The effect of photodynamic therapy on squamous cell carcinoma in a murine model: evaluation of time between intralesional injection to laser irradiation

Successful treatment of naturally occurring periocular squamous cell carcinoma (SCC) in horses with photodynamic therapy (PDT) has been performed by injecting residual tumor with verteporfin and applying laser irradiation immediately following injection. This study used a murine model to evaluate the influence of time between intralesional injection of verteporfin to laser irradiation on tumor growth inhibition with PDT. Mice were randomized into six groups (n=10/group). Each tumor was injected with either 0.1mg/cm(3) of verteporfin (Tx) or 5% dextrose in water (C). Tx and C groups 1, 2, and 3 were irradiated at 1, 30, and 180min after injection. Wilcoxon-rank sum test (P< or =0.05) was performed to determine the relative change in tumor volume (RCTV) between groups. Statistical significance was demonstrated between treatment groups. Although verteporfin-PDT treated mice in Tx1 and Tx2 demonstrated a lower RCTV compared to C1 and C2 mice, the differences were not statistically significant.     

In vitro embryo production in llamas (Lama glama) from in vivo matured oocytes with raw semen processed with Androcoll-E using defined embryo culture media

The aim of this study was to carry out in vitro fertilization using spermatozoa selected with Androcoll‐E? and to evaluate the efficiency of the culture medium DMEM‐F12 for in vitro embryo development in the llama. Twelve adult females from 18 superstimulated (67%) were used as oocyte donors. They were superstimulated with 1500 IU of eCG and after 5 days, received a single dose of buserelin. Twenty hours post‐injection, follicular aspiration was conducted by flank laparotomy. Semen collections were performed under general anesthesia by electroejaculation of the male. The ejaculates were processed with a solution of collagenase (0.1%) and an Androcoll‐E? column was used to improve the sample. Sixty nine COCs were recovered from 79 aspirated follicles (87% recovery). Only expanded COCs were used (n = 67); they were randomly placed in groups of 1–5 in Fertil‐TALP and the sperm suspension (20 × 10⁶ live spermatozoa/ml) was added to each fertilization microdroplet. After 24 h, they were randomly placed in one of two culture media: SOF (n = 34) or DMEM‐F12 (n = 33) and incubated for 6 days in humidified atmosphere of 5% CO₂, 5% O₂ and 90% N₂ at 38°C. The blastocyst rate was 20% (7/34) in SOF medium (3 hatched, 2 expanded and 2 early blastocysts) and 15% (5/33) in DMEM medium (all expanded blastocysts). In conclusion, using Androcoll‐E? it is possible to select good quality spermatozoa from llama ejaculates for in vitro fertilization and to produce blastocysts in DMEM‐F12 medium. This is also the first time that hatched llama blastocysts have been produced after culture in a defined medium such as SOFaa.     

Efficacy and safety of suberoylanilide hydroxamic acid (Vorinostat) in the treatment of canine corneal fibrosis

Objective Study aims were to evaluate the safety and efficacy of the Food and Drug Administration‐approved drug Vorinostat [suberoylanilide hydroxamic acid (SAHA)] in the treatment of canine corneal fibrosis using an in vitro model. Methods Healthy donor canine corneas were collected and used to generate primary canine corneal fibroblasts (CCFs) by growing cultures in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts, used as a model for corneal fibrosis, were produced by growing CCF cultures in serum‐free medium containing transforming growth factor β1 (1 ng/mL). Trypan blue exclusion assays were used to determine the optimal SAHA dose for this in vitro model. Four hour after culturing with TGFβ1, CCF cultures were treated with 0.06% SAHA for 5 min (group 1) and for 24 h (group 2), representing single and multiple dose treatment regimes, respectively. Cultures were then further incubated in the presence of TGFβ1 (1 ng/μL) under serum‐free conditions until they reached 70% confluence. Trypan blue exclusion, immunocytochemistry, and TUNEL assays were used to evaluate the cytotoxicity of SAHA. Real‐time PCR, western blot analysis, and immunocytochemistry were used to determine the efficacy of SAHA to inhibit canine corneal myofibroblast formation. Results Topical SAHA application in both treatment groups successfully decreased α‐smooth muscle actin expression when compared to the TGFβ1 only treatment group (P < 0.05). Tested SAHA did not affect CCF phenotype or cellular viability and did not cause significant cell death. Conclusions Suberoylanilide hydroxamic acid safely and effectively inhibits TGFβ1‐induced CCFs transformation to myofibroblast in vitro.     

Pistia stratiotes L. and Eichhornia crassipes (Mart.) Solms.: emerging invasive alien hydrophytes in Campania and Sardinia (Italy)

This article provides general information on the distribution and presence of invasive macrophytes in Italy and describes and discusses two hydrophyte invasion case studies: Eichhornia crassipes in Sardinia and Pistia stratiotes in Campania. The two invasions took place in the same period, but mechanical removal intervention started only in Sardinia, even if costly and unsuccessful in the long term. Two main pathways are responsible for the presence of these two species in Italy, i.e. introduction as ornamentals and investigation and use for phytoremediation. The drafting of a national strategy on biological invasions is a priority for Italy and several specific action plans for species and habitats, as in the case of inland waters, are required. Furthermore, there is the need for regulations in the trade sector of invasive plant species and to evaluate the possibilities for biological control for established aquatic alien invaders.     

