Ellagic acid mitigates arsenic‐trioxide‐induced mitochondrial dysfunction and cytotoxicity in SH‐SY5Y cells

In the current study, neuroprotective significance of ellagic acid (EA, a polyohenol) was explored by primarily studying its antioxidant and antiapoptotic potential against arsenic trioxide (As₂O₃)‐induced toxicity in SH‐SY5Y human neuroblastoma cell lines. The mitigatory effects of EA with particular reference to cell viability and cytotoxicity, the generation of reactive oxygen species, DNA damage, and mitochondrial dynamics were studied. Pretreatment of SH‐SY5Y cells with EA (10 and 20 μM) for 60 min followed by exposure to 2 μM As₂O₃ protected the SH‐SY5Y cells against the harmful effects of the second. Also, EA pre‐treated groups expressed improved viability, repaired DNA, reduced free radical generation, and maintained altered mitochondrial membrane potential than those exposed to As₂O₃ alone. EA supplementation also inhibited As₂O₃‐induced cytochrome c expression that is an important hallmark for determining mitochondrial dynamics. Thus, the current investigations are more convinced for EA as a promising candidate in modulating As₂O₃‐induced mitochondria‐mediated neuronal toxicity under in vitro system.     

Three dimensional (3D) structure prediction and substrate-protein interaction study of the chitin binding protein CBP24 from B. thuringiensis

Bacillus thuringiensis is an insecticidal bacterium whose chitinolytic system has been exploited to improve insect resistance in crops. In the present study, we studied the CBP24 from B. thuringiensis using homology modeling and molecular docking. The primary and secondary structure analyses showed CBP24 is a positively charged protein and contains single domain that belongs to family CBM33. The 3D model after refinement was used to explore the chitin binding characteristics of CBP24 using AUTODOCK. The docking analyses have shown that the surface exposed hydrophilic amino acid residues Thr-103, Lys-112 and Ser-162 interact with substrate through H-bonding. While, the amino acids resides Glu-39, Tyr-46, Ser-104 and Asn-109 were shown to have polar interactions with the substrate. The binding energy values evaluation of docking depicts a stable intermolecular conformation of the docked complex. The functional characterization of the CBP24 will elucidate the substrate-interaction pathway of the protein in specific and the carbohydrate binding proteins in general leading towards the exploration and exploitation of the prokaryotic substrate utilization pathways.     

Ethnobotanical assessment of plant resources of Banda Daud Shah,District Karak,Pakistan

Background

The Indigenous knowledge of plants is scientifically and culturally very significant. This paper elucidates the empirical findings of an ethnobotanical survey of Banda Daud Shah, District Karak, Pakistan.

Methods

Data collection was carried out from October 2011 to September 2012. Total twelve survey trips were made, three in each season. About 100 respondents were interviewed; most of them were aged people between 60–70 years. Interviews were conducted using structured questionnaire composed of variety of questions regarding ethnomedicinal uses of plants of the study area. Direct matrix ranking (DMR), informant citations and market survey of multipurpose plants were also carried out.

Results

The local community was using 58 plant species belonging to 52 genera and 34 families for different purposes. A total of 25 plant species were herbs followed by 18 shrubs. Leaf (45%) was the most commonly used plant part followed by the whole plants (23%). In total, 40 plant species were medicinally used to treat variety of diseases, of which highest number of species being used for gastro-intestinal problems (19 spp.), expectorant (3 spp.) and antipyretic (3 spp.). Beside medicinal values, 25 species were used for fuel and 18 for fodder purposes. Informant consensus showed that gastrointestinal and respiratory infections were ranked highest (F_IC?=?0.75) among all ailments. According to DMR output, Dalbergia sisso ranked first due to high multipurpose uses among all species and was found most threatened with higher market value.

Conclusion

The investigated area is rural in nature and the inhabitants are highly dependent on the native plants for their health care needs and other requirements like fuel wood and fodder due to financial constraints and unavailability of resources. Medicinal plants for high ranked diseases may be phtyochemicaly and pharmacologically investigated to prove their efficacy. The local medicinal flora is facing overexploitation, overgrazing and improper way of collection. Proper conservation strategies such as controlled grazing, reforestation and rangeland management among many others may be adopted to promote the sustainable use of medicinal plants.

