A new species of Acanthocotyle Monticelli, 1888 (Platyhelminthes: Monogenea: Acanthocotylidae) from the ventral skin of the banded stingaree, Urolophus cruciatus (Lacépède, 1804), from Tasmania, Australia.

Acanthocotyle urolophi sp. nov. is described from the skin of the banded stingaree, Urolophus cruciatus (Lacépède, 1804). This is the first acanthocotylid to be described from Australian waters. Acanthocotyle urolophi sp. nov. is distinguished from other species of Acanthocotyle by a combination of the number of vitelline follicles 38 (33-46) and the number of rows 35 (32-37) of sclerites on the pseudohaptor. In addition, Acanthocotyle urolophi sp. nov. has no germarial appendix and no uterine "arm". A uterine receptaculum seminis was not identified in whole mounts. There is no penis papilla and no penis sclerite associated with the male reproductive opening. A brief description of the larva is provided. The diagnosis of the Acanthocotylidae Price, 1936 is amended and we review the Allacanthocotylinae Yamaguti, 1963, Lophocotylinae Yamaguti, 1963 and Pseudacanthocotylinae Yamaguti, 1963. We deem that these subfamilies are invalid and that the family now comprises only the subfamily Acanthocotylinae and the genus Acanthocotyle. The validity of species previously assigned to the Acanthocotylidae (sensu Yamaguti, 1963) is discussed and a key to what we consider to be the valid species in the family is also provided.

A new species of Entobdella Blainville in Lamarck, 1818 (Monogenea: Capsalidae: Entobdellinae) from the Greenland halibut, Reinhardtius hippoglossoides.

A previously undescribed species of Entobdella is reported from the skin of the Greenland halibut, Reinhardtius hippoglossoides (Pleuronectiformes, Pleuronectidae). Entobdella whittingtoni sp. nov. differs from other species of Entobdella, including skin parasites of the related pleuronectids Hippoglossus hippoglossus (Atlantic halibut) and H. stenolepis (Pacific halibut), in lacking papillae on the ventral surface of the haptor. Other characteristics of E. whittingtoni are as follows: the absence of vitelline follicles lateral to the pharynx thereby exposing gut caeca in this region of whole mounts; the presence of a circular feature of unknown function, resembling a rosette in sections, attached to the wall of the internal male accessory reservoir; the lack of eyes. Papillae are also absent from the ventral surface of the haptor of the gill-parasitic entobdelline Branchobdella pugetensis, a gill parasite of the pleuronectid Atheresthes stomias. This raises the question as to whether this gill parasite has evolved from a skin-parasitic ancestor similar to E. whittingtoni. An answer to this question requires a more detailed study of the male reproductive apparatus of B. pugetensis and the use of molecular techniques to explore the relationship between B. pugetensis and E. whittingtoni.

Effect of total lens epithelial cell destruction on intraocular lens fixation in the human capsular bag.

PURPOSE: To evaluate the effect of complete destruction of lens epithelial cells (LECs) in the capsular bag on intraocular lens (IOL) stability. SETTING: School of Biological Sciences, University of East Anglia, Norwich, United Kingdom. DESIGN: Comparative evaluation. METHODS: An in vitro organ culture model using the bag-zonule-ciliary body complex isolated from fellow human donor eyes was prepared. A capsulorhexis and fiber extraction were performed, and an Acrysof IOL was implanted. Preparations were secured by pinning the ciliary body to a silicone ring and maintaining it in 6 mL Eagle minimum essential medium supplemented with 5% v/v fetal calf serum and 10 ng/mL transforming growth factor-ß2 for 3 weeks or more. One bag of each pair was treated with 1 µM thapsigargin to destroy all LECs. Observations of LEC growth were captured by phase-contrast microscopy, IOL stability by video microscopy, and endpoint analysis through scanning electron microscopy and immunocytochemistry. RESULTS: The LECs in control capsular bags migrated centrally, closing the bag and fixating the IOL between the anterior and posterior capsules, as seen clinically. These events were not observed in the thapsigargin-treated group. After a period of controlled orbital movement, the IOL in the control group stabilized quicker than in the treated bags. There was no IOL rotation in the bag; however, the IOLs in the treated group rocked with axial movement. CONCLUSIONS: The LECs appeared to aid stabilization of current IOL designs in the capsular bag. The results have clinical implications for IOL design and for strategies to prevent posterior capsule opacification. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Chemically-coupled-peptide-promoted virus nanoparticle templated mineralization.

