Heterofucan from Sargassum filipendula induces apoptosis in HeLa cells

Fucan is a term used to denominate a family of sulfated polysaccharides rich in sulfated l-fucose. Heterofucan SF-1.5v was extracted from the brown seaweed Sargassum filipendula by proteolytic digestion followed by sequential acetone precipitation. This fucan showed antiproliferative activity on Hela cells and induced apoptosis. However, SF-1.5v was not able to activate caspases. Moreover, SF-1.5v induced glycogen synthase kinase (GSK) activation, but this protein is not involved in the heterofucan SF-1.5v induced apoptosis mechanism. In addition, ERK, p38, p53, pAKT and NFκB were not affected by the presence of SF-1.5v. We determined that SF-1.5v induces apoptosis in HeLa mainly by mitochondrial release of apoptosis-inducing factor (AIF) into cytosol. In addition, SF-1.5v decreases the expression of anti-apoptotic protein Bcl-2 and increased expression of apoptogenic protein Bax. These results are significant in that they provide a mechanistic framework for further exploring the use of SF-1.5v as a novel chemotherapeutics against human cervical cancer.     

The effects of vehicle and region of application on in vitro penetration of testosterone through canine skin

The effects of the vehicles phosphate-buffered saline (PBS), ethanol (EtOH; 50% in PBS w/w) and propylene glycol (PG; 50% in PBS w/w) and the region of administration on in vitro transdermal penetration of testosterone was investigated in the dog. Skin was harvested from the thorax, neck (dorsal part) and groin regions of greyhounds after euthanasia and stored at -20 degrees C until required. The skin was then de-frosted and placed into Franz-type diffusion cells which were maintained at approximately 32 degrees C by a water-bath. Saturated solutions of testosterone, containing trace amounts of radiolabelled (14C) testosterone, in each vehicle were applied to the outer (stratum corneum) surface of each skin sample and aliquots of receptor fluid were collected at 0, 2, 4, 8, 16, 20, 22 and 24h and analysed for testosterone by scintillation counting. The maximum flux (J(max)) of testosterone was significantly higher for all sites when dissolved in a vehicle containing 50% EtOH or 50% PG, compared to PBS. In contrast, higher residues of testosterone were found remaining within the skin when PBS was used as a vehicle. This study shows that variability in percutaneous penetration of testosterone could be expected with formulation design and site of application.     

Persistent hyperplastic tunica vasculosa lentis and persistent hyperplastic primary vitreous (PHTVL/PHPV) in two cats

Two domestic shorthair cats (6 and 9 months old) were presented for examination of ocular opacities. One cat had bilateral persistent pupillary membranes, unilaterally accompanied by persistent hyperplastic tunica vasculosa lentis, persistent hyperplastic primary vitreous (PHTVL/PHPV), and cataract. The second cat had bilateral PHTVL/PHPV with dense white plaques in the posterior lens capsule and subcapsular cortex.     

Surgical treatment of disk-associated wobbler syndrome by a distractable vertebral titanium cage in seven dogs

Objective: To evaluate a distractable titanium cage for the treatment of disk‐associated wobbler syndrome (DAWS). Study Design: Prospective study. Animals: Dogs (n=7) with DAWS. Methods: After total discectomy of C5–C6 and C6–C7, the median part of the vertebral body of C6 was removed with preservation of the lateral walls and dorsal cortex. The removed cancellous bone was collected. The implant was placed in the bony defect of C6. After placement, the titanium cage was distracted and affixed by 4 screws. Finally, the implant was filled and covered with cancellous bone. Dogs had follow‐up examinations at 1, 3, 6, 12, and 24 months. Six months after surgery, cervical radiographs and computed tomography (CT) were performed. Results: Although no intraoperative complications occurred, correct placement of the cage was technically challenging. Revision surgery was necessary in 2 dogs because of implant loosening and aggravation of vertebral tilting. All dogs improved after discharge from the hospital. In 1 dog, recurrence of clinical signs caused by articular facet proliferation at an adjacent intervertebral disk space occurred. Radiographs at 6 months demonstrated cage subsidence in 4 dogs. In all dogs, CT was suggestive for fusion of the bone graft with the vertebral body. Conclusions: Although results are promising, technical adaptations will be necessary to make this specific surgical technique, designed for humans, suitable for routine use in dogs.     