     

Carandinol: First isohopane triterpene from the leaves of Carissa carandas L. and its cytotoxicity against cancer cell lines

A new triterpene carandinol (1) was isolated from the leaves of Carissa carandas L., along with five known compounds, betulinic acid (2), β-sitosterol-3-O-β-d-glucopyranoside (3), oleanolic acid (4), ursolic acid (5) and 4-hydroxybenzoic acid (6). The structure of compound 1 was deduced as 3β,21α-dihydroxyisohopane by exhaustive spectroscopic analyses. The known compounds 2–6 were identified by comparison with the reported spectral data. Compound 1 was evaluated for cytotoxicity, immunomodulatory, antiglycation, antioxidant and enzyme inhibition activity. It exhibited significant in vitro cytotoxicity to every cell line tested (HeLa, PC-3 and 3T3) and was relatively more toxic to human cervical cancer (HeLa) cell line. This is the first report of the isolation of an isohopane triterpene from the genus Carissa. Carandinol also represents the first example of a cytotoxic isohopane.     

Applications of Decision Support System: A Case Study of Solanaceous Vegetables

Crop simulation models constitute the major proportion in decision support systems. A large number of crop models have been developed for potato and few for tomato and peppers. In the literature, thirty three crop models have been reported to simulate potato, nine for tomato and six for peppers. Some of these models dealt with the climate change scenario and others with the crop management practices such as sowing time, irrigation, nitrogen, and insect-pests management. The most evaluated and applied models for potato include; SUBSTOR, and LINTUL-Potato, whereas CROPGRO-tomato model is the most tested and applied for tomato. The AQUACROP is the most widely used model to simulate the water dynamics. The CROPGRO model has been tested for elevated temperatures and CO₂ under greenhouse conditions for tomato. In tomato and peppers, almost similar models have been applied for field conditions as well as under greenhouse environments with some modifications. Nitrogen dynamics has been widely tested by employing the EU-Rotate-N model for tomato and peppers. Simulation studies dealing with changing climate conditions are rare in potato and are not found for tomato and peppers. To modify potato, tomato and peppers models for climate impact studies, it is required that they are (a) calibrated and evaluated with new cultivars under various agro-environmental conditions and (b) assessed under varying field conditions under changing climates and crop management practices, including temperature increases, water and nutrient management and their interactions. These comprehensive model studies and modifications need a collaborative international effort and a multi-year, large scale field research studies on potato, tomato and peppers.     

Inferring the contribution of sexual reproduction,migration and off‐season survival to the temporal maintenance of microbial populations: a case study on the wheat fungal pathogen Puccinia striiformis f.sp. tritici

Understanding the mode of temporal maintenance of plant pathogens is an important domain of microbial ecology research. Due to the inconspicuous nature of microbes, their temporal maintenance cannot be studied directly through tracking individuals and their progeny. Here, we suggest a series of population genetic analyses on molecular marker variation in temporally spaced samples to infer about the relative contribution of sexual reproduction, off‐season survival and migration to the temporal maintenance of pathogen populations. We used the proposed approach to investigate the temporal maintenance of wheat yellow rust pathogen, Puccinia striiformis f.sp. tritici (PST), in the Himalayan region of Pakistan. Multilocus microsatellite genotyping of PST isolates revealed high genotypic diversity and recombinant population structure across all locations, confirming the existence of sexual reproduction in this region. The genotypes were assigned to four genetic groups, revealing a clear differentiation between zones with and without Berberis spp., the alternate host of PST, with an additional subdivision within the Berberis zone. The lack of any differentiation between samples across two sampling years, and the very infrequent resampling of multilocus genotypes over years at a given location was consistent with limited over‐year clonal survival, and a limited genetic drift. The off‐season oversummering population in the Berberis zone, likely to be maintained locally, served as a source of migrants contributing to the temporal maintenance in the non‐Berberis zone. Our study hence demonstrated the contribution of both sexual recombination and off‐season oversummering survival to the temporal maintenance of the pathogen. These new insights into the population biology of PST highlight the general usefulness of the analytical approach proposed.     

