In vitro efficacy of nisin Z against Candida albicans adhesion and transition following contact with normal human gingival cells

Aim:  To investigate the nisin Z innocuity using normal human gingival fibroblast and epithelial cell cultures, and its synergistic effect with these gingival cells against Candida albicans adhesion and transition from blastospore to hyphal form.
Methods and Results:  Cells were cultured to 80% confluence and infected with C. albicans in the absence or presence of various concentrations of nisin Z. Our results indicate that only high concentrations of nisin Z promoted gingival cell detachment and differentiation. Determination of the LD₅₀ showed that the fibroblasts were able to tolerate up to 80  μ g ml⁻¹ for 24 h, dropping thereafter to 62  μ g ml⁻¹ after 72 h of contact, compared to 160  μ g ml⁻¹ after 24 h, and 80  μ g ml⁻¹ after 72 h recorded by the gingival epithelial cells which displayed a greater resistance to nisin Z. The use of nisin Z even at low concentration (25  μ g ml⁻¹) at appropriate concentrations with gingival cells significantly reduced C. albicans adhesion to gingival monolayer cultures and inhibited the yeast's transition.
Conclusion:  These findings show that when used at non-toxic levels for human cells, nisin Z can be effective against C. albicans adhesion and transition and may synergistically interact with gingival cells for an efficient resistance against C. albicans .
Significance and Impact of the Study:  This study suggests the potential usefulness of nisin Z as an antifungal agent, when used in an appropriate range.     

Microcystin-producing blooms of Anabaenopsis arnoldi in a potable mountain lake in Saudi Arabia

This study reports for the first time the presence of Anabaenopsis arnoldi blooms in Saudi freshwaters. This species has been investigated with high cell densities (3.8 × 10³–264 × 10³ cells mL⁻¹) during June–November 2007 in Tendaha Lake, one of the major freshwater sources in Saudi Arabia. High temperature and conductivity, and a high concentration of phosphate, and low nitrate concentrations may have contributed to the formation of these blooms. The blooms were found to produce microcystins (MCYSTs) at concentrations up to 364 μg g⁻¹ dry weight as detected by an enzyme-linked immunosorbent assay. MCYSTs were also detected in the raw and treated water of the lake at concentrations (1.6–8.3 and 0.33–1.6 μg L⁻¹, respectively) exceeding the World Health Organization guideline level of 1 μg L⁻¹ for these toxins. HPLC analysis revealed that the extracts of A. arnoldi blooms contained MCYST-RR, -YR and two unidentified MCYSTs, but a pure culture of A. arnoldi isolated from Tendaha Lake during the present study produced MCYST-RR and –YR only. This is the first study to report MCYST production by A. arnoldi . Therefore, this cyanobacterium should be taken into consideration during monitoring of toxic cyanobacterial blooms in drinking and recreational water sources in the world, particularly arid and semi-arid countries including Saudi Arabia.     

The synergistic activity of triclosan and ciprofloxacin on biofilms of Salmonella Typhimurium

Triclosan is a biocide whose wide use has raised a debate about the potential benefits vs. hazards of the incorporation of antimicrobials in consumer products. The purpose of the present study was to determine whether exposure of biofilms of Salmonella enterica serovar Typhimurium to triclosan influences the tolerance of the bacteria towards antibiotics such as ciprofloxacin and vice versa. A synergistic antibiofilm activity was observed when the biofilms were treated with triclosan before or together with ciprofloxacin, and an additive activity was observed with planktonic cells. For example 500 μg mL⁻¹ triclosan and 500 μg mL⁻¹ ciprofloxacin reduced the number of viable cells in the biofilm by 1.6 and 0.5 log, respectively. However, the sequential treatment of 500 μg mL⁻¹ triclosan followed by ciprofloxacin resulted in 4.8 log reduction. Combination indexes (CI) for biofilms treated with triclosan followed by ciprofloxacin were 0.7, 0.32 and 0.25 for reduction of 90%, 99% and 99.9%, respectively, indicating a synergism. For planktonic cells, CIs were 1±0.1, indicating an additive effect. Therefore, it was suggested that triclosan weakens the ability of biofilm-associated cells to survive exposure to ciprofloxacin in the biofilm, probably by improving the permeability or the activity of ciprofloxacin.     

