Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

Assessing seed desiccation responses of native trees in the Caribbean

New Forests - Native trees from the Caribbean were tested for seed desiccation responses, by adapting the “100-seed test” protocol. Ninety-seven seed lots of 91 species were collected...     

Synthesis and in-vitro antifungal activity of 6-trifluoromethyipyrazolo[3,4-6d]pyrimidin-4(5H)-thiones

Twelve 6-trifluoromethylpyrazolo[3,4-d]pyrimidin-4(5H)-thiones were prepared by the reaction between 4-thiocarbamoyl-5-aminopyrazoles and trifluoroacetic anhydride. They were tested in vitro for antifungal activity against a series of phytopathogenic fungi of different taxonomic classes. The EC50 and MIC values of four compounds were comparable or inferior to those of reference commercial fungicides in controlling Sclerotinia minor, Corticium solani and Phytium ultimum.     

Fungitoxicity of 5-aminoisoxazole-4-thiocyanate derivatives

3-Methyl-5-(acyl/alkyl)aminoisoxazole-4-thiocyanates were prepared by rhodanation of N-acyl/alkyl-3-methyl-5-isoxazolamines. These products were tested for antifungal activity against a series of phytopathogenic fungi of different taxonomic classes. Some of the compounds showed interesting in-vitro activity. The more active compound in the in-vitro test displayed a moderate preventive activity against Plasmopara viticola, Alternaria solani and Pyricularia recondita.     

Benthic microbial indicators of fish farm impact in a coastal area of the Tyrrhenian Sea

We studied the impact of organic loads due to the biodeposition of a fish farm in a non-impacted coastal area of the Tyrrhenian Sea (Western Mediterranean). Sediment chemistry and benthic microbial community were investigated from July 1997 to February 1998 on monthly basis at two stations: one was located under the fish farm, while the second was about 1 km away, and served as a reference site. The presence of the cage induced rapid changes in the benthic conditions: the sediments were rapidly (after 6 weeks) reduced. A significant accumulation of biopolymeric carbon was observed beneath the cage both 2 weeks after the initial cage deployment, on non-impacted sediments, and 5–7 months after, and appeared to be related to the fish farm production cycle. The density of microbial communities beneath the fish farm increased only during the first month of farming activity (July) and partially 7 months after cage disposal (i.e. in January), when there was a significant sediment organic enrichment. Additional evidence of rapid impact of the fish farm on the benthic bacterial communities is provided by the increase in the numbers of autofluorescent microbial cells. The photosynthetic eukaryotic cells displayed a highly reduced contribution to total autofluorescent microbial density, indicating that their decrease is related with biopolymeric carbon accumulation, and probably was due also to “shadow effect” induced by suspended material, coming from the fish farm, on the environment below. We propose here, to apply the ratio of culturable heterotrophic bacteria to microbial direct counts (CFU/MDC) to detect fish farm impact. In cage sediments, organic enrichment and the consequent modification of the characteristics of the benthic environment, determined an increase in aerobic heterotrophic bacteria and vibrio density indicating that they are efficient colonizers of organic-rich sediments. Densities of Escherichia coli and Enterococci were not significantly higher than in the reference site, and are likely of terrestrial origin. Thus they cannot be used as specific indicators of fish farm impact.     

Use of internal splinting to realign a noncompliant sternum in a cat with pectus excavatum

CASE DESCRIPTION: A 5.5-month-old female domestic longhair cat was examined because of dorsal deviation of the caudal aspect of the sternum and progressively worsening dyspnea during play activities. CLINICAL FINDINGS: A diagnosis of pectus excavatum was made clinically and confirmed radiographically. The cardiac silhouette was shifted into the right hemithorax. The vertebral index (VI) and frontosagittal index (FSI), which are radiographic indices used to grade the extent of the deformity, were 5.2 and 3.0, respectively. Minimum thoracic height was 13 mm at this time. TREATMENT AND OUTCOME: The sternal malposition was corrected during surgery; for stabilization, an internal splint was provided by use of a plate that was applied to the ventral side of the sternum. Radiographic indices improved until day 85; at that time, an increase in the distance between plate and sternum was detected, and plate removal was advised. The radiographic indices improved again after plate removal. At 310 days after surgery, VI was 9.6, FSI was 1.6, and minimum thoracic height was 34 mm. No dyspnea was evident during physical examination. CLINICAL RELEVANCE: Results of treatment suggest that this technique may be useful as an alternative surgical option for cats with pectus excavatum that have a noncompliant sternum.     

Nutrition and feeding of the geriatric horse.

Little is known regarding nutrient requirements and feeding of geriatric horses, and more effort should be placed on this area of equine nutrition research. That which is known suggests that some geriatric horses may not have different requirements than other mature horses, whereas others affected by disease or poor dentition may have special nutritional needs. In general, rations for geriatric horses should be based on high-quality roughage supplemented with complementary minerals and vitamins. The need for additional energy aside from that provided by the forage can be supplied by adding energy concentrates, such as cereal grains or fat, to the ration. Processing techniques involving heat, such as pelleting and extruding, are advised when cereal grains are included in the ration so as to improve starch digestibility in the small intestine and avoid starch overload in the hindgut and it subsequent problem (ie, colic, laminitis). In addition, the environment in which geriatric horses are fed should be one that promotes ease of ration consumption and eliminates factors thar impair feed consumption, such as competition from other horses and the need to travel relatively long distances (eg, grazing marginal pastures). Finally, strict attention should be paid to the body condition of geriatric horses so as to evaluate adequacy of the ration and the general health of the horse.     

Effect of Restricted Pasture Access on Pasture Dry Matter Intake Rate,Dietary Energy Intake,and Fecal pH in Horses

Eight mature horses were used in a 4 × 4 Latin square design to determine the effect of restricted pasture access on dry matter intake rate (DMIR), energy intake, and fecal pH. Horses were randomly assigned to one of four groups (HGRPs), each containing two horses. HGRPs were randomly assigned to one of four treatments consisting of 3, 6, 9, or 24 hours of pasture access, for a period of 7 days. Treatments were switched every 7 days. The 3- and 6-hour groups were fed free choice mature grass hay while not grazing. Daily pasture dry matter intake (DMI) for each HGRP was estimated by calculating the difference between initial herbage mass of the grazing cell before grazing and residual herbage mass after 7 days of grazing. Total dry matter intake was the sum of pasture and hay DMI for the 3- and 6-hour treatments. Fecal pH was measured on day 7 of each period. Response variables were analyzed using analysis of variance for Latin square design. Restricting pasture access decreased mean pasture DMI (P = .02), pasture dietary energy (DE) (P = .02), and fecal pH (P < .001), but increased DMIR (P = .02). Mean total dry matter intake was not different (P = .16) among treatments. In conclusion, restricting pasture access accelerates pasture DMIR and decreases fecal pH, and although restricting pasture access decreases DE intake from pasture, it did not result in a decrease in total DE intake for horses having ad libitum access to hay.