Differential HSP90α expression in fish hepatocytes from polluted estuary during summer

The molecular chaperone, heat shock protein 90α (HSP90α), plays an important role in protein folding, degradation of denatured proteins and steroid activation. It is essential for the maintenance of cellular integrity and survival when induced in response to environmental, physical and chemical stresses. In the present investigation the effect of environmental stress on HSP90α expression was examined in grey mullet Mugil cephalus living in either a contaminated (Ennore) or uncontaminated (Kovalam) estuary over two seasons: Hepatocytes were isolated from grey mullet of both estuaries. Oxidative stress was determined along with HSP90α in these fish. Additionally, immunohistochemical changes were studied to confirm the HSP90α expression. Comparison of the results revealed enhanced hepatocyte oxidative stress and HSP90α expressions in fish from Ennore to a significant extent than fish from Kovalam. Also, the results showed significant seasonal variations with maximum expression observed during summer compared to the monsoon season. Overexpression of HSP90α in hepatocytes exposed to chronic environmental stress by pollutants may confer differential effects on cell survival by protecting against oxidative stress induced changes. The results also indicate that seasonal variations have significant effect on the HSP90α expression.     

Marker assisted accelerated introgression of null allele of kunitz trypsin inhibitor in soybean

Development of kunitz trypsin inhibitor (KTI)-free soybean is crucial for soy-food industry as the heat inactivation employed to inactivate the anti-nutritional factor in regular soybean incurs extra cost and affects protein solubility. In the presented work, a null allele of KTI from PI542044 was introgressed into cultivar ‘JS97-52’ (recurrent parent) through marker assisted backcrossing. Foreground selection in BC₁F₂, BC₂F₂ and BC₃F₂ was carried out using the null allele-specific marker in tandem with SSR marker Satt228, tightly linked with a trypsin inhibitor Ti locus. Background selection in null allele-carrying plants through 106 polymorphic SSR markers across the genome led to the identification of 9 KTI-free lines exhibiting 98.6% average recurrent parent genome content (RPGC) after three backcrosses, which otherwise had required 5–6 backcrosses through conventional method. Introgressed lines (ILs) were free from KTI and yielded at par with recurrent parent. Reduction of 68.8–83.5% in trypsin inhibitor content (TIC) in ILs compared to the recurrent parent (‘JS97-52’) was attributed to the elimination of KTI.     

Genetic diversity in <Emphasis Type="Italic">Eremostachys superba</Emphasis> Royle ex Benth. (Lamiaceae), an endangered Himalayan species,as assessed by RAPD

Eremostachys superba Royle ex Benth. (Lamiaceae) has undergone a severe decline in population size since its discovery in the North-western Himalayas in late 19th century. One hundred and seventy-two plants from six populations in the Indian states of Uttar Pradesh and Jammu & Kashmir, located between 0.45 km and 455.72 km apart from each other were evaluated for RAPD polymorphism. Sixteen random primers generated 92 bands overall, 77 of which were polymorphic. Shannon’s index of genetic diversity within populations (H _o) ranged between 0.305 and 0.421; the average within-population diversity (H _pop) was 0.389; and the total species diversity (H _sp) was 0.478. The population from Mohand (representing the type locality) had the fewest plants, at 18, and was genetically the most depauperate. Among the other populations, ranging in size between 52 and 1,022 individuals, no relation between population size and genetic diversity was evident. It is suggested that these six populations represent relics of a larger, extended population, in which the presence of perennating rootstocks has helped preserve historic patterns of genetic diversity. AMOVA revealed that 83.01% of the variation exists within populations, which was consistent with earlier studies on the reproductive biology of E. superba, which indicated this species is predominantly allogamous. FST distances between all populations were significant, indicating geographic differentiation despite some of them being closely separated. Habitat restoration and protection from indiscriminate harvesting are proposed as primary strategies for conserving E. superba. Rejuvenation of the Mohand population through intrapopulation crossing between plants bearing diverse molecular phenotypes is also suggested.     

