A silver bullet for the treatment of depression?

The search for a rapid-acting antidepressant has been a subject of intense research interest for several decades. The article by Lucas and colleagues in this issue of Neuron provides compelling evidence from preclinical animal models that drugs acting at the serotonin 5-HT(4) receptor could finally achieve this goal. However, caution is warranted, as results from animal studies are not always predictive of therapeutic actions in humans.     

Where is the silver lining? Anxious doctors ask.

When Nova Scotia elected a Liberal government in September 1993, a wave of optimism washed over the province''s medical community. One of their own, Dr. John Savage, was now premier, and another, Dr. Ron Stewart, was minister of health. However, anticipation soon turned to anger as Stewart took aim at physician fees and hospital costs to help reduce the province''s health care budget by $62 million. Last November, relations between him and the Medical Society of Nova Scotia (MSNS) hit bottom. In an uncharacteristically political move, the society launched an ad campaign featuring slogans such as "Death by 1000 cuts" and "Uncle Sam want me. Ron Stewart doesn''t." Four months later, the health department and the MSNS called a truce, with an agreement that many physicians consider a positive step. Today the province and its doctors are speaking again, and the medical society is working hard to help define physicians'' roles in a new, regionalized health care system. But has the mood of doctors really improved? Last spring, CMAJ interviewed a cross section of Nova Scotia physicians to find out.     

Core–shell silver nanoparticles for optical labeling of cells

Silver nanoparticles have been modified with self-assembled monolayers of hydroxyl-terminated long chain thiols and encapsulated with a silica shell. The resulting core–shell nanoparticles were used as optical labels for cell analysis using flow cytometry and microscopy. The excitation of plasmon resonances in nanoparticles results in strong depolarized scattering of visible light, permitting detection at the single nanoparticle level. The nanoparticles were modified with neutravidin via epoxide–azide coupling chemistry, to which biotinylated antibodies targeting cell surface receptors were bound. The nanoparticle labels exhibited long-term stability in solutions with high salt concentrations without aggregation or silver etching. Labeled cells exhibited two orders of magnitude enhancement of the scattering intensity compared with unlabeled cells.     

mlo-based powdery mildew immunity: silver bullet or simply non-host resistance?

Durability and effectiveness against all genetic variants of a microbial species are hallmarks of so-called plant 'non-host' resistance. Highly effective immunity of monocotyledonous barley against the fungal powdery mildew pathogen, which is conferred by loss-of-function mutant alleles of the barley Mlo locus, likewise is a durable and broad-spectrum type of resistance. Although this was long considered as being a barley-specific phenomenon, recent findings indicate that mlo resistance can also occur in the distantly related dicotyledonous species Arabidopsis thaliana . Shared histological and phytopathological characteristics plus a conserved requirement for a set of genes in Arabidopsis mlo and non-host powdery mildew resistance indicate a potential common mechanism for these two seemingly distinct types of immunity.     

Optimization of PCR protocol in microsatellite analysis with silver and SYBR® stains

Here we present the optimization of PCR conditions for microsatellite analysis of coniferous trees. The use of touchdown protocol for annealing resulted in a high success rate for optimization using fewer temperature profiles. The use of SYBR^￠ Green gel stain to detect PCR products in agarose gels was more sensitive than ethidium bromide. This is valuable for determining the success of PCR reactions and estimating the amount of PCR products formed—which is crucial in determining the dilution required to produce bands of similar intensity upon silver staining of the polyacrylamide gels. The use of SYBR^￠ Gold for staining polyacrylamide gels was not satisfactory in terms of the image quality produced. However, it was comparable to silver staining in terms of sensitivity, and could possibly be used in cases where the products are present as sharp single bands. In those cases, the use of SYBR^￠ Gold gel stain would save time and money for staining polyacrylamide gels.     

Water relations in silver birch during springtime: How is sap pressurised?

• Positive sap pressures are produced in the xylem of birch trees in boreal conditions during the time between the thawing of the soil and bud break. During this period, xylem embolisms accumulated during wintertime are refilled with water. The mechanism for xylem sap pressurization and its environmental drivers are not well known.
• We measured xylem sap flow, xylem sap pressure, xylem sap osmotic concentration, xylem and whole stem diameter changes, and stem and root non‐structural carbohydrate concentrations, along with meteorological conditions at two sites in Finland during and after the sap pressurisation period.
• The diurnal dynamics of xylem sap pressure and sap flow during the sap pressurisation period varied, but were more often opposite to the diurnal pattern after bud burst, i.e. sap pressure increased and sap flow rate mostly decreased when temperature increased. Net conversion of soluble sugars to starch in the stem and roots occurred during the sap pressurisation period. Xylem sap osmotic pressure was small in comparison to total sap pressure, and it did not follow changes in environmental conditions or tree water relations.
• Based on these findings, we suggest that xylem sap pressurisation and embolism refilling occur gradually over a few weeks through water transfer from parenchyma cells to xylem vessels during daytime, and then the parenchyma are refilled mostly during nighttime by water uptake from soil. Possible drivers for water transfer from parenchyma cells to vessels are discussed. Also the functioning of thermal dissipation probes in conditions of changing stem water content is discussed.