Development and characterization of a lux-modified 2,4-dichlorophenol-degrading Burkholderia sp. RASC

lux -marked biosensors for assessing the toxicity and bioremediation potential of polluted environments may complement traditional chemical techniques. lux CDABE genes were introduced into the chromosome of the 2,4-dichlorophenol (2,4-DCP)-mineralizing bacterium, Burkholderia sp. RASC c2, by biparental mating using the Tn 4431 system. Experiments revealed that light output was constitutive and related to cell biomass concentration during exponential growth. The transposon insertion was stable and did not interrupt 2,4-DCP-degradative genes, and expression of lux CDABE did not constitute a metabolic burden to the cell. A bioluminescence response was detectable at sublethal 2,4-DCP concentrations: at < 10.26 μg ml⁻¹, bioluminescence was stimulated (e.g. 218% of control), but at concentrations > 60 μg ml⁻¹ it declined to < 1%. Investigating the effect of [14C]-2,4-DCP concentration on the evolution of ¹⁴CO₂ revealed that, for initial concentrations of 2.5–25 μg ml⁻¹, ≈55% of the added ¹⁴C was mineralized after 24 h compared with < 1% at 50 and 100 μg ml⁻¹. Inhibition of 2,4-DCP mineralization between 25 and 50 μg ml⁻¹ corresponded well to the EC₅₀ value (33.83 μg ml⁻¹) obtained from bioluminescence inhibition studies. lux -marked RASC c2 may therefore be used as a functionally (i.e. 2,4-DCP degrader) and environmentally relevant biosensor of toxicity and biodegradation inhibition.     

Constitutive mineralization of low concentrations of the herbicide linuron by a Variovorax sp. strain

The mineralization of the herbicide linuron at concentrations of μg and mg L⁻¹ was studied in liquid batch experiments with Variovorax sp. strain SRS16. The strain was highly efficient at mineralizing a range of linuron concentrations (0.002–10 mg L⁻¹) with 20–60% of the added ¹⁴C-ring-labeled linuron metabolized to ¹⁴CO₂ within hours to days depending on the initial linuron concentration and incubation period. At mg L⁻¹ linuron concentrations the mineralization activity by SRS16 was inducible and a shift to constitutive mineralization activity was apparent with a reduction in the linuron concentration to μg L⁻¹ levels. This study revealed that strain SRS16 is a promising candidate for bioaugmentation of water or soil resources contaminated with low linuron concentrations.     

More effective induction of spawning with long-acting GnRH agonist in the shi drum, Umbrina cirrosa L. (Sciaenidae, Teleostei), a valuable candidate for Mediterranean mariculture

Three experiments of spawning induction in shi drum, Umbrina cirrosa L., were performed in six different commercial Italian hatcheries from May to August (water temperatures: 19–29 °C; salinity: 21–37 p.p.t.). In the first experiment, 119 females (1–4.7 kg), subdivided into 29 lots, were injected with a single dose (2, 5, 8, 10, 15 and 20  μ g kg⁻¹ body weight) of short-acting gonadotropin- releasing hormone agonist (GnRHa-S), des-Gly10,[D-Ala6]-LH-RH ethylamide. In the other two experiments, 85 females (0.7–5.8 kg), subdivided into 22 and four lots, were treated with one (40 or 80  μ g kg⁻¹) or three doses (40  μ g kg⁻¹) of long-acting GnRHa (GnRHa-L), respectively. GnRHa-S stimulated spawning in 69% of the 29 treated lots; the number of eggs laid reached a maximum of 130 000 and a weighted mean of 29 200 total eggs kg⁻¹. GnRHa-L elicited a spawning response in 95% of the 22 one-dose treated lots; the number of laid eggs was higher than with GnRHa-S, reaching a maximum of 213 100 and a weighted mean of 59 400 total eggs kg⁻¹. The yield of developing embryos in 67% of the single GnRHa-L treatments was higher (sometimes up to three times) than with GnRHa-S. Triple treatments of the four lots of females with GnRHa-L always resulted in spawning responses; the best result corresponded to a number of total laid eggs of 358 900 eggs kg⁻¹ with a yield of 177 300 developing embryos.     