The external surface of the plant virus Cowpea mosaic virus (CPMV) can be chemically modified with peptides that direct specific mineralization processes. Subsequent mineralization of the peptide-CPMV conjugates produces monodisperse nanoparticles of ca. 32 nm diameter coated with, for example, cobalt-platinum, iron-platinum or zinc sulfide, which cannot be readily prepared by other methods. This route is particularly attractive as it avoids the need to genetically engineer the protein surface of the virus to provide chimaeras for templated-mineralization. The synthetic procedure is environmentally friendly, as it proceeds at ambient temperature and pressure, in aqueous solvent. Further, the methodology is demonstrated to be generally applicable by the mineralization of a peptide-modified multiwalled carbon nanotube.

A new method to obtain epithelial and stromal explants from human Corneo-Scleral Discs for the routine culture of corneal epithelial and fibroblast cells.

Acquisition of human corneal cells for culture is hindered not only by the scarcity of donor tissues but also by some of the standard enzymatic and mechanical isolation techniques. Good yields have been reported from full-thickness explant and sclero-limbal pieces. However, due to their greater proliferative capacity, fibroblasts will encroach and subsequently overwhelm epithelial cultures whichever technique is used. The novel approach presented here is to minimise this by removal of the whole stroma from the epithelial layers at the outset. This is achieved by selective sectioning with the Webb mini-microtome developed in the Norwich Eye Research Laboratory. The microtome can be sterilised by alcohol spraying or autoclaving and is small enough to use in the culture hood. A selective cut in the region of the Bowman's membrane results in the isolation of the epithelium from the stroma and thus exposed, the basal epithelial layers are released from contact inhibition to allow growth. The stroma is further cut to produce multiple sections for the culture of fibroblasts. Both pure epithelial and stromal fibroblast cultures have been successfully generated in serum-enriched medium as well as defined serum-free media with growth supplements, from the corneo-scleral discs of donors of all ages.

Cadmium-specific formation of metal sulfide 'Q-particles' by Klebsiella pneumoniae.

Klebsiella pneumoniae overcomes cadmium toxicity through the 'biotrans-formation' of cadmium ions into photoactive, nanometre-sized CdS particles deposited on the cell surface. The kinetics of particle formation during batch culture growth was monitored by electron microscopy (EM), energy-dispersive X-ray analysis and electronic absorption spectroscopy (EAS). During the deceleration phase of bacterial growth, the presence of CdS particles on the outer cell wall of K. pneumoniae (> or = 5 nm in diameter) was detected by EM. The size of these electron-dense particles continued to increase throughout the stationary phase of growth, with some of the particles reaching a diameter > 200 nm. The formation of the extracellular CdS particles contributed to around 3-4% of the total cell biomass. EAS undertaken on these extracellular 'bio-CdS' particles suggested that the large 'superparticles' observed by EM, e.g. 200 nm, were aggregates of smaller particles termed 'Q-particles', approximately 4 nm in diameter. Metal sulfide particles were not formed in batch cultures of K. pneumoniae grown in the presence of lead, zinc, mercury, copper or silver ions. Growth in the presence of lead ions resulted in the formation of extracellular electron-dense particles containing lead but not sulfide or phosphate. Intracellular phosphorus-containing electron-opaque particles were formed during growth in the presence of copper and mercury. Intracellular electron-dense particles were formed in the presence of zinc ions but these did not contain phosphorus. From these results it was thought that metal sulfide formation in K. pneumoniae showed some cadmium-specificity. When cadmium and zinc ions were both added to the growth medium, metal sulfide particles were formed that were predominantly composed of cadmium, e.g. 48.6% cadmium and 0.04% zinc. Similarly, when cadmium and lead ions were both present during growth only CdS particles formed. In both cases analysis of the cells by EAS confirmed the presence of CdS only. These observations suggest that the mechanism of CdS formation is unlikely to occur simply through a cadmium-induced release of hydrogen sulfide by the cells into the external environment. If hydrogen sulfide production was the mechanism of sulfide formation then metal sulfide particles containing lead and zinc ions in addition to cadmium ions should have been produced.