Reproducible Enhancement of Fluorescence by Bimetal Mediated Surface Plasmon Coupled Emission for Highly Sensitive Quantitative Diagnosis of Double‐Stranded DNA

Plasmonic enhancement of fluorescence from SYBR Green I conjugated with a double‐stranded DNA (dsDNA) amplicon is demonstrated on polymerase chain reaction (PCR) products. Theoretical computation leads to use of the bimetallic (Au 2 nm–Ag 50 nm) surface plasmons due to larger local fields (higher quality factors) than monometallic (Ag or Au) ones at both dye excitation and emission wavelengths simultaneously, optimizing fluorescence enhancement with surface plasmon coupled emission (SPCE). Two kinds of reverse Kretschmann configurations are used, which favor, in signal‐to‐noise ratio, a fluorescence assay that uses optically dense buffer such as blood plasma. The fluorescence enhancement (12.9 fold at maximum) with remarkably high reproducibility (coefficient of variation (CV) < 1%) is experimentally demonstrated. This facilitates credible quantitation of enhanced fluorescence, however unlikely to obtain by localized surface plasmons. The plasmon‐induced optical gain of 46 dB due to SPCE‐active dye molecules is also estimated. The fluorescence enhancement technologies with PCR enables LOD of the dsDNA template concentration of ≈400 fg µL^?1 (CV < 1%), the lowest ever reported in DNA fluorescence assay to date. SPCE also reduces photobleaching significantly. These technologies can be extended for a highly reproducible and sufficiently sensitive fluorescence assay with small volumes of analytes in multiplexed diagnostics.     

Sources and leaching of manganese and iron in the Saigon River Basin, Vietnam

High concentrations of manganese and iron in the Saigon River are major problems for the water supply in Ho Chi Minh City (HCMC), Viet Nam. To identify their sources and leaching processes, we surveyed water quality along the Saigon River and ran batch leaching tests using soil and sediment samples. Two important leaching processes were identified: acidic leaching from acid sulfate soil (ASS) in the middle reaches of the river, and Mn dissolution and Fe reduction from sediments in the downstream reaches. Low pH caused the concurrent release of Fe and Mn from the ASS. In contrast, anoxia caused the release of Fe but not Mn from the sediments, whereas low pH facilitated Mn dissolution. Sediments are a more important source of Mn because of their higher Mn contents (10 times) and release rates (14 times) than those from ASS.     

Cloning and characterization of the NAD-linked glycerol-3-phosphate dehydrogenases of Trypanosoma brucei brucei and Leishmania mexicana mexicana and expression of the trypanosome enzyme in Escherichia coli

A polyclonal antiserum raised against the purified glycosomal glycerol-3-phosphate dehydrogenase of Trypanosoma brucei brucei has been used to identify the corresponding cDNA clone in a T.b. brucei expression library. This cDNA was subsequently used to obtain genomic clones containing glycerol-3-phosphate dehydrogenase genes. Two tandemly arranged genes were detected in these clones. Characterization of one of the genes showed that it codes for a polypeptide of 353 amino acids, with a molecular mass of 37,651 Da and a calculated net charge of +8. Using the T.b. brucei gene as a probe, a corresponding glycerol-3-phosphate dehydrogenase gene was also identified in a genomic library of Leishmania mexicana mexicana. The L.m. mexicana gene codes for a polypeptide of 365 amino acids, with a molecular mass of 39,140 Da and a calculated net charge of +8. The amino-acid sequences of both polypeptides are 63% identical and carry a type-1 peroxisomal targeting signal (PTS1) SKM and -SKL at their respective C-termini. Moreover, the L.m. mexicana polypeptide also carries a short N-terminal extension reminiscent of a mitochondrial transit sequence. Subcellular localisation analysis showed that in L.m. mexicana the glycerol-3-phosphate dehydrogenase activity co-fractionated both with mitochondria and with glycosomes. This is not the case in T. brucei, where the enzyme is predominantly glycosomal. The two trypanosomatid sequences resemble their prokaryotic homologues (32-36%) more than their eukaryotic counterparts (25-31%) and carry typical prokaryotic signatures. The possible reason for this prokaryotic nature of a trypanosomatid glycerol-3-phosphate dehydrogenase is discussed.     