Using multitemporal satellite imagery to characterize forest wildlife habitat: The case of ruffed grouse

Like many similar forest species, ruffed grouse (Bonasa umbellus; hereafter grouse) populations in the central and southern Appalachians (CSA) are strongly affected by forest composition at the landscape scale. Because these populations are in decline, managers require accurate forest maps to understand how stand level characteristics affect the survival and reproductive potentials of individual birds to design management strategies that improve grouse abundance. However, traditional mapping techniques are often labor-intensive and cost-prohibitive. We used a normalized difference vegetation index (NDVI) from each of 8 Landsat images and the digital elevation model (DEM)-derived variables of elevation and aspect in discriminant analyses to classify 7 study areas to 3 overstory classes (evergreen, hardwoods, and oak) and distinguish evergreen and deciduous understories in the CSA, 2000–2002. Overall accuracy was 82.08%, varying from 83.59% for oak to 79.79% for hardwoods overstories. Periods with large phenological differences among classes, particularly early and late spring, were most useful for discriminating overstory vegetation types. Alternatively, winter NDVI in combination with elevation was critical for differentiating evergreen and deciduous understories. Multitemporal image sets used in concert with DEMs provided a cost-effective alternative to hyperspectral sensors for improving wildlife habitat classification accuracy with Landsat imagery. This allowed for enhanced understanding of grouse-habitat relationships and habitat affects on grouse populations that allowed for improved management. With the incorporation of simple adjustments for local forest plant species phenology into the model, it may be used to better classify wildlife habitat of similar species in areas with comparable forest communities and topography. Multitemporal images can also be used to differentiate grassland communities, monitor wetlands, and serve as baseline data for detecting changes in land use over longer temporal scales, making their use in forest wildlife habitat studies cost-justifiable.     

In vitro efficacy of lufenuron against filamentous fungi and blood concentrations after PO administration in horses

Lufenuron is a benzoylphenyl urea-derived insecticide that has been recently introduced as a novel treatment for fungal infections in horses. The purposes of this study were to determine (1) the in vitro efficacy of lufenuron against Aspergillus and Fusarium spp. and (2) the ability of lufenuron to reach efficacious blood concentrations after PO administration in horses. Fungal colonies isolated from diseased equine corneas were tested against lufenuron solutions up to 700 microg/mL. Twenty-one adult horses received 1 of 3 PO lufenuron treatment regimens: 5 mg/kg body weight (BW) q24h for 3 days, 20 mg/kg BW q24h for 3 days, or 60 mg/ kg BW q24h for 1 day. Blood samples were collected up to 96 hours after drug administration and analyzed by high-performance liquid chromatography. Statistical analyses of lufenuron blood concentrations were performed by analysis of variance and Fischer's Least Significant Difference test, with statistical significance set at P < .05. Lufenuron showed no effect on the in vitro growth of Aspergillus or Fusarium spp. Lufenuron was detected in the blood of all but 1 horse and showed no adverse effects. The maximum blood lufenuron concentration (83.5 +/- 58.7 microg/L) was lower than the concentrations proven to be ineffective in vitro in this study. Further therapeutic use of lufenuron as an antifungal agent in horses should be based on proven efficacy against specific strains of clinically relevant fungi with pharmacokinetic data demonstrating sufficient lufenuron concentrations in target tissues.     

Retinal and optic nerve degeneration in cattle after accidental acrylamide intoxication

A herd of cattle was accidentally exposed to monomeric acrylamide and N-methylolacrylamide. Seven cattle underwent repeated examinations during 6–8 months after exposure. Abnormal pupillary light reflexes (PLR) were observed in one cow. Ophthalmoscopic examination showed progressive retinal degeneration and degenerative changes in the optic nerve heads in this cow. Light and electron microscopic examination revealed pathological changes in the retinae and optic nerves consistent with chronic stages of acrylamide toxicity. Ophthalmic examination and light microscopy did not reveal abnormalities in the retinae or optic nerves in either the rest of the cattle from the herd or in offspring of exposed cattle.     

Morphology,morphometry and chromatin distribution in llama sperm nuclei

The objectives of this study were as follows: (i) to describe and evaluate the frequencies of different morphologies of llama sperm nuclei, (ii) to determine morphometric values of nuclear parameters, (iii) to describe and estimate the frequencies of different classes of chromatin distribution and (iv) to measure haploid DNA content and analyse its nuclear distribution. The study was performed using ejaculates collected from seven males, and sperm nuclei were stained with the Feulgen reaction. Normal morphology ranged from 78.36% to 93.92%, and abnormalities included short, small, large, pyriform, narrow, micro and round nuclei. Important differences in nuclei considered normal were found between some males. The following average values were obtained for each sperm nuclear morphometric parameter analysed: area 11.64 μm², perimeter 13.16 μm, length 5.12 μm, width 2.81 μm, ellipticity 1.85 and form 0.83. Differences between males were significant for all the parameters (p < .01). Light microscope observations and cytophotometric determinations allowed discriminating between three classes of chromatin distribution: homogeneous, diffuse and showing a clear band. Significant differences between males were found for the frequencies of the three classes (p < .01). Cluster analysis methods were used to estimate the resemblance between males according to the characteristics of their sperm nuclei. A great intermale variability was found for morphological, morphometric and chromatin distribution data. These parameters would have low dependence between them.