Toll-like receptors 2 and 4 mediate Abeta(1-42) activation of the innate immune response in a human monocytic cell line

The primary molecules for mediating the innate immune response are the Toll-like family of receptors (TLRs). Recent work has established that amyloid-beta (Aβ) fibrils, the primary components of senile plaques in Alzheimer's disease (AD), can interact with the TLR2/4 accessory protein CD14. Using antibody neutralization assays and tumor necrosis factor alpha release in the human monocytic THP-1 cell line, we determined that both TLR2 and TLR4 mediated an inflammatory response to aggregated Aβ(1–42). This was in contrast to exclusive TLR ligands lipopolysaccharide (LPS) (TLR4) and tripalmitoyl cysteinyl seryl tetralysine (Pam₃CSK₄) (TLR2). Atomic force microscopy imaging showed a fibrillar morphology for the proinflammatory Aβ(1–42) species. Pre-treatment of the cells with 10 μg/mL of a TLR2-specific antibody blocked ∼50% of the cell response to fibrillar Aβ(1–42), completely blocked the Pam₃CSK₄ response, and had no effect on the LPS-induced response. A TLR4-specific antibody (10 μg/mL) blocked ∼35% of the cell response to fibrillar Aβ(1–42), completely blocked the LPS response, and had no effect on the Pam₃CSK₄ response. Polymyxin B abolished the LPS response with no effect on Aβ(1–42) ruling out bacterial contamination of the Aβ samples. Combination antibody pre-treatments indicated that neutralization of TLR2, TLR4, and CD14 together was much more effective at blocking the Aβ(1–42) response than the antibodies used alone. These data demonstrate that fibrillar Aβ(1–42) can trigger the innate immune response and that both TLR2 and TLR4 mediate Aβ-induced tumor necrosis factor alpha production in a human monocytic cell line.     

Quantitative method for pheromone delivery in studies of sensory adaptation of moth antennae

Abstract.  A pheromone sprayer and an electroantennogram (EAG) are used to study sensory adaptation in the antennae of male obliquebanded leafrollers, Choristoneura rosaceana and oriental fruit moths, Grapholita molesta , to the main pheromone compounds ( Z )-11-tetradecen-1-yl acetate ( Z 11-14:Ac) and ( Z )-8-dodecen-1-yl acetate ( Z 8-12:Ac), respectively. The atomization of 0.125, 0.25, 0.5 or 1 μL ethanol min⁻¹ into the EAG air delivery tube at an airflow rate of 2 L min⁻¹, with resultant concentrations of 6.25, 12.5, 25 or 50 × 10⁻⁵μL ethanol mL air⁻¹, respectively, does not affect the EAG response of C. rosaceana or C. molesta after a 30-min exposure period. The atomization of 0.125 μL min⁻¹ of a solution of 8 mg Z 11-14:Ac mL⁻¹ ethanol into the EAG air delivery tube at an airflow rate of 2 L min⁻¹, with a resultant concentration of 0.5 ng pheromone mL⁻¹ air, reduces the EAG response of C. rosaceana by approximately 70% after a 15-min exposure period. An additional 15 min of exposure to pheromone does not result in increased sensory adaptation. Antennae recover 32% of the lost responsiveness when exposed to pheromone-free air for 15 min. The atomization of 0.125 μL min⁻¹ of a solution of 8 mg Z 8-12:Ac mL⁻¹ ethanol into the EAG air delivery tube at an airflow rate of 2 L min⁻¹, with a resultant concentration of 0.5 ng pheromone mL⁻¹ air, reduces the EAG response of C. molesta antenna by approximately 80% after a 15- or 30-min exposure period. The antennae of this species do not recover responsiveness when exposed to pheromone-free air for 15 min.     

