Monoclonal antibodies identifying chick gonadal cells

1. The aim of the present study was to raise monoclonal antibodies (MAbs) which discriminate gonadal cell types in chickens.

2. A panel of 180 MAbs was generated, from which 14 were chosen for their specificity.

3. These MAbs were characterised immunohistologically on frozen sections of embryonic, newly‐hatched and adult, male and female, left gonads.

4. Three of these MAbs are described: AGC5, AGC7, and AGC13 which recognise respectively germinal epithelial cells, supporting cells and germ cells.

5. These MAbs may be useful in following the developmental pathways in the chicken gonad and analysing the interactive role(s) taking place between gonadal stromal cells and primordial germ cells.     

Effects of formaldehyde fixation on equine platelets using flow cytometric methods to evaluate markers of platelet activation

OBJECTIVE: To investigate the effects of formaldehyde fixation on equine platelets using flow cytometric methods to evaluate markers of platelet activation. SAMPLE POPULATION: Blood samples from 6 Thoroughbreds. PROCEDURE: The degree of fluorescence associated with binding of fluorescein isothiocyanate (FITC)-conjugated anti-human fibrinogen antibody and FITC-annexin V in unactivated and adenosine diphosphate (ADP)-, platelet activating factor (PAF)-, and A23187-activated platelet samples in unfixed and 0.5, 1.0, and 2.0% formaldehyde-fixed samples was assessed by use of flow cytometry. RESULTS: In samples incubated with FITC-anti-human fibrinogen antibody prior to fixation, addition of 2.0% formaldehyde resulted in a 30% increase in total fluorescence in ADP- and PAF-activated samples and a 60% increase in A23187-activated samples. Fixation for 24 hours prior to addition of antibody resulted in reduced fluorescence of samples containing antihuman fibrinogen antibody for all 3 concentrations of formaldehyde in PAF-activated samples. The addition of all 3 concentrations of formaldehyde after incubation with FITC-annexin V resulted in significant increases in fluorescence in unactivated and activated platelet samples. As length of fixation time increased, there was a gradual increase in fluorescence that was significant at 24 hours. CONCLUSIONS: Because fixation with 2.0% formaldehyde results in significant changes in fluorescence in activated platelet samples containing anti-fibrinogen antibody, lower concentrations of formaldehyde should be used to fix equine platelet samples. Formaldehyde-fixed platelet samples should be analyzed within 12 hours of fixation to avoid artifactual increases in fluorescence. Fixation of samples containing FITC-annexin V should be avoided because of significant increases in fluorescence that may interfere with interpretation of results.     

Effects of intravenous administration of formaldehyde on platelet and coagulation variables in healthy horses

OBJECTIVES: To assess safety and determine effects of IV administration of formaldehyde on hemostatic variables in healthy horses. ANIMALS: 7 healthy adult horses. PROCEDURE: Clinical signs and results of CBC, serum biochemical analyses, and coagulation testing including template bleeding time (TBT) and activated clotting time (ACT) were compared in horses given a dose of 0.37% formaldehyde or lactated Ringer's solution (LRS), IV, in a 2-way crossover design. In a subsequent experiment, horses received an infusion of 0.74% formaldehyde or LRS. In another experiment, horses were treated with aspirin to impair platelet responses prior to infusion of formaldehyde or LRS. RESULTS: Significant differences were not detected in any variable measured between horses when given formaldehyde or any other treatment. Infusion of higher doses of formaldehyde resulted in adverse effects including muscle fasciculations, tachycardia, tachypnea, serous ocular and nasal discharge, agitation, and restlessness. CONCLUSIONS AND CLINICAL RELEVANCE: Intravenous infusion of formaldehyde at doses that do not induce adverse reactions did not have a detectable effect on measured hemostatic variables in healthy horses.     