     

Mercury from maternal “silver” tooth fillings in sheep and human breast milk

Neonatal uptake of mercury (Hg) from milk was examined in a pregnant sheep model, where radioactive mercury (Hg²⁰³)/silver tooth fillings (amalgam), were newly placed. A crossover experimental design was used in which lactating ewes nursed foster lambs. In a parallel study, the relationship between dental history and, breast milk concentration of Hg was also examined in 33 lactating women. Results from the animal studies showed that, during pregnancy, a primary fetal site of amalgam Hg concentration is the liver, and, after delivery, the neonatal lamb kidney receives additional amalgam Hg from mother's milk. In lactating women with aged, amalgam fillings, increased Hg excretion in breast milk and urine correlated with the number of fillings or Hg vapor concentration levels in mouth air. It was concluded that Hg originating from maternal amalgam tooth fillings transfers across the placenta to the fetus, across the mammary gland into milk ingested by the newborn, and ultimately into neonatal body tissues. Comparisons are made to the U.S. minimal risk level recently established for adult Hg exposure. These findings suggest, that placement and removal of “silver” tooth fillings in pregnant and lactating humans will subject the fetus and neonate to unnecessary risk of Hg exposure.     

Two new N-heterocyclic carbene silver(I) complexes with the π–π stacking interactions

The precursors bis[N-(alkyl)benzimidazoliumylmethyl]durene halide (1a: alkyl = C₂H₅, halide = Br^?; 1b: alkyl = n-C₄H₉, halide = Cl^?; durene = 1,2,4,5-tetramethylbenzene) and their two new NHC silver(I) complexes [Durene(CH₂BimyEtAgBr)₂] (2a) and [Durene(CH₂BimyⁿBuAgCl)₂] (2b) (Bimy = benzimidazol-2-ylidene) have been prepared and characterized. In the crystal structures of 2a and 2b the aromatic π–π stacking interactions are observed.     

No silver bullet? Snow leopard prey selection in Mt. Kangchenjunga,Nepal

In this study, we investigated the impact of domestic and wild prey availability on snow leopard prey preference in the Kangchenjunga Conservation Area of eastern Nepal—a region where small domestic livestock are absent and small wild ungulate prey are present. We took a comprehensive approach that combined fecal genetic sampling, macro‐ and microscopic analyses of snow leopard diets, and direct observation of blue sheep and livestock in the KCA. Out of the collected 88 putative snow leopard scat samples from 140 transects (290 km) in 27 (4 × 4 km²) sampling grid cells, 73 (83%) were confirmed to be from snow leopard. The genetic analysis accounted for 19 individual snow leopards (10 males and 9 females), with a mean population size estimate of 24 (95% CI: 19–29) and an average density of 3.9 snow leopards/100 km² within 609 km². The total available prey biomass of blue sheep and yak was estimated at 355,236 kg (505 kg yak/km² and 78 kg blue sheep/km²). From the available prey biomass, we estimated snow leopards consumed 7% annually, which comprised wild prey (49%), domestic livestock (45%), and 6% unidentified items. The estimated 47,736 kg blue sheep biomass gives a snow leopard‐to‐blue sheep ratio of 1:59 on a weight basis. The high preference of snow leopard to domestic livestock appears to be influenced by a much smaller available biomass of wild prey than in other regions of Nepal (e.g., 78 kg/km² in the KCA compared with a range of 200–300 kg/km² in other regions of Nepal). Along with livestock insurance scheme improvement, there needs to be a focus on improved livestock guarding, predator‐proof corrals as well as engaging and educating local people to be citizen scientists on the importance of snow leopard conservation, involving them in long‐term monitoring programs and promotion of ecotourism.     

