Expression of porcine endogenous retroviruses (PERVs) in melanomas of Munich miniature swine (MMS) Troll

Porcine endogenous retroviruses (PERVs) are integrated in the genome of all pig breeds. Since some of them are able to infect human cells, they might represent a risk for xenotransplantation using pig cells or organs. However, the expression and biological role of PERVs in healthy pigs as well as in porcine tumours is largely unknown. Since we and others have recently shown overexpression of a human endogenous retrovirus, HERV-K, in human melanomas, we studied the expression of PERVs in melanomas of selectively bred Munich miniature swine (MMS) Troll. This breeding herd of MMS Troll is characterised by a high prevalence of melanomas, which histologically resemble various types of cutaneous melanomas in humans. Several genetic factors have been defined when studying inheritance of melanomas and melanocytic nevi in MMS Troll. Here we show that the polytropic PERV-A and PERV-B as well as the ecotropic PERV-C are present in the genome of all melanoma bearing MMS Troll investigated. Most interestingly, in the spleen, but not in other organs, recombinant PERV-A/C proviruses were found. PERV expression was found elevated in melanomas when compared to normal skin and viral proteins were expressed in melanomas and pulmonary metastasis-derived melanoma cell cultures. During passaging of these cells in vitro the expression of PERV mRNA and protein increased and virus particles were released as shown by RT activity in the supernatant and by electron microscopy. Genomic RNA of PERV-A, -B and -C were found in pelleted virus particles. Although PERV expression was elevated in melanomas and pulmonary metastasis-derived cell cultures, the function of the virus in tumour development is still unclear.     

Bacteraemia in 66 cats and antimicrobial susceptibility of the isolates (1995-2004)

Bacterial blood culture results of 292 privately owned cats presented to the Clinic for Small Animal Medicine, Ludwig Maximilian University Munich with signs of sepsis were evaluated retrospectively. Of the blood cultures, 23% were positive. In 88%, a single bacterial species was isolated. Of all bacterial isolates, 45% were Gram-positive, 43% were Gram-negative, and 12% were obligate anaerobes. The most frequently isolated bacteria were Enterobacteriaceae, obligate anaerobic species, Staphylococcus species and Streptococcus species. Of the cats with positive blood cultures, 32% were pretreated with antibiotics. Of all bacterial isolates, 77% were susceptible to enrofloxacin, 69% to chloramphenicol, 67% to gentamicin, and 64% to amoxycillin clavulanic acid. Only enrofloxacin reached an in vitro efficacy of more than 70% against Gram-positive and more than 74% against Gram-negative bacteria.     

Pedogenic carbonate recrystallization assessed by isotopic labeling: a comparison of 13C and 14C tracers

The C isotopic composition (δ¹³C) of pedogenic carbonates reflects the photosynthetic pathway of the predominant local vegetation because pedogenic (secondary) CaCO₃ is formed in isotopic equilibrium with soil CO₂ released by root and rhizomicrobial respiration. Numerous studies show the importance of pedogenic carbonates as a tool for reconstructing paleoecological conditions in arid and semiarid regions. The methodological resolution of these studies strongly depends on the time scale of pedogenic carbonate formation, which remains unknown. The initial formation rate can be assessed by ¹⁴C labeling of plants grown on loess and subsequent incorporation of ¹⁴C from rhizosphere CO₂ into newly formed carbonate by recrystallization of loess CaCO₃. We tested the feasibility of ¹⁴C and ¹³C tracers for estimating CaCO₃ recrystallization rates by simultaneous ¹⁴C and ¹³C labeling and comparison with literature data. ¹⁴C labeling was more efficient and precise in assessing recrystallization rates than ¹³C labeling. This is connected with higher sensitivity of ¹⁴C liquid scintillation counting when compared with δ¹³C measurement by IRMS. Further, assessment of very low amounts of incorporated tracer is more precise with low background signal (natural abundance), which is true for ¹⁴C, but is rather high for ¹³C. Together, we obtained better reproducibility, higher methodological precision, and better plausibility of recrystallization rates calculated based on ¹⁴C labeling. Periods for complete CaCO₃ recrystallization, extrapolated from rates based on ¹⁴C labeling, ranged from 130 (125–140) to 240 (225–255) y, while it was ≈ 600 (365–1600) y based on the ¹³C approach. In terms of magnitude, data from late‐Holocene soil profiles of known age provide better fit with modeled recrystallization periods based on the ¹⁴C approach.     

Discrimination of ‘San Marzano’ accessions: A comparison of minisatellite, CAPS and SSR markers in relation to morphological traits

‘San Marzano’ (SM) is one of the most widely known tomato (Solanum lycopersicum L.) cultivars, and is a classic example of a local variety with a premium value. Unfortunately, the original cultivated form is underrepresented in the Protected Denomination of Origin (PDO) area because of the incidence of contaminant and phenotypically similar genotypes. Our aim was to examine the ability of three DNA marker systems (minisatellite, cleaved amplified polymorphic sequence (CAPS) and simple sequence repeat (SSR)) to reveal the genetic diversity of tomato accessions that were, based on a morphological analysis, very similar. The data indicate that both minisatellites and SSRs can be used to genetically distinguish the analysed materials. Furthermore, these two marker systems depict relationships consistent with the hierarchal pattern obtained by the morphological data. As locally cultivated tomato accessions are often characterised by some degree of genetic variability, our results will be valuable in facilitating the purification, management and breeding of tomato germplasms. The differences between the marker systems employed are also discussed in relation to their usefulness in the agro-food chain.     

Artificial neural networks for estimating soil hydraulic parameters from dynamic flow experiments

Inverse methods are often used for estimating soil hydraulic parameters from experiments on flow of water through soil. We propose here an alternative method using neural networks. We teach a problem‐adapted network of radial basis functions (RBF) the relationship between soil parameters and transient flow patterns using a numerical flow model. The trained RBF network accurately identifies soil parameters from flow patterns not contained in the training scenarios. A comparison with the inverse method (Annealing‐Simplex) reveals a similarly good prediction by both approaches for randomly perturbed data and data from the real world. Nonetheless, the inverse method showed dependency on initial parameter estimates not required by the RBF network. Training demands moderately more computation and manpower than the inverse technique, but the absolutely stable and simple network application requires negligible resources. Thus, for individual applications, the network approach is slightly surpassed by the Annealing‐Simplex method. However, the RBF network has to be trained only once and, subsequently, it can be applied easily and without effort upon any number of laboratory experiments with standardized experimental setups.     

Microtubule stabilization reduces scarring and causes axon regeneration after spinal cord injury

Hypertrophic scarring and poor intrinsic axon growth capacity constitute major obstacles for spinal cord repair. These processes are tightly regulated by microtubule dynamics. Here, moderate microtubule stabilization decreased scar formation after spinal cord injury in rodents through various cellular mechanisms, including dampening of transforming growth factor-β signaling. It prevented accumulation of chondroitin sulfate proteoglycans and rendered the lesion site permissive for axon regeneration of growth-competent sensory neurons. Microtubule stabilization also promoted growth of central nervous system axons of the Raphe-spinal tract and led to functional improvement. Thus, microtubule stabilization reduces fibrotic scarring and enhances the capacity of axons to grow.