Polyphosphate Deficiency in Mycobacterium tuberculosis Is Associated with Enhanced Drug Susceptibility and Impaired Growth in Guinea Pigs

Inorganic polyphosphate (polyP), a linear polymer of hundreds of phosphate residues linked by ATP-like phosphoanhydride bonds, is found in all organisms and performs a wide variety of functions. This study shows that polyP accumulation occurs in Mycobacterium tuberculosis upon exposure to various stress conditions. M. tuberculosis possesses a single homolog of ppk-1, and we have disrupted ppk-1 in the M. tuberculosis genome by allelic replacement. The mutant strain exhibited negligible levels of intracellular polyP, decreased expression of sigF and phoP, and reduced growth in the stationary phase and displayed a survival defect in response to nitrosative stress and in THP-1 macrophages compared to the wild-type strain. We report that reduction in polyP levels is associated with increased susceptibility of M. tuberculosis to certain TB drugs and impairs its ability to cause disease in guinea pigs. These results suggest that polyP contributes to persistence of M. tuberculosis in vitro and plays an important role in the physiology of bacteria residing within guinea pigs.     

Silicon-mediated growth and yield improvement of sunflower (<Emphasis Type="Italic">Helianthus annus</Emphasis> L.) subjected to brackish water stress

Silicon is known to compensate crop yield losses under diverse biotic and abiotic stress conditions; however, reports about its protective role for plants exposed to brackish water stress are very limited. A pot culture experiment was conducted to assess the beneficial effect of silicon supplementation (0 and 100 mg/kg) in alleviating growth adversities of brackish water (saline, sodic, alkaline, and saline–sodic water) stress in two contrasting sunflower cultivars, SF-187 (salt tolerant), and Hysun-33 (salt sensitive) grown in greenhouse. Results demonstrated that hostile growth environments, mainly the combined stress of saline–sodic water, severely affected the physiological attributes, growth, yield, and yield contributing components in sunflower. However, the response to brackish water stress differed genotypically, with greater magnitude of damage to the Hysun-33 as compared to SF-187 genotype. It hampered plant growth due to membrane damage and reduced water uptake, but silicon supplementation minimized the negative effects of stress by limiting toxic Na⁺ ions uptake, improving membrane stability, and increasing relative water contents caused by higher silicon and K⁺ uptake that eventually led to improved biomass yield. The response was further evaluated at yield level and data regarding head diameter, achene yield, and 100 achene weight were taken. Results indicated that silicon supplementation to growth medium of saline and/or sodic water treated plants significantly enhanced the head diameter (22–30%), thus ultimately producing 15–25% higher achene yield, and weight of the biological harvest of both sunflower genotypes. Overall, the beneficial effect of silicon supplementation was more evident in Hysun-33 (salt sensitive) as compared to SF-187 (salt tolerant) genotype. Taken together, the results of this study suggest silicon fertilization as a potential strategy to increase crop productivity under brackish water stress; however, experimental trials at farmer field level should be conducted before setting any recommendations.     

Purification and characterization of a high-molecular-weight form of recombinant human Interleukin-2

During purification of recombinant Interleukin-2 (rIL-2) by reversed-phase HPLC, early fractions are discarded due to the presence of an unidentified form of rIL-2. A procedure has been developed to isolate and purify this unidentified form of rIL-2. The purification process involves two chromatography steps and utilizes a Bakerbond Carboxy-Sulfon (CS) column under two different conditions. This material, designated as a high-molecular-weight form of rIL-2 (HMWrIL-2), exhibits lower mobility during SDS-PAGE and has apI which is approximately one unit less than that of rIL-2, but has similar bioactivity to rIL-2. Structural analysis through enzymatic cleavage, HPLC peptide mapping, mass spectrometry, sequencing, and amino acid composition revealed that the difference between these two proteins is a C-terminal extension of 11 amino acids. This extension could be the result of a nonstandard translation event.