Multispectroscopic studies on the molecular interactions between bovine γ-globulin and borohydride-capped silver nanoparticles

Interactions between bovine γ-globulin (BGG) and borohydride-capped silver nanoparticles (BAgNPs) were studied using dynamic light scattering (DLS) and spectroscopic techniques such as UV–vis spectroscopy, fluorescence, and circular dichroism. The results were compared with earlier reported interactions between γ-globulin and citrate-coated AgNPs (CAgNPs). BAgNPs were synthesized and characterized. Irrespective of the coating on AgNPs, nanoparticles had formed ground-state complexes with the protein. CAgNPs, as well as BAgNPs had caused static quenching of tryptophan (Trp) fluorescence of the protein. The change in the capping agent from citrate to borohydride weakened the binding of nanoparticles with the protein. But the same change in capping agent had increased the fluorescence quenching efficiency of AgNPs. Hydrogen bonding and van der Waals interactions were involved in BGG–BAgNPs complex similar to the CAgNPs complex with γ-globulin. Polarity of the Trp microenvironment in BGG was not altered using BAgNPs as opposed to CAgNPs, as supported using synchronous and three-dimensional fluorescence. Resonance light scattering experiments also suggested nano-bio conjugation. Far-UV and near-UV circular dichroism (CD) spectra respectively pointed towards changes in the secondary and tertiary structure of BGG by BAgNPs, which was not observed for CAgNPs.     

Simultaneous determination of the amounts of metallic and “reducible” silver in histologic specimens

Summary Acids and weak complexing agents (pK<8) are not able to remove, without leaving a residue, silver bound to biological tissues by ionic or complex bonds (reducible silver), whereas, strong complexing agents (pK>8) can also partially or completely dissolve metallic silver formed under the influence of reducing groups in the tissue. For this reason, the chemical nature of the silver contained in tissue sections, be it metallic or reducible, must not be determined on the basis of solubility tests; moreover, the amount of neither of the two above fractions can be determined by removing the other with any kind of washing. Using radioactive impregnating baths, radioactive silver bound to the tissue as reducible silver can be replaced in a quantitative manner with inactive silver ions by means of a one-hour incubation in 1% inactive silver nitrate dissolved in 10% acetic acid, but the radioactive silver existing in reduced (atomic) state will be left unaffected. Consequently, radioactivity remaining in the tissue after the above treatment represents metallic silver. The amount of reducible silver can be calculated by subtracting that of the metallic silver from the total silver content of the sections.     

Arginine methylation: the promise of a ‘silver bullet’ for brain tumours?

Despite intense research efforts, our pharmaceutical repertoire against high-grade brain tumours has not been able to increase patient survival for a decade and life expectancy remains at less than 16 months after diagnosis, on average. Inhibitors of protein arginine methyltransferases (PRMTs) have been developed and investigated over the past 15 years and have now entered oncology clinical trials, including for brain tumours. This review collates recent advances in the understanding of the role of PRMTs and arginine methylation in brain tumours. We provide an up-to-date literature review on the mechanisms for PRMT regulation. These include endogenous modulators such as alternative splicing, miRNA, post-translational modifications and PRMT–protein interactions, and synthetic inhibitors. We discuss the relevance of PRMTs in brain tumours with a particular focus on PRMT1, -2, -5 and -8. Finally, we include a future perspective where we discuss possible routes for further research on arginine methylation and on the use of PRMT inhibitors in the context of brain tumours.

     

Immobilization of horseradish peroxidase on β-cyclodextrin-capped silver nanoparticles: Its future aspects in biosensor application

This study aimed to work out a simple and high-yield procedure for the immobilization of horseradish peroxidase on silver nanoparticle. Ultraviolet–visible (UV-vis) and Fourier-transform infrared spectroscopy and transmission electron microscopy were used to characterize silver nanoparticles. Horseradish peroxidase was immobilized on β-cyclodextrin-capped silver nanoparticles via glutaraldehyde cross-linking. Single-cell gel electrophoresis (Comet assay) was also performed to confirm the genotoxicity of silver nanoparticles. To decrease toxicity, silver nanoparticles were capped with β-cyclodextrin. A comparative stability study of soluble and immobilized enzyme preparations was investigated against pH, temperature, and chaotropic agent, urea. The results showed that the cross-linked peroxidase was significantly more stable as compared to the soluble counterpart. The immobilized enzyme exhibited stable enzyme activities after repeated uses.     