Investigating the effects of positive charge and hydrophobicity on the cell selectivity, mechanism of action and anti-inflammatory activity of a Trp-rich antimicrobial peptide indolicidin

To investigate the effects of positive charge and hydrophobicity on the cell selectivity, mechanism of action and anti-inflammatory activity of a Trp-rich antimicrobial peptide indolicidin (IN), a series of IN analogs with Trp→Lys substitution were synthesized. All IN analogs displayed an approximately 7- to 18-fold higher cell selectivity, compared with IN. IN, IN-1 and IN-2 depolarized (50−90%) the cytoplasmic membrane potential of Staphylococcus aureus close to minimal inhibitory concentration (5–10 μg mL⁻¹). However, other IN analogs (IN-3 and IN-4) displayed very low ability in membrane depolarization even at 40 μg mL⁻¹. Confocal laser-scanning microscopy revealed that IN-3 and IN-4 penetrated the Escherichia coli cell membrane, whereas IN, IN-1 and IN-2 did not enter the cell membrane. In the gel retardation assay, IN-3 and IN-4 bound more strongly to DNA compared with IN, IN-1 and IN-2. These findings suggest that the mechanism of antimicrobial action of IN-3 and IN-4 may be involved in the inhibition of intracellular functions via interference with DNA/RNA synthesis. Unlike IN, all IN analogs did not inhibit nitric oxide production or inducible nitric oxide synthase mRNA expression in lipopolysaccharide-stimulated mouse macrophage RAW264.7 cells, indicating that the hydrophobicity of IN is more important for anti-inflammatory activity in lipopolysaccharide-treated macrophage cells than the positive charge.     

Breakdown of tussock grass in streams along a gradient of agricultural development in New Zealand

Summary 1. We measured the breakdown rate of tussock grass in 12 New Zealand streams in catchments that provided a gradient of agricultural development. We also examined the microbial and invertebrate communities associated with decomposing tussock litter.
2. Pristine streams in the study had low concentrations of dissolved inorganic nitrogen (<10  μ g L⁻¹) and dissolved reactive phosphate (<3  μ g L⁻¹), whereas streams in the most developed catchments had high concentrations of nitrate (>2500  μ g L⁻¹) and phosphate (35  μ g L⁻¹), as well as greater amounts of suspended sediment and fine sediment covering the streambed.
3. Breakdown rate and microbial respiration were significantly related across the sites, and both were positively related to concentrations of nitrate and phosphate. Fungal biomass, measured as ergosterol, was positively related to microbial respiration and was also higher at sites with higher concentrations of nutrients. Total and shredding invertebrates were most abundant at the sites with high nutrient concentrations, but abundance of shredding invertebrates was not significantly related to breakdown rate. Amphipods were the most common shredding invertebrate at most sites, but probably did not contribute greatly to high rates of breakdown in streams in agricultural catchments.
4. With the exception of one site, nutrients from agricultural development appeared to have larger positive effects on litter breakdown than negative effects from sedimentation. Litter breakdown can serve as a functional measure of ecosystem health in streams, but caution should be exercised when a stress, such as land use, can have both positive (nutrients) and negative (sedimentation) effects.     

Influence of phosphorus on biofilm formation in model drinking water distribution systems

