Surgical Retrieval,Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications

Injury to the ACL is a commonly encountered problem in active individuals. Even partial tears of this intra-articular knee ligament lead to biomechanical deficiencies that impair function and stability. Current options for the treatment of partial ACL tears range from nonoperative, conservative management to multiple surgical options, such as: thermal modification, single-bundle repair, complete reconstruction, and reconstruction of the damaged portion of the native ligament. Few studies, if any, have demonstrated any single method for management to be consistently superior, and in many cases patients continue to demonstrate persistent instability and other comorbidities.The goal of this study is to identify a potential cell source for utilization in the development of a tissue engineered patch that could be implemented in the repair of a partially torn ACL. A novel protocol was developed for the expansion of cells derived from patients undergoing ACL reconstruction. To isolate the cells, minced hACL tissue obtained during ACL reconstruction was digested in a Collagenase solution. Expansion was performed using DMEM/F12 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S). The cells were then stored at -80 ºC or in liquid nitrogen in a freezing medium consisting of DMSO, FBS and the expansion medium. After thawing, the hACL derived cells were then seeded onto a tissue engineered scaffold, PLAGA (Poly lactic-co-glycolic acid) and control Tissue culture polystyrene (TCPS). After 7 days, SEM was performed to compare cellular adhesion to the PLAGA versus the control TCPS. Cellular morphology was evaluated using immunofluorescence staining. SEM (Scanning Electron Microscope) micrographs demonstrated that cells grew and adhered on both PLAGA and TCPS surfaces and were confluent over the entire surfaces by day 7. Immunofluorescence staining showed normal, non-stressed morphological patterns on both surfaces. This technique is promising for applications in ACL regeneration and reconstruction.     

Seasonal movements of British Greenfinches Carduelis chloris

Ringing recoveries are used to investigate the seasonal movements of British Greenfinches. Significantly more females than males make such movements. Outward movement begins by early November and continues until at least late January. The return movement can last from January to May and is particularly rapid in March and early April. The directions of the outward movements are widely dispersed but show trends towards the south and, except in western areas, towards the west. In Britain south of latitude 54°N the proportion of birds leaving their breeding grounds in winter increases from west to east, being negligible in the south-west and largest in East Anglia, but relatively small near the south coast. Distances moved show a similar dependence. The numbers making seasonal movements in a given year are correlated with the breeding population, particularly in south-east England.     

Effects of a synthetic juvenile hormone and some analogues on Ephestia spp. (Lepidoptera: Phycitidae)

Depending on their stage of development, treatment of mature larvae of Ephestia kühniella with a synthetic juvenile hormone resulted in the production of super larvae (which invariably prolonged larval life) and larval-pupal intermediates. When migrating last-instar larvae were treated with the juvenile hormone analogues (JHA) ethyl-3,7, n-trimethyldodeca-2,4-dienoate (ZR512) and isopropyl ii-methoxy-3,7, ii-trimethyldodeca-2,4-dienoate (ZR515), larval-pupal intermediates and pupal mortality were induced. However, when applied topically, ZR515 appeared more effective than ZR512. Both analogues prevented adult emergence when topically applied to the migrating larvae at doses between 28–52 ng. One-day-old pupae were most susceptible while older individuals became less sensitive with age. When larvae pupated in corrugated cardboard rolls were treated with ZR512 those of both E. cautella and E. kiihniella failed to emerge. At an estimated dose of 179 ng cm^-2, ZR515 prevented 77^-6%E. cautella and 100%E. kühniella larvae from emerging as adults. The control of Ephestia by JHA treatment of the pupation sites is discussed.     

Apolipoprotein E genotyping using the polymerase chain reaction and allele-specific oligonucleotide primers

A method for apolipoprotein (apo) E genotyping was developed using the polymerase chain reaction (PCR) with allele-specific oligonucleotide primers (ASP). Synthetic oligonucleotides with base-pair mismatches at the 3' terminus were used as primers to amplify the apoE gene in subjects previously phenotyped using isoelectric focusing (IEF). Complementary primer-allele combinations were specifically amplified by PCR, together with a control pair of primers specific to the human prothrombin gene. Identification of genotype by PCR using ASP was consistent with the phenotypes that were determined by IEF for 14 healthy normolipidemic subjects. These results were achieved using DNA isolated from buccal epithelial cells obtained from a mouthwash or DNA extracted from leukocytes. Genotype identification required analysis of the PCR products on an ethidium-stained agarose gel, yielding results 3 h after DNA extraction. In comparison with other current methods, PCR using ASP is suggested as a rapid and simple noninvasive technique for determining population apoE allelic distribution.     

THE FOOD AND REPRODUCTIVE CYCLES OF THE CENTIPEDES LITHOBIUS VARIEGATUS AND LITHOBIUS FORFICATUS IN A YORKSHIRE WOODLAND

Lithobius variegatus and L. forficatus occupy similar ecological niches and frequently occur together although there are differences in their distribution in the British Isles. An investigation of their food and reproductive cycles, being a preliminary step towards a further understanding of their ecological relationships, is described.
Lithobiomorph centipedes have hitherto been regarded as wholly carnivorous, but it is shown that these species feed on litter as well as small litter animals, the presence of litter in the guts of these species not being connected with the presence of animal remains. Whereas L. forficatus takes litter throughout the year, L. variegatus does so mainly in the winter.
Both species appear to lay eggs for a considerable part of the year, though there appears to be only a short period of sperm transfer in spring. It is suggested that the long oviposition period may prevent the loss of the entire brood in a dry summer. The succession of epimorph stadia in L. variegatus is described.
L. forficatus was more common than L. variegatus in the drior part of the woods studied.