Single-drop and nanogram level determination of sulfite (SO3(2-)) in alcoholic and nonalcoholic beverage samples based on diffuse reflectance fourier transform infrared spectroscopic (DRS-FTIR) analysis on KBr matrix

The diffuse reflectance Fourier transform spectroscopic (DRS-FTIR) method, using potassium bromide matrix, has been developed for the one-drop microdetermination of sulfite in beverage samples. The present method is very simple, rapid, and precise for the determination of sulfite. The nanogram level of sulfite determination is based on the selection of a quantitative analytical peak at 495 cm (-1) among the three observed vibrational peaks obtained by diffuse reflectance Fourier transform infrared spectroscopy (DRS-FTIR). As little as a single drop of sample is sufficient for quantitative analysis of sulfite. The limit of detection (LOD) and the limit of quantification (LOQ) of the method are found to be 8 and 40 ng of SO 3 (2-) 0.1 g (-1) of KBr matrix, respectively. The linear range of the method (LR) as well as the LOD based on the concentration of sulfite in the solution are 5-500 and 0.8 microg/mL, respectively. The precision in terms of standard deviation and relative standard deviation value at a level of 100 ng of SO 3 (2-) 0.1 g (-1) of KBr for n = 10 are found to be 2 ng of SO 3 (2-) and 2.3%, respectively. The relative standard deviation ( n = 10) for the determination of sulfite in beverage samples available in the local market was observed to be in the range of 2.4-7.8%. The method is free from interionic effects of foreign species. No sample pretreatment is required in this method. The proposed method avoids the requirement of large numbers and bulk amounts of reagents. The method is well-suited to the need of green chemistry.     

Conservation assessment and prioritization of areas in Northeast India: Priorities for amphibians and reptiles

This study combines niche modeling and systematic area prioritization using distribution data for 131 species of amphibians and reptiles from Northeast India and Burma, with two objectives: (i) to evaluate the performance of the current conservation area network in Northeast India with respect to the representation of amphibians and reptiles, and (ii) to identify potential areas for expanding the current conservation area network. In a two-step protocol, maximum entropy niche modeling was used to project species’ potential geographic occurrences, and the resulting probabilistic distribution data were used to prioritize areas with algorithms that maximize the representation of all species in minimal total area. The results provided a critical assessment of conservation priorities in this data-deficient region, and indicate the utility of combining niche modeling with systematic area prioritization in such situations. Many areas that had been overlooked in previous assessments were identified. Although the existing protected areas were found to be inadequate for representation of amphibian and reptile diversity, the prioritization results show that by targeting a minimal representation of 5% of the current total area suitable for each species, the gaps can be filled with a relatively modest (0.41%) increase in the current total area covered by the network. Extended analyses were also performed to assess the effects of putatively rare species on reserve selection, which showed that the inclusion of these taxa can change the prioritization solutions significantly. The prioritization results also highlight areas of Northeast India that warrant attention from future surveys.     

Eastern Indian 3800-million-year-old crust and early mantle differentiation

Samarium-neodymium data for nine granitic and tonalite gneisses occurring as remnants within the Singhbhum granite batholith in eastern India define an isochron of age 3775 +/- 89 x 10(6) years with an initial (143)Nd/(144)Nd ratio of 0.50798 +/- 0.00007. This age contrasts with the rubidium-strontium age of 3200 x 10(6) years for the same suite of rocks. On the basis of the new samarium-neodynium data, field data, and petrologic data, a scheme of evolution is proposed for the Archean crust in eastern India. The isotopic data provide evidence that parts of the earth's mantle were already differentiated with respect to the chondritic samarium-neodymium ratio 3800 x l0(6) years ago.     