In vivo measurement of central corneal thickness in normal chicks (Gallus gallus domesticus) with the optical low-coherence reflectometer

Objective To determine the practicability and accuracy of central corneal thickness (CCT) measurements in living chicks utilizing a noncontact, high‐speed optical low‐coherence reflectometer (OLCR) mounted on a slit lamp. Animals studied Twelve male chicks (Gallus gallus domesticus). Procedures Measurements of CCT were obtained in triplicate in 24 eyes of twelve 1‐day‐old anaesthetized chicks using OLCR. Every single measurement taken by OLCR consisted of the average result of 20 scans obtained within seconds. Additionally, corneal thickness was determined histologically after immersion fixation in Karnovsky’s solution alone (20 eyes) or with a previous injection of the fixative into the anterior chamber before enucleation (4 eyes). Results Central corneal thickness measurements using OLCR in 1‐day‐old living chicks provide a rapid and feasible examination technique. Mean CCT measured with OLCR (189.7 ± 3.34 μm) was significantly lower than histological measurements (242.1 ± 47.27 μm) in eyes with fixation in Karnovsky’s solution (P = 0.0005). In eyes with additional injection of Karnovsky’s fixative into the anterior chamber, mean histologically determined CCT was 195.2 ± 8.25 μm vs. 191.9 ± 8.90 μm with OLCR. A trend for a lower variance was found compared to the eyes that had only been immersion fixed. Conclusion Optical low‐coherence reflectometry is an accurate examination technique to measure in vivo CCT in the eye of newborn chicks. The knowledge of the thickness of the chick cornea and the ability to obtain noninvasive, noncontact measurements of CCT in the living animal may be of interest for research and development of eye diseases in chick models.     

Replication of genome-wide association signals in UK samples reveals risk loci for type 2 diabetes

The molecular mechanisms involved in the development of type 2 diabetes are poorly understood. Starting from genome-wide genotype data for 1924 diabetic cases and 2938 population controls generated by the Wellcome Trust Case Control Consortium, we set out to detect replicated diabetes association signals through analysis of 3757 additional cases and 5346 controls and by integration of our findings with equivalent data from other international consortia. We detected diabetes susceptibility loci in and around the genes CDKAL1, CDKN2A/CDKN2B, and IGF2BP2 and confirmed the recently described associations at HHEX/IDE and SLC30A8. Our findings provide insight into the genetic architecture of type 2 diabetes, emphasizing the contribution of multiple variants of modest effect. The regions identified underscore the importance of pathways influencing pancreatic beta cell development and function in the etiology of type 2 diabetes.     

Shelf‐edge reefs as priority areas for conservation of reef fish diversity in the tropical Atlantic

• 1. Data from fishing surveys employing bottom long‐lines were analysed to characterize the diversity, assemblages and distribution patterns of demersal fish along the Brazilian outer shelf and upper slope, between latitudes 13°S and 22°S.
• 2. Non‐metric multi‐dimensional scaling (MDS) and cluster analysis indicate three distinct species assemblages separated primarily by depth (the 200 m isobath) and secondarily by latitude (19°S), suggesting a continual transition along the depth and latitudinal gradients in the study area. Species richness was negatively correlated with depth, but with no clear relationship with latitude.
• 3. Results suggest the existence of reef formations on the shelf‐edge zone (40–200 m) and slope down to 500 m depth off the eastern Brazilian coast. More than 75% of the catches recorded were reef fish species from the families Serranidae, Lutjanidae, Malacanthidae, Muraenidae, Sparidae, Balistidae, Carangidae, Haemulidae, Scorpaenidae and Priacanthidae.
• 4. The maximum depth of occurrence for 20 reef species was extended from limits previously recorded.
• 5. The findings reinforce the hypothesis of a faunal corridor for species associated with deep reef formations along the shelf‐edge zone (40–200 m), in the South American continental margin, connecting the south‐western Atlantic and the Caribbean provinces.
• 6. The shelf‐edge reefs support important multi‐species fisheries and harbour critical habitats for the life cycle of many reef fish species, including spawning aggregation sites that are extremely vulnerable to human pressures, such as intensive fishing, shipping and offshore oil and gas exploitation; all activities currently expanding off the Brazilian coast.
• 7. Results reveal the biological importance of deep shelf‐edge reefs as a critical ecological area. Despite their importance, shelf edge reefs are not currently included in any marine protected area network in the tropical south‐western Atlantic. There is now an urgent need to enhance knowledge, implement adequate management strategies and consider these deeper habitats as priority areas for conservation. Copyright © 2011 John Wiley & Sons, Ltd.