RAPD and ISSR fingerprinting in cultivated chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) and its wild progenitor <Emphasis Type="Italic">Cicer reticulatum</Emphasis> Ladizinsky

Detection of genetic relationships between 19 chickpea cultivars and five accessions of its wild progenitor Cicer reticulatum Ladizinsky were investigated by using RAPD and ISSR markers. On an average, six bands per primer were observed in RAPD analysis and 11 bands per primer in ISSR analysis. In RAPD, the wild accessions shared 77.8% polymorphic bands with chickpea cultivars, whereas they shared 79.6% polymorphic bands in ISSR analysis. In RAPD analysis 51.7% and 50.5% polymorphic bands were observed among wild accessions and chickpea cultivars, respectively. Similarly, 65.63% and 56.25% polymorphic bands were found in ISSR analysis. The dendrogram developed by pooling the data of RAPD and ISSR analysis revealed that the wild accessions and the ICCV lines showed similar pattern with the dendrogram of RAPD analysis. The ISSR analysis clearly indicated that even with six polymorphic primers, reliable estimation of genetic diversity could be obtained, while nearly 30 primers are required for RAPD. Moreover, RAPD can cause genotyping errors due to competition in the amplification of all RAPD fragments. The markers generated by ISSR and RAPD assays can provide practical information for the management of genetic resources. For the selection of good parental material in breeding programs the genetic data produced through ISSR can be used to correlate with the relationship measures based on pedigree data and morphological traits to minimize the individual inaccuracies in chickpea.     

The role of technology in enhancing low resource agriculture in Africa

Traditional forms of farming, herding, and fishing are remarkably adapted to African conditions but these traditional approaches are being overtaken by modern pressures, particularly population growth. According to a report published by the Office of Technology Assessment (OTA), a nonpartisan analytical support agency of the U. S. Congress, one promising way to help African farmers and herders would be for development assistance organizations to focus more attention on the various forms of low-resource agriculture that predominate in Africa. In keeping with OTA's mission and primary audience, “Enhancing Agriculture in Africa: A Role for U. S. Development Assistance” (1988) is a policy-oriented synthesis of available technical information. The report provides Congress with a range of options that, if pursued, could help Africans enhance agriculture, increase their food security, and improve their lives. This paper is drawn from the larger OTA report, and it focuses on the role technology might play in enhancing low-resource agriculture. Readers should see the full assessment (OTA, 1988) for more information on policy considerations; the specific technologies mentioned; or a complete list of advisory panel members, workshops and participants, and commissioned papers. OTA's report comes at a critical time: for a variety of reasons—ranging from changing values to increased budget constraints—U. S. foreign assistance policy is undergoing a fundamental reevaluation. This review of the potential of low-resource agriculture, and options the United States might pursue to enhance this approach, was intended to aid in this reevaluation.     

Amplification of acetylcholine-binding catenanes from dynamic combinatorial libraries

Directed chemical synthesis can produce a vast range of molecular structures, but the intended product must be known at the outset. In contrast, evolution in nature can lead to efficient receptors and catalysts whose structures defy prediction. To access such unpredictable structures, we prepared dynamic combinatorial libraries in which reversibly binding building blocks assemble around a receptor target. We selected for an acetylcholine receptor by adding the neurotransmitter to solutions of dipeptide hydrazones [proline-phenylalanine or proline-(cyclohexyl)alanine], which reversibly combine through hydrazone linkages. At thermodynamic equilibrium, the dominant receptor structure was an elaborate [2]-catenane consisting of two interlocked macrocyclic trimers. This complex receptor with a 100 nM affinity for acetylcholine could be isolated on a preparative scale in 67% yield.     

Isolation, characterization and antifungal activity of major constituents of the Himalayan lichen Parmelia reticulata Tayl

Antifungal activity of hexane, ethyl acetate and methanol extracts of Parmelia reticulata was evaluated against soilborne pathogenic fungi, namely, Sclerotium rolfsii, Rhizoctonia solani, R. bataticola, Fusarium udum, Pythium aphanidermatum and P. debaryanum by poisoned food technique. Maximum antifungal activity was exhibited by hexane and ethyl acetate extracts against most of the test pathogens. Secondary metabolites, namely, (±)-isousnic acid, (±)-protolichesterinic acid, atranorin, evernyl, ethyl hematommate, ethyl orsellinate, methyl hematommate (3-formyl-2,4-dihydroxy-6-methylbenzoic acid methyl ester), 2-hydroxy-4-methoxy-3,6-dimethylbenzoic acid, 1-hydroxy-3,6-dimethoxy-8-methyl-xanthen-9-one, baeomycesic acid and salazinic acid, were isolated from the above extracts and identified by 1H NMR, 13C NMR and mass spectroscopic methods. When these metabolites were tested for antifungal activity against test pathogens, maximum antifungal activity was exhibited by (±)-protolichesterinic acid against R. solani (ED50=23.09 μg mL(-1)) and P. debaryanum (ED50=16.07 μg mL(-1)) and by atranorin against S. rolfsii (ED50=39.70 μg mL(-1)). The antifungal activity of protolichesterinic acid was found to be comparable to that of hexaconazole, a commercial fungicide.     