Growth of young cuttlefish, Sepia officinalis (Linnaeus 1758) at the upper end of the biological distribution temperature range

Individual growth rates, feeding rates (%BWd^?1) and food conversions for cuttlefish (S. officinalis) hatchlings and juveniles were determined during this study. A flow‐through system was used. Water temperature reached 30 °C during the hottest part of the day, gradually decreasing to 25 °C during the night; salinity varied between 37 ± 3 ppt and lights were kept on for 14 h day^?1. Hatchlings were placed in separate compartments with a water volume of 1.2 L. Juvenile cuttlefish (from 0.5 to 25 g) were placed in bigger baskets, with a water volume of 5.2 L. Water flow was 120 L h^?1. The biggest cuttlefish used in these experiments (> 25 g) were gathered in groups of five and placed in circular tanks (water volume of 250–300 L). Thus, results obtained in this case are means and not individual data. During the first 10, 20, 30 and 40 days, mean growth rates (of all individuals sampled by age group) decreased consistently (11.8 ± 4.1, 9.8 ± 1.8, 8.1 ± 2.2 and 7.3 ± 0.7%BW^?1 respectively); in similar fashion, mean feeding rates decreased with age group (33.7 ± 13.5, 22.0 ± 7.9, 17.3 ± 3.9 and 16.7%BWd^?1 respectively). Mean food conversions varied between 3.6 and 2.5 between the age groups. When grouping results by weight class, similar patterns occur, as growth and feeding rates decrease consistently as cuttlefish grow bigger. Highest mean growth and feeding rates are obtained by hatchlings (< 0.1 g) with 12.4 ± 4.5 and 35.3 ± 15.1%BWd^?1, respectively, while the lowest growth and feeding rates were recorded for the largest animals, between 15 and 25 g (3.4 ± 1.1 and 10.8 ± 4.1%BWd^?1 respectively). For these weight classes, mean food conversions varied between 2.7 ± 0.9 and 3.8 ± 2.8.     

Dramatic decline in bat species richness in Singapore, with implications for Southeast Asia

Literature searches and recent surveys of the bat fauna of Singapore indicate that of the 24 species of Microchiroptera and six species of Megachiroptera documented for this small equatorial island just 15 and 5, respectively, are still present. These recorded declines in chiropteran species richness almost certainly understate the true losses as extensive land transformation/habitat loss (>95%) and biota loss occurred early in Singapore’s colonial history before comprehensive surveys of bats were made. In an effort to reconstruct the pre-settlement bat fauna, we inferred an upper bound of pre-settlement species richness using a checklist from a well-known bat assemblage in neighbouring Peninsular Malaysia, and a lower bound based on species common to Peninsular Malaysia, Borneo and Sumatra. The Singapore records were compared with these two species list predictions. Based on this analysis, we infer that between 60 and 72 species would have inhabited Singapore before 1819. We also estimate that between 33% (based on the confirmed inventory) and 72% of the species (based on the upper-bound estimate of species richness) are now locally extinct. For Microchiroptera the data suggest that the documented local extinction rate of 38% may project to between 69% and 75%. Forest-dependant bats are particularly affected and comprise a much lower proportion of the bat fauna than is seen in intact forest in Peninsular Malaysia. All hipposiderids and 40% of the documented rhinolophid taxa have been lost and almost half (6) of the surviving microchiropteran species are locally endangered. Projected local extinction rates for Megachiroptera raise the documented value (17%) to about 60%, with most of the survivors being widespread species known to forage in cultivated or secondary forest habitats or to commute long distances between fragmented resources. The dramatic decline in Singapore bat species richness and a concomitant change in chiropteran guild and trophic structure (Microchiroptera vs. Megachiroptera) reflect patterns of diversity change seen elsewhere in the region in response to loss of forest habitat. In Singapore, the decline in diversity (species richness and abundances) for both mega- and microbats may also relate to urbanisation and deliberate or accidental destruction of bats and their roost sites in a land that has one of the highest human population densities on the planet. Although these losses (actual and inferred) represent a microcosm of mainly local extinctions, a wider geographical extrapolation over the 21st century indicates that heavy deforestation in progress in Southeast Asia might be expected to lead to extinction of many bat taxa, with upper-bound estimates of regional species losses exceeding 40% and global extirpation anticipated for at least 23% of Southeast Asia’s bat fauna by 2100.     