Aims:  This study investigated the effects of phosphorus on biofilm formation via annular reactor systems in terms of biofilm cell growth, exopolysaccharide (EPS) production, biofilm structure and cell metabolic potential.
Methods and Results:  Drinking water biofilms were developed in annular reactors with supplement of carbon and different levels of phosphorus. The biofilm formation was monitored over a period of 30 days. Biofilm related parameters were examined by various methods, which included heterotrophic plate count, total carbohydrate content, confocal laser scanning microscopy and GN2 microplate assay. Our results showed that phosphorus addition can promote the biofilm cell growth (cell count increased about 1 log with addition of 30 and 300 μg l⁻¹ of phosphorus). However, the addition of 30 and 300 μg l⁻¹ of phosphorus caused 81% and 77% decrease in EPS production, respectively. The results of biofilm structure analysis showed that the addition of 30 and 300 μg l⁻¹ of phosphorus can induce thicker and less homogeneous biofilms with more biomass. Furthermore, the addition of 30 and 300 μg l⁻¹ of phosphorus dramatically increased the biofilm cell metabolic potential. The addition of 3 μg l⁻¹ of phosphorus was found to have minor effects on the parameters examined.
Conclusions:  The results indicate phosphorus addition to drinking water distribution system (DWDS) has a complicated effect on the biofilm formation.
Significance and Impact of the Study:  As the addition of phosphorus at certain levels can affect the biofilm growth in DWDS, care should be taken when phosphate-based corrosion inhibitors are used in the DWDS.     

Early life stage toxicity of extracts from the African fish poison plants Tephrosia vogelii Hook. f. and Asystasia vogeliana Benth. on zebrafish embryos

Embryos of Danio rerio are highly susceptible to extracts of the plants Tephrosia vogelii and Asystasia vogeliana . The concentration of the dried extracts at which 50% of the embryos were affected (EC₅₀) after 24 h exposure were 320 and 572 μg l⁻¹, respectively; corresponding 50% mortality (LC₅₀) values after 48 h exposure were 493 and 869 μg l⁻¹. Results indicate that the use of these ichthyotoxic plants might have a severe impact on the survival of fish larvae in the field.     

The mode of antistaphylococcal action of Eleutherine americana

The anti-methicillin-resistant Staphylococcus aureus (anti-MRSA) activity and the possible mechanism of action of a crude extract from red bulbs of Eleutherine americana Merr. were investigated. The crude ethanolic extract from E. americana produced minimum inhibitory concentrations (MICs) of 62.5–1000 and 250 μg mL⁻¹ against MRSA isolates and the reference strains, respectively. Treatment of S. aureus ATCC 27664 with a crude extract at 2MIC reduced the inoculum size by 5 log at 24 h compared with the control. The combined effect of the extract and 7.5% NaCl on the enterotoxin-producing ATCC strain resulted in no detection of organisms within 24 h compared with the control. The release of cell materials after extract treatment was determined by measuring OD_260 nm, the treatment resulted in cytoplasmic leakage. Determination of OD_620 nm showed that the extract did not cause gross cell wall damage. However, observation of S. aureus cells under an electron microscope after treatment with 2MIC and 4MIC of the crude extract revealed that the extract caused damage to membrane morphology. A knowledge of the mechanism of action of the E. americana extract may offer useful hints in the search for novel antibacterial substance.     

In Human Fetal Astrocytes Exposure to Interleukin-1β Stimulates Acquisition of the GD3+ Phenotype and Inhibits Cell Division

Abstract: In human astrocyte cultures established from second-trimester fetal brain tissue, ∼5–10% of total astrocyte population in unstimulated cultures were GD3⁺/glial fibrillary acidic protein (GFAP)⁺. The GD3⁺ cells were always GFAP⁺ and grew as flat, highly spread cells but changed to process-bearing cells after interleukin-1β (IL-1β) stimulation. It is interesting that IL-1β, a known mitogen for rat astrocytes, suppressed human fetal astrocyte proliferation as determined by [³H]thymidine incorporation, bromodeoxyuridine (BrdU) labeling, and cell counting. The GD3⁺ population, however, consistently increased in absolute number after IL-1β stimulation, in a dose- and time-dependent manner. The IL-1β-mediated increase in number of GD3⁺ astrocytes was independent of initial cell density or serum concentration. By flow cytometry, IL-1β enhanced both the mean fluorescence intensity and the percentage of GD3⁺ cells. To investigate whether the increase in GD3⁺ astrocyte cell number was due to proliferation of preexisting GD3⁺ astrocytes or due to conversion of GD3⁻ to GD3⁺ cells, we performed BrdU/GD3 double immunocytochemistry. BrdU/GD3 double-positive cells were extremely rare in both control and IL-1β-stimulated cultures. Moreover, an increase in number of GD3⁺ astrocytes was still observed in control and IL-1β-stimulated cultures where GD3⁺ cells had been initially eliminated by cell sorting. These results indicate that GD3⁺ astrocytes in human fetal culture may represent a postmitotic, differentiated, distinct phenotype.     

