, respectively, as a result of immobilization on PM-39. IME-PM-39 was used repeatedly for 15 cycles in the production of 6-amino penicillanic acid (6-APA).

Bactericidal power of neutrophils in protein calorie malnutrition.

The present study was undertaken to compare the disturbance in bactericidal power of neutrophils in 25 children having protein caloric malnutrition age and sex matched healthy control subjects. There was a highly significant reduction in the percentage of bacteria killed during incubation in children having PCM as compared to healthy control. A direct relationship existed between total serum proteins and bactericidal activity of neutrophils in children having PCM. The impaired bactericidal power of neutrophils can be attributed to impaired synthesis of lysosomal enzymes, glycolytic activity of neutrophils in children having PCM. The exact mechanisms still remain to be elucidated. Decreased bactericidal activity of neutrophils may be one of the mechanisms responsible for infection.

A study of incidence of Australia antigen and derangements in liver function tests in leprosy.

The present study was conducted in 50 patients of various subtypes of leprosy (Lepromatous, Tuberculoid, Borderline borderline) and 25 healthy control, for detection of Australia antigen and various liver function tests (serum protein, cholesterol, alkaline phosphates, SGOT, SGPT, bilirubin and liver biopsy) to see incidence of Australia Antigen and derangement in liver function. It was concluded that incidence of Australia antigen in study and control group was zero. Total serum protein and serum globulin was increased in lepromatous leprosy. A/G ratio was reversed in 34.3% and 50% in lepromatous and tuberculoid leprosy respectively. Granulomatous hepatitis was seen in 66.66% and 50% cases of lepromatous and tuberculoid leprosy respectively. No relationships was established between hepatic lesion, Australia antigen and liver function test.

Klippel-Trenaunay syndrome and gestational trophoblastic neoplasm.

BACKGROUND: Klippel-Trenaunay syndrome is a non-heritable venous malformation with bone and soft tissue hypertrophy and cutaneous nevi. CASE CHARACTERISTICS: Neonate with Klippel Trenaunay syndrome born to a mother with past history of Gestational trophoblastic neoplasm. OBSERVATION: Antenatally, a fetal vascular malformation was identified ultrasonologically at 29 weeks gestation. Acute myeloid leukemia was diagnosed in mother at 33 weeks gestation. MESSAGE: A rare association of Klippel Trenaunay syndrome and gestational trophoblastic neoplasm with the possible role of either hyperglycosylated Human Chorionic Gonadotropin or chemotherapy as a link is highlighted.