Die Bek?mpfung der FichtengallenlausAdelges laricis Vall. (Anlage und Auswertung eines Versuches)

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Protura (Beintastler)

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Effects of growth factors (EGF,PDGF-BB and TGF-beta 1) on cultured equine epithelial cells and keratocytes: implications for wound healing

Objective The physiologic mechanisms involving growth factors, including PDGF‐BB, EGF, and TGF‐β1, as potent mediators of fibroblasts and epithelial cells in corneal wound healing remain unknown. The goal of this study was to determine culture methods for equine epithelial cells and keratocytes and to investigate how exogenous growth factors influence proliferation of both cell types. Procedures Cell cultures were established from healthy corneas harvested from horses immediately following euthanasia and maintained using standard tissue culture protocols. To determine the effects of PDGF‐BB, EGF, TGF‐β1, keratocytes (1 × 10⁵/well) and epithelial cells (2 × 10⁵/well) were each cultured in 12 well plates and exposed separately to the growth factors. The cells were exposed to concentrations of EGF between 0 and 50 ng/mL; PDGF‐BB between 0 and 75 ng/mL; and TGF‐β1 between 0 and 10 ng/mL. Cell proliferation was measured using ³H‐thymidine assay and differences in growth determined using anova and Tukey's HSD test (P < 0.05). Results Epithelial cell and keratocyte cultures were successfully established. EGF maximally stimulated keratocyte and epithelial cells at 25 ng/mL and 5 ng/mL, respectively. PDGF‐BB maximally stimulated keratocytes and epithelial cells at 50 ng/mL and 5 ng/mL, respectively. TGF‐β1 inhibited keratocytes at 5 ng/mL and 10 ng/mL, and epithelial cells at 1 ng/mL and 2 ng/mL. Conclusions Methods were established to maintain epithelial cells and keratocytes in vitro. PDGF‐BB and EGF stimulate, while TGF‐β1 inhibits the proliferation of epithelial cells and keratocytes. These growth factors may play a role in maintenance and repair of the equine cornea.     

Evaluation of formalin-fixed paraffin-embedded tissues from feline vaccine site-associated sarcomas for feline foamy virus DNA

OBJECTIVE: To evaluate a group of vaccine site-associated sarcomas (VSS) for the presence of feline foamy virus (FeFV) DNA, using polymerase chain reaction (PCR) methods. SAMPLE POPULATION: 50 formalin-fixed paraffin-embedded (FFPE) tissue blocks from VSS of cats. PROCEDURE: DNA was extracted from FFPE sections of each tumor, and regions of the gag and pol genes of FeFV were amplified by use of PCR methods, using 1 primer set for each region. Sensitivity of the method was compared between fresh and FFPE cells, using mouse kidney tissue that was injected with FeFV-infected cultured cells and using agarose-cell pellets. Results-Feline foamy virus DNA was not detected in VSS tissues. Sensitivity of the method was 10 times greater in fresh versus FFPE mouse tissues. Sensitivity of the method in fresh FeFV-infected cultured cells versus FFPE agarose-cell pellets was equal when fixation was 24 or 48 hours and 10 times greater when fixation was 72 hours or 1 week. CONCLUSIONS AND CLINICAL RELEVANCE: A PCR-based method can be successfully applied to FFPE tissues for FeFV DNA detection. Results suggest there is no direct FeFV involvement in the pathogenesis of VSS in cats.     

MAGNETIC RESONANCE IMAGING CHARACTERISTICS OF THE FOOT IN HORSES WITH PALMAR FOOT PAIN AND CONTROL HORSES

Palmar foot pain is a common cause of lameness. Magnetic resonance imaging (MRI) has the potential to detect damage in all tissues of the equine foot, but an understanding of the differences in magnetic resonance (MR) images between feet from horses with and without palmar foot pain is required. This study aimed to describe MR findings in feet from horses with no history of foot-related lameness, and to compare these with MR findings in horses with lameness improved by palmar digital local analgesia. Thirty-four limbs from horses euthanized with a clinical diagnosis of navicular syndrome (ameness >2 months duration, positive response to palmar digital nerve blocks and absence of other forelimb problems) (Group L), and 25 feet from age-matched horses with no history of foot pain (Group N) were examined. For each anatomic structure, MR signal intensity and homogeneity, size, definition of margins, and relationships with other structures were described. Alterations in MR signal intensity and homogeneity were graded as mild, moderate, or severe and compared between Groups L and N. Results revealed that there were significant differences in MR images between Groups N and L. Multiple moderate-severe MR signal changes were present in 91% of limbs from Group L and moderate (none were graded severe) in 27% of limbs from Group N. In most Group L limbs, more than three structures and frequently six to eight structures were abnormal. Concomitant abnormalities involved most frequently the deep digital flexor tendon, distal sesamoidean impar ligament, navicular bone, collateral sesamoidean ligament, and navicular bursa (with significant associations in severity grade between these structures), sometimes with involvement of the distal interphalangeal joint and/or its collateral ligaments. It was concluded that findings on MR images were different between horses with and without foot pain, and that pain localized to the foot was associated with MR changes in a variety of structures, indicating that damage to several structures may occur concurrently and that MR imaging was useful for evaluation of foot pain.     

Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs

MicroRNAs (miRNAs) are a class of small non-coding RNAs with a critical role in development and environmental responses. Efficient and reliable detection of miRNAs is an essential step towards understanding their roles in specific cells and tissues. However, gel-based assays currently used to detect miRNAs are very limited in terms of throughput, sensitivity and specificity. Here we provide protocols for detection and quantification of miRNAs by RT-PCR. We describe an end-point and real-time looped RT-PCR procedure and demonstrate detection of miRNAs from as little as 20 pg of plant tissue total RNA and from total RNA isolated from as little as 0.1 μl of phloem sap. In addition, we have developed an alternative real-time PCR assay that can further improve specificity when detecting low abundant miRNAs. Using this assay, we have demonstrated that miRNAs are differentially expressed in the phloem sap and the surrounding vascular tissue. This method enables fast, sensitive and specific miRNA expression profiling and is suitable for facilitation of high-throughput detection and quantification of miRNA expression.     

Litterarische Berichte

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Litterarische Berichte

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Art, Anteil und Verteilung niedrig molekularer Kohlenhydrate in der Rotbuche (Fagus sylvatica Linn)

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