Avian whole‐brain spheroid cultures: applications in pesticide toxicity

The adult hen is the species of choice when assessing the potential of organophosphate compounds (OPs) to evoke OP‐induced delayed neuropathy (OPIDN). OPIDN has been shown to depend on the irreversible covalent binding of certain OPs to the enzyme neuropathy target esterase (NTE). Levels of this enzyme in human brain are similar to those in avian species. We have previously shown that the hen embryo whole brain spheroids are histotypically similar to adult brain and express measurable levels of acetylcholine esterase (AChE) and NTE, the key enzymes in mechanisms of OP toxicity. Single‐cell suspensions were prepared from meninges‐free 7 day in ovo hen brains. Cells were grown in serum‐free media and incubated on an orbital shaker at 37 °C in 5% carbon dioxide humidified air. Spheroids can be maintained long‐term in culture and allow the effects of repeated dose and recovery studies to be conducted. Hen embryo brain spheroids were exposed to varying concentrations of tri‐o‐cresyl phosphate, leptophos and its metabolite leptophos oxon at day 14 in vitro and the effects on the activity of AChE and NTE measured. Whilst current legislation regarding toxicity testing of OPs requires the use of animals, our in vitro model provides a potential pre‐screen for novel chemical entities and commercial OP mixture variants. Such an approach should refine and reduce the number of animals used in development of new materials. © 2000 Society of Chemical Industry     

The Chemistry of Intercepted Clouds in Northern Arizona during the North American Monsoon Season

Cloudwater samples have been collected for the first time at a high-elevation site in the US interior Southwest. Cloud samples were collected at the summit of Mt. Elden near Flagstaff, Arizona. The samples were analyzed for pH, ionic composition, trace metals, organic carbon content, and volatile organic compounds. All of the samples showed high pH values (5.12–6.66), which appear to be the result of soil/crustal acid-neutralizing components. Ammonium and nitrate were the dominant ionic species. Organic carbon concentrations ranged from 3 to 18 mg/l. Volatile aromatic compounds (toluene, ethylbenzene, and xylenes) were detected, although they did not contribute significantly to the dissolved organic matter (<1% of dissolved organic carbon). Still, their aqueous-phase concentrations were substantially higher than equilibrium partitioning from the gas phase would suggest. Metal concentrations were high when compared to other cloud studies in remote areas. Overall, with the exception of pH, the cloud chemistry showed marked inter-event variability. The source of the variability was investigated using NOAA HYSPLIT dispersion calculations. Like the cloud composition, the air mass back trajectories differed widely from event to event, and consistently, air masses that passed over highly urbanized areas had higher trace metal, organic, and ion concentrations than more pristine air masses.     

Review of the sufficiency of research on the swelling pressure of wood

Summary Terms used in relation to swelling pressure are first defined. The present status of knowledge of the swelling pressure of wood under various conditions is then briefly discussed.Methods are outlined for the determination of swelling pressure with restraint in one direction only and their limitations discussed. The method developed by Tarkow and Turner for the determination of swelling pressure when restraint is present in two directions is also referred to and outlined.Conclusions are then drawn from the present status of knowledge of swelling pressure in wood and gaps in this knowledge are indicated.     

Identification of two compounds correlated with lack of rooting capacity of chestnut cuttings

Cuttings from mature chestnut (Castanea spp.) trees are very difficult to root, whereas juvenile or etiolated cuttings root easily. The HPLC chromatograms from mature cuttings consistently showed two well-defined peaks that were absent from juvenile or etiolated cuttings. The compounds responsible for these two peaks were isolated and identified as derivatives of ellagic acid, 3,3',4-tri-O-methylellagic acid (compound 1) and 3,4,4'-tri-O-methylellagic acid (compound II). Compound I significantly inhibited rooting as measured by the bean test; a concentration of 3.0 x 10(-5) M caused a 32% inhibition in number of roots formed compared with controls grown in 10(-5) M IAA. When compound I was mixed with an equal quantity of IAA, the rooting activity of the auxin was reduced by 31.5%. An insufficient amount of compound II was available for the bean assay.