Relative inhibition of lipid peroxidation, cyclooxygenase enzymes, and human tumor cell proliferation by natural food colors

The most abundant water soluble natural food colors are betacyanins and anthocyanins. Similarly, lycopene, bixin, beta-carotene, and chlorophyll are water insoluble colors. Pure betanin, bixin, lycopene, chlorophyll, beta-carotene, and cyanidin-3-O-glucoside were isolated from Beta vulgaris, Bixa orellana,Lycopersicum esculentum, Spinacia oleracea, Daucus carrota, and Prunus cerasus, respectively. These natural pigments, alone and in combination, were evaluated for their relative potencies against cyclooxygenase enzymes and tumor cell growth inhibition by using MCF-7 (breast), HCT-116 (colon), AGS (stomach), CNS (central nervous system), and NCI-H460 (lung) tumor cell lines. Among the colors tested, betanin, cyanidin-3-O-glucoside, lycopene, and beta-carotene inhibited lipid peroxidation. However, all pigments tested gave COX-1 and COX-2 inhibition and showed a dose-dependent growth inhibition against breast, colon, stomach, central nervous system, and lung tumor cells, respectively. The mixtures of these pigments were also evaluated for their synergistic effects and chemical interactions at various concentrations. The mixture of anthocyanin and betanin negated their efficacy in the cell growth inhibitory assay and did not enhance the COX enzyme inhibitory activity. This is the first report of a comparative evaluation and the impact on biological activities of these pigments alone and in combination.     

Canine urinary tract infections: a comparison of in vitro antimicrobial susceptibility test results and response to oral therapy with ampicillin or with trimethoprim-sulfa

In vitro susceptibility testing correctly predicted the outcome of ampicillin therapy in all 56 urinary tract infections (UTI) caused by coagulase-positive staphylococci (Staphylococcus aureus and S intermedius), in all 26 UTI caused by Proteus mirabilis, in 38 of 44 UTI caused by Escherichia coli, in 29 of 31 UTI caused by Streptococcus spp, in 8 of 10 UTI caused by Klebsiella pneumoniae, and in 16 of 20 UTI caused by other bacterial species. Thus, 173 of 187 (92.5%) isolates responded to ampicillin therapy in a manner predicted by in vitro susceptibility test results. In vitro susceptibility testing correctly predicted the outcome of therapy with trimethoprim-sulfa in 119 of 138 UTI caused by Escherichia coli, in 33 of 45 UTI caused by Klebsiella pneumoniae, in 38 of 43 UTI caused by Proteus mirabilis, in 21 of 25 UTI caused by Streptococcus spp, in 9 of 11 UTI caused by coagulase-positive staphylococci, and in 19 of 21 UTI caused by other bacterial species. Thus, 239 of 283 (84%) isolates responded to trimethoprim-sulfa therapy in a manner predicted by in vitro susceptibility test results.     

PERCUTANEOUS NEPHROPYELOCENTESIS AND ANTEGRADE URETEROGRAPHY: A FLUOROSCOPICALLY ASSISTED DIAGNOSTIC TECHNIC IN CANINE UROLOGY

Fluoroscopically assisted percutaneous nephropyelocentesis and/or antegrade ureterography was performed on six dogs. These technics were useful for obtaining renal pelvic urine specimens for culture. Percutaneous nephropyelocentesis confirmed diagnoses of pyelonephritis. Injection of a radiopaque contrast medium into the ureter or renal pelvis was useful in evaluating the ureter in those dogs in which excretory urography provided insufficient detail. No complication was associated with the technics.