A functional screen for the type III (Hrp) secretome of the plant pathogen Pseudomonas syringae

Type III secreted "effector" proteins of bacterial pathogens play central roles in virulence, yet are notoriously difficult to identify. We used an in vivo genetic screen to identify 13 effectors secreted by the type III apparatus (called Hrp, for "hypersensitive response and pathogenicity") of the plant pathogen Pseudomonas syringae. Although sharing little overall homology, the amino-terminal regions of these effectors had strikingly similar amino acid compositions. This feature facilitated the bioinformatic prediction of 38 P. syringae effectors, including 15 previously unknown proteins. The secretion of two of these putative effectors was shown to be type III--dependent. Effectors showed high interstrain variation, supporting a role for some effectors in adaptation to different hosts.     

Les hydrolases du sol et leur association avec les hydrates de carbone

Soil hydrolases and their association with carbohydrates. Active enzymes isolated from soil in the form of carbohydrate-enzyme-complexes could not be separated from each other by DEAE cellulose chromatography. All enzymes measured in this complex were recovered in Chromatographie fractions with different specific activities and variations in carbohydrate contents. There is however, no linear relationship between the quantity of carbohydrates and specific activities of the fractions. The carbohydrate-enzyme-complexes are not glycoproteins, but the enzymes are physically adsorbed on polysaccharides in a non-covalent manner. The essential role of this carbohydrate is to protect the enzyme systems against denaturation or proteolysis, thus ensuring their stability in soil.     

Effect of inorganic nitrogen nutrition on cytokinin-induced potato microtuber production in vitro

Summary The effect of inorganic nitrogen nutrition on the induction and development of microtubers by cytokinin-induced tuberization was studied in four potato genotypes belonging to different maturity groups. The objective of this study was to investigate whether a reduction in total nitrogen level in the Murashige & Skoog medium would improve cytokinin-induced tuberization rate. The effect of three levels of total nitrogen (15, 30 and 45 meq) on tuberization was studied at constant (20 meq K) and varying potassium levels approximating to 5, 10 and 15 meq. Reducing the total nitrogen supply increased the number but decreased the size of nitrogen level on the rate of assimilate partitioning (harvest index) during cytokinin-induced microtuberization.     

HIV-1 proviral DNA excision using an evolved recombinase

HIV-1 integrates into the host chromosome and persists as a provirus flanked by long terminal repeats (LTRs). To date, treatment regimens primarily target the virus enzymes or virus-cell fusion, but not the integrated provirus. We report here the substrate-linked protein evolution of a tailored recombinase that recognizes an asymmetric sequence within an HIV-1 LTR. This evolved recombinase efficiently excised integrated HIV proviral DNA from the genome of infected cells. Although a long way from use in the clinic, we speculate that this type of technology might be adapted in future antiretroviral therapies, among other possible uses.     

Cytotoxicity of Senecio in macrophages is mediated via its induction of oxidative stress

In Arunachal Pradesh and other sub-Himalayan areas of India, accidental consumption of Senecio plants by yaks is often fatal as the plant contains toxic alkaloids like Seneciophylline. The present investigation was undertaken to demonstrate the pro-oxidant effects of an ethanolic extract of Seneciochrysanthemoides (S-EtOH). S-EtOH impaired viability in macrophages, the IC₅₀ being 13.8 ± 1.11 μg/mL. The effect of S-EtOH (1 μg/mL) on generation of reactive oxygen species (ROS) in macrophages was measured by flow cytometry using 2′,7′-dichlorofluorescein diacetate (H₂DCFDA) where it caused a significant increase in the mean fluorescence channel (MFC) from 8.55 ± 0.03 to 47.32 ± 2.25 (p < 0.001). S-EtOH also effected a 3.8-fold increase in extracellular nitric oxide (NO) generation from 4.90 ± 0.72 μM to 18.79 ± 0.32 μM (p < 0.001), a 2.2-fold increase in intracellular NO production, the MFC increasing from 14.95 ± 0.48 to 33.34 ± 1.66 (p < 0.001), and concomitantly depleted non protein thiols as analyzed by flow cytometry using mercury orange, with a reduction in MFC from 632.5 ± 49.44 to 407.4 ± 12.61 (p < 0.01). Additionally, S-EtOH (14 μg/mL, 24 h) caused apoptosis as evident by increased Annexin V binding and terminal deoxynucleotidyl transferase mediated dUTP DNA nick end labeling. Taken together, the cytotoxicity of S-EtOH can be partly attributed to its capacity to inflict oxidative damage via generation of both reactive oxygen and nitrogen species culminating in apoptosis.     