Antibacterial efficacy and cytotoxicity of low intensity direct current activated silver–titanium implant system prototype

Silver-based devices activated by electric current are of interest in biomedicine because of their broad-spectrum antimicrobial activity. This study investigates the in vitro antibacterial efficacy and cytotoxicity of a low intensity direct current (LIDC)-activated silver–titanium implant system prototype designed for localized generation and delivery of silver ions at the implantation site. First, the antibacterial efficacy of the system was assessed against methicillin-resistant Staphylococcus aureus (MRSA) over 48 h at current levels of 3 and 6 µA in Mueller–Hinton broth. The cytotoxicity of the system was then evaluated over 48 h in two phases using an in vitro model with in which the activated electrodes were suspended in growth medium in a cell-seeded tissue culture plate. In phase-1, the system was tested on human osteosarcoma (MG-63) cell line and compared to titanium controls. In phase-2, the cytotoxicity characteristics were validated with normal human diploid osteoblast cells. The LIDC-activated system demonstrated high antimicrobial efficacy against MRSA, but was also toxic to human cells immediately surrounding the electrodes. The statistical analysis showed that the cytotoxicity was a result of the presence of silver, and the electric activation did not make it worse.     

Is alcoholic acidic silver nitrate reagent really specific for the histochemical localization of ascorbic acid?

Summary The histochemical localization of ascorbic acid in plant tissues with the alcoholic acidic silver nitrate reagent is shown here to be not specific for ascorbic acid, since some of the polyphenolic substances, including flavonoids, which are known to be widely distributed in plant tissues, are also able to reduce the acidic alcoholic silver nitrate reagent at low temperature (0–4°C) and at pH 2 to 2.5 in dark. This method may perhaps be used for animal tissues where flavonoid pigments do not occur in such large quantities as they do in plants. I therefore, come to the inevitable conclusion that the use of alcoholic acidic silver nitrate reagent in localizing ascorbic acid in plant tissues may be highly misleading.     

Identification of genes required for gold and silver tolerance in Burkholderia cenocepacia H111 by transposon sequencing

Members of the genus Burkholderia show remarkable abilities to adapt to a wide range of environmental conditions and is frequently isolated from soils contaminated with heavy metals. In this study, we used a transposon sequencing approach to identify 138 and 164 genes that provide a benefit for growth of the opportunistic pathogen Burkholderia cenocepacia H111 in the presence of silver and gold ions respectively. The data suggest that arginine metabolism and citrate biosynthesis are important for silver tolerance, while components of an ABC transporter (BCAL0307-BCAL0308) and de novo cysteine biosynthesis are required for tolerance to gold ions. We show that determinants that affect tolerance to both metal ions include the two-component systems BCAL0497/99 and BCAL2830/31 and genes that are involved in maintaining the integrity of the cell envelope, suggesting that membrane proteins represent important targets of silver and gold ions. Furthermore, we show that that the P-type ATPase CadA (BCAL0055), which confers tolerance to cadmium contributes to silver but not gold tolerance. Our results may be useful for improving the antibacterial effect of silver and gold ions to combat drug-resistant pathogens.     

Modified protocols for rapid carrot genomic DNA extraction and AFLP™ analysis using silver stain or radioisotopes

AFLP^™ is one of the most frequently used techniques for identification of molecular markers. We have modified the procedures for genomic DNA extraction, AFLP product generation and silver staining in order to speed up analyses and screen large numbers of plant samples. Using this protocol, we were able to achieve an 82% reduction of costs without compromising the reliability and quality of data gathered.     

Effects of invasive cane toads on Australian mosquitoes: Does the dark cloud have a silver lining?

Research on the ecological impacts of invasive organisms typically looks only for negative impacts, ignoring the possibility that the wider community might see benefits in some of these effects. To truly understand the impact of invasive species, we need to look as broadly as possible, and incorporate studies on a diversity of variables. The spread of the South American cane toad (Bufo marinus) through tropical Australia is widely viewed as an ecological catastrophe, but anecdotal reports suggest that the invasion of toads may reduce the numbers of mosquitoes (and thus, potentially, the risk they pose to human health). We conducted experiments to determine whether the presence of toad tadpoles affects survival rates, adult body sizes and/or rates of oviposition of four species of disease-carrying mosquitoes. In the laboratory, the presence of toad tadpoles significantly reduced the sizes of adult mosquitoes at emergence, and also reduced survival rates of the larvae of one mosquito species. In field trials, mosquitoes were less likely to oviposit in waterbodies containing toad tadpoles. Accordingly, these data suggest (but do not prove) that toad invasion may reduce mosquito abundance. More generally, any overall evaluation of the impact of an invasive species needs to consider possible benefits (e.g. to human health) as well as negative effects (e.g. to native species). Both types of information are essential to inform community decisions about the management of feral taxa such as the cane toad in Australia.