Response of Listeria monocytogenes to liquid smoke

Aims:  To investigate the effect of liquid smoke on growth, survival, proteomic pattern and haemolytic potential of Listeria monocytogenes.
Methods and Results:  Growth and survival curves were recorded in brain–heart infusion broth supplemented with three concentrations of liquid smoke. L. monocytogenes growth was inhibited in the presence of 15 μg ml⁻¹ phenol while a rapid decrease in cell viability occurred in the presence of 30 μg ml⁻¹ phenol. The proteome of L. monocytogenes cytosoluble proteins was slightly modified after 2-h incubation with 30 μg ml⁻¹ phenol but no protein already characterized in response to other known stresses was induced, except the protease ClpP. Liquid smoke inhibited the haemolytic potential without affecting hly gene expression, showing a potential inhibition of protein activity or stability.
Conclusions:  The presence of liquid smoke in a rich medium strongly affected growth and survival of L. monocytogenes . Brief smoke stress affected the metabolic pathways and inhibited the haemolytic activity of L. monocytogenes .
Significance and Impact of Study:  This study is a first step in the investigation of the influence of a smoked product on L. monocytogenes strains.     

Biochemical characterization of a β-fructofuranosidase from Rhodotorula dairenensis with transfructosylating activity

An extracellular β-fructofuranosidase from the yeast Rhodotorula dairenensis was characterized biochemically. The enzyme molecular mass was estimated to be 680 kDa by analytical gel filtration and 172 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, of which the N -linked carbohydrate accounts for 16% of the total mass. It displays optimum activity at pH 5 and 55–60 °C. The enzyme shows broad substrate specificity, hydrolyzing sucrose, 1-kestose, nystose, leucrose, raffinose and inulin. Although the main reaction catalyzed by this enzyme is sucrose hydrolysis, it also exhibits transfructosylating activity that, unlike other microbial β-fructofuranosidases, produces a varied type of prebiotic fructooligosaccharides containing β-(2→1)- and β-(2→6)-linked fructose oligomers. The maximum concentration of fructooligosaccharides was reached at 75% sucrose conversion and it was 87.9 g L⁻¹. The 17.0% (w/w) referred to the total amount of sugars in the reaction mixture. At this point, the amounts of 6-kestose, neokestose, 1-kestose and tetrasaccharides were 68.9, 10.6, 2.6 and 12.7 g L⁻¹, respectively.     

Bactericidal activity of catechin-copper (II) complexes on Escherichia coli ATCC11775 in the absence of hydrogen peroxide

Washed Escherichia coli ATCC11775 cells were killed by (–)-epigallocatechin (EGC) in the presence of a non- lethal concentration of Cu²⁺ (1 μmol l⁻¹) without additional H₂O₂, but not by (–)-epicatechin (EC). EGC alone (< 0·1 mmol l⁻¹) did not reduce the viability of the cells. The survival curve obtained in the presence of EGC and Cu²⁺ was similar to that obtained in the presence of (–)-adrenaline (EN) and Cu²⁺.     

