Insights into the molecular mechanism of tolerance to carboxylic acid amide (CAA) fungicides in Pythium aphanidermatum

BACKGROUND: Tolerance to the oomycete‐specific carboxylic acid amide (CAA) fungicides is a poorly understood mechanism in Pythium species. The root‐rot and damping‐off causative agent Pythium aphanidermatum and the CAA fungicide mandipropamid (MPD) were used to investigate the molecular basis of CAA tolerance. RESULTS: Five genes putatively involved in carbohydrate synthesis were identified and characterised: one chitin synthase gene, PaChs, and four cellulose synthase genes PaCesA1 to PaCesA4, of which PaCesA3 encodes the MPD target enzyme. These genes were differentially expressed throughout the life cycle of P. aphanidermatum. Mycelium treated with MPD concentrations slightly affecting mycelial growth did not cause a change in PaCesA3 expression nor a strong upregulation of PaCesA homologues. The high tolerance level of P. aphanidermatum and the lack of PaCesA upregulation imply that MPD tolerance is the result of a specific amino acid configuration in the cellulose synthase 3 (CesA3) target enzyme. Indeed, P. aphanidermatum displays the amino acid L1109 which is also associated with MPD resistance in artificial mutants of Phytophthora species. CONCLUSION: It is concluded that MPD tolerance in P. aphanidermatum is not caused by compensatory mechanisms but most likely by an inherent target‐site configuration in PaCesA3 that hinders MPD binding to the enzyme pocket. Copyright © 2012 Society of Chemical Industry     

Thoracolumbar dorsolateral laminectomy with osteotomy of the spinous process in fourteen dogs

Objective— To describe outcome after an alternative unilateral approach to the thoracolumbar spine for dorsal laminectomy.
Study Design— Retrospective clinical study.
Animals— Dogs (n=14) with thoracolumbar spinal cord compression.
Methods— Thoracolumbar spinal cord compression was lateral (6 dogs), dorsal (4), and dorsolateral (4) caused by subarachnoid (7) and synovial cysts (2) and intradural-extramedullary neoplasia (5). All dogs were treated by dorsal laminectomy with osteotomy of the spinous process using a unilateral paramedian approach. The contralateral paraspinal muscles were not stripped from the spinous process and the osteoligamentous complexes were preserved. Retraction of the spinous process and muscles to the contralateral side resulted in complete visualization of the dorsal vertebral arch thereby allowing dorsal laminectomy to be performed.
Results— No technique complications occurred. Approximately 75% exposure of the spinal cord (dorsal and lateral compartments) was achieved providing adequate visualization and treatment of the lesions. Transient deterioration of neurologic state occurred in 5 dogs because of extensive spinal cord manipulation. At long-term follow-up, 6 dogs were normal, 6 had clinical improvement, and 2 were unchanged.
Conclusion— Dorsal laminectomy after osteotomy and retraction of the spinous process may be considered in canine patients with dorsal, dorsolateral, or lateral compression to facilitate adequate decompression of the spinal cord.
Clinical Significance— This surgical technique offers an alternative approach to the thoracolumbar spine and spinal cord by a modified dorsal laminectomy that preserves the paraspinal muscle integrity on the contralateral side.     

The crystal structure of [Fe]-hydrogenase reveals the geometry of the active site

Biological formation and consumption of molecular hydrogen (H2) are catalyzed by hydrogenases, of which three phylogenetically unrelated types are known: [NiFe]-hydrogenases, [FeFe]-hydrogenases, and [Fe]-hydrogenase. We present a crystal structure of [Fe]-hydrogenase at 1.75 angstrom resolution, showing a mononuclear iron coordinated by the sulfur of cysteine 176, two carbon monoxide (CO) molecules, and the sp2-hybridized nitrogen of a 2-pyridinol compound with back-bonding properties similar to those of cyanide. The three-dimensional arrangement of the ligands is similar to that of thiolate, CO, and cyanide ligated to the low-spin iron in binuclear [NiFe]- and [FeFe]-hydrogenases, although the enzymes have evolved independently and the CO and cyanide ligands are not found in any other metalloenzyme. The related iron ligation pattern of hydrogenases exemplifies convergent evolution and presumably plays an essential role in H2 activation. This finding may stimulate the ongoing synthesis of catalysts that could substitute for platinum in applications such as fuel cells.     

Storm damage of Douglas-fir unexpectedly high compared to Norway spruce

Context

Since storm damage has a large impact on forest management in Central Europe, we investigated the main storm risk factors for two important conifer species, Douglas-fir (Pseudotsuga menziesii [Mirbel] Franco) and Norway spruce (Picea abies [L.] Karst.).

Aims

We compared general storm damage levels of