High-throughput approach to detection of knockdown resistance (kdr) mutation in mosquitoes, Culex quinquefasciatus, based on real-time PCR using single-labelled hybridisation probe/melting curve analysis

BACKGROUND: Knockdown resistance (kdr) mutation (L1014F) is a well‐defined mechanism of resistance to pyrethroids and DDT in many insect species. Sensitive detection of the mutations associated with resistance is a prerequisite for resistance management strategies. The authors have developed a new real‐time molecular diagnostic assay based on SimpleProbe^®/melting curve analysis for large‐scale kdr genotyping in the wild population of Culex quinquefasciatus Say, the principal vector of bancroftian filariasis. Melting curve analysis is based on the thermal stability difference between matched and mismatched DNA duplexes. The application of SimpleProbe^® chemistry in insects described here is novel in entomology research. RESULTS: The mosquitoes homozygous for knockdown‐resistant and knockdown‐susceptible allele showed melting peaks at 60.45 °C ( ± 0.25) and 64.09 °C ( ± 0.24) respectively. The heterozygous mosquitoes yielded both peaks at approximately 60.5 °C ( ± 0.2) and 64.20 °C ( ± 0.23). Among the 92 samples genotyped, 16 were found to be homozygous resistant, 44 homozygous susceptible and 32 heterozygous. Comparative assessments were made of all the reported methods for kdr genotyping. CONCLUSION: The present method is cheaper, faster, more reliable and versatile than other alternatives proposed in detecting correct kdr genotypes in mosquitoes. This is the first report using a single‐labelled hybridisation probe to detect point mutations in insect populations. Copyright © 2010 Society of Chemical Industry     

Evaluation of mulberry germplasm for resistance to powdery mildew in the field and greenhouse

Powdery mildew caused by the ascomycete Phyllactinia guttata (syn. P. corylea) is a major foliar disease worldwide of the unique mulberry (Morus spp.) for silkworm feed. Genetic resistance to powdery mildew, the most sustainable and economical strategy for disease control, is still elusive for tropical mulberry. About 147 germplasm sources, representing 18 countries of origin, were screened for resistance to P. guttata in six seasonal fields and greenhouse trials after exposure to natural and artificial inoculum, respectively. In the field, the level of plant responsiveness to disease was assessed from 30 to 62 days after pruning in each season as variations in the disease severity index (DSI), disease incidence (DI%) and area under the disease progress curve (AUDPC). These measures differed significantly among the germplasm. Of 147 germplasm sources, ~6.8% had useful resistance (two high and nine moderately resistant) to the powdery mildew pathogen on the basis of DSI. The AUDPC values were 13.5-fold higher in the most susceptible accession—(Philippines) than the least responsive (Vietnam-2). The results of DSI were strongly correlated with the obtained DI values (r = 0.92; P < 0.01) and AUDPC (r = 0.89; P < 0.01). Moreover, field screening results were highly correlated (R ² = 0.839) with values from the greenhouse evaluation using artificial inoculum. However, the DSI values in field and greenhouse screenings for three sources (Non-nayapati, Nao-khurkul and Tista Valley) varied significantly. A relatively low disease reaction of 09 resources (Vietnam-2, Ankara and 07 others) using different assessment scales after natural and artificial inoculation prove, for the first time, that they have potential in breeding for resistance in tropical mulberry to powdery mildew.