Acrebol, a novel toxic peptaibol produced by an Acremonium exuviarum indoor isolate

Aims:  To identify a toxin and its producer isolated from woody material in a building where the occupants experienced serious ill health symptoms.
Methods and Results:  Hyphal extracts of an indoor fungus, identified as the cycloheximide-tolerant species Acremonium exuviarum , inhibited motility of boar spermatozoa (EC₅₀ 5 ± 2 μg of crude solids ml⁻¹) and caused cytolysis of murine neuroblastoma cells (MNA) and feline fetal lung cells (FL). The responsible substances were purified and identified as two structurally similar, heat-stable, novel, toxic peptaibols, 1726 Da and 1740 Da, respectively, with amino acid sequences of Acetyl-Phe-Iva/Val-Gln-Aib-Ile-Thr-Leu-Aib-Pro-Aib-Gln-Pro-Aib-(X-X-X)-SerOH and Acetyl-Phe-Iva/Val-Gln-Aib-Ile-Thr-Leu-Val-Pro-Aib-Gln-Pro-Aib-(X-X-X)-SerOH. Purified acrebol inhibited motility of boar sperm, depleted ATP half-content in 1 day (EC₅₀ of 0·1 μg ml⁻¹, 60 nmol l⁻¹) depolarised the mitochondria after 2 days, but did not affect the cellular content in NADH. This indicates mitochondrial toxicity. Plate-grown biomass of A. exuviarum BMB4 contained 0·1–1% (w/w) of acrebol, depending on the culture medium.
Conclusions:  Acrebol paralysed the energy generation of mammalian cells suggesting that mitochondria were its target of action.
Significance and Impact of the Study:  Acremonium exuviarum, as an indoor fungus, is potentially hazardous to health because of the toxic peptaibols that it produces.     

Rapid Communication: Ca2+ Channel Blockers Attenuate β-Amyloid Peptide Toxicity to Cortical Neurons in Culture

Abstract: Deposit of β-amyloid protein (Aβ) in Alzheimer's disease brain may contribute to the associated neurodegeneration. We have studied the neurotoxicity of Aβ in primary cultures of murine cortical neurons, with the aim of identifying pharmacologic ways of attenuating the injury. Exposure of cultures to Aβ (25–35 fragment; 3–25 4mU M ) generally triggers slow, concentration-dependent neurodegeneration (over 24–72 h). With submaximal Aβ- (25–35) exposure (10 μ M ), substantial (>40% within 48 h) degeneration often occurs and is markedly attenuated by the presence of the Ca²⁺ channel blockers nimodipine (1–20 μ M ) and Co²⁺ (100 μ M ) during the Aβ exposure. However, Aβ neurotoxicity is not affected by the presence of glutamate receptor antagonists. We suggest that Ca²⁺ influx through voltage-gated Ca²⁺ channels may contribute to Aβ-induced neuronal injury and that nimodipine and Co²⁺, by attenuating such influx, are able to attenuate Aβ neurotoxicity.     

Evaluation of biochemical methods for the non‐destructive identification of sex in upstream migrating salmon and sea trout

Female‐specific markers of reproductive activity [plasma 17β‐oestradiol (E2), vitellogenin (VTG) and alkali‐labile phosphoprotein phosphorous (ALP)] were measured over 12 months in a captive population of brown trout Salmo trutta . During the early months of the reproductive season (February to May) and using the concentration of plasma E2 or plasma ALP as a marker for females the proportion of fish in which sex was misidentified was high (15–50%). The misidentification rate was considerably lower (1–8%) using plasma VTG. Preliminary evaluation of a commercial immunochromatographic VTG test system as a screen for the presence or absence of VTG in plasma from brown trout provided results that were consistent with those obtained from direct measurement of plasma VTG levels by enzyme‐linked immunosorbent assay (ELISA). These preliminary conclusions were verified by sampling upstream‐migrating anadromous brown trout, sea trout, and Atlantic salmon Salmo salar trapped over a 6 month period. Plasma E2 levels did not satisfactorily discriminate between male and female sea trout and Atlantic salmon. Plasma VTG levels in both species, however, were bimodally distributed and it was assumed that this divergence corresponded to male (plasma VTG levels <10 μg ml⁻¹) and female (plasma VTG levels >800 μg ml⁻¹) fishes. Plasma ALP provided a more accurate indication of sex in the wild Atlantic salmon and sea trout than was suggested by the pilot study on captive brown trout. The commercial immunochromatographic VTG test system provided results that were wholly consistent with the data obtained from the trapped fishes by direct measurement of plasma VTG.