Photoperiodic modulation of monoamines and amino-acids involved in the control of prolactin and LH secretion in the ewe: evidence for a regulation of tyrosine hydroxylase activity

The aim of this study was to analyse medico-legal autopsy rates among Norwegian citizens who died in the two northernmost counties of Norway during the 20-year period 1973-1992. Medico-legal autopsy rate was defined as the number of medico-legal autopsies divided by the total number of deaths. The rates were calculated according to year of death, manner of death, sex, age, police district and county. The material included 1539 medico-legal autopsies. In the total 20-year period 37.9% (n = 1113) of the violent deaths and 1.2% (n = 426) of the natural deaths were subjected to medico-legal autopsy. The annual rates increased gradually up to 1987. In the last 5-year period 51.7% of the violent deaths and 2.1% of the natural deaths were subjected to medico-legal autopsy. Among violent deaths in this period the medico-legal autopsy rates were: suicides 65.7%, motor vehicle traffic accidents 58.3%, falls 8.6%, and other violent deaths 77.1%. Females dying after a fall and due to natural causes were significantly less frequently than males subjected to medico-legal autopsy. These two categories of death also revealed a significant decrease in autopsy rate with increasing age (age group 0-29, 30-59 and > or = 60 years) in each 5-year period. In cases of violent death the medico-legal autopsy rate according to police district varied from 24.1% to 88.9% in the last 5-year period. In conclusion, medico-legal autopsy rates depended on manner of death, sex, age and police district, besides changes in legislation.     

Effect of the surface texturing shapes fabricated using dry etching on the extraction efficiency of vertical light-emitting diodes

On the surfaces of GaN-based light-emitting diodes (LEDs) having an n-side-up vertical electrode structure formed by the laser lift-off, various shapes of photoresist-patterned surface textures were formed by inductively coupled plasma etching and their effect on the light emission efficiencies was investigated. By the formation of various shapes of surface textures, the light output efficiency was increased from 37% to 45% compared to that without surface textures. The increase of light output efficiency was related to the increase of sidewall scattering, the decrease of reflected loss, and the decrease of cavity wall effect that occurs for the vertical LEDs by the increase of sidewall surface area.     

Brewing of glucuronic acid-enriched apple cider with enhanced antioxidant activities through the co-fermentation of yeast (Saccharomyces cerevisiae and Pichia kudriavzevii) and bacteria (Lactobacillus plantarum)

Co-fermentation using yeast (Saccharomyces cerevisiae and Pichia kudriavzevii) and the bacteria (Lactobacillus plantarum) as starters isolated from spontaneous sourdough was conducted for the brewing of glucuronic acid (GlcA)-enriched apple cider. The concentration of GlcA in the apple cider co-fermented for 14 d with commercial S. cerevisiae and L. plantarum was 37.7 ± 1.7 mg/mL while a concentration of 62.8 ± 3.1 mg/mL was recorded for fermentation with P. kudriavzevii and L. plantarum, which was higher than the corresponding single yeast fermentation. The co-fermented apple cider revealed higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of 171.67 ± 0.79 µg trolox equivalents (TE)/mL using P. kudriavzevii and L. plantarum, compared to the control (143.89 ± 7.07 µg TE/mL) just using S. cerevisiae. Thus, the co-fermentation of S. cerevisiae and L. plantarum and P. kudriavzevii and L. plantarum provided a new strategy for the development of GlcA-enriched apple cider with enhanced antioxidant capacity.     

Carob Kibble: A Bioactive‐Rich Food Ingredient

Carob (Ceratonia siliqua L.) is well known for its valuable locust bean gum obtained from the carob seeds. Separation of seeds from the pod leaves behind the carob kibble which is a good source of dietary fiber, sugars, and a range of bioactive compounds such as polyphenols and pinitol. Bioactive compounds present in carob kibble have been found to be beneficial in the control of many health problems such as diabetes, heart diseases, and colon cancer due to their antidiabetic, antioxidant, and anti‐inflammatory activities. Carob kibble has substantial potential to be used as a food ingredient. This article focuses on the composition, health benefits, and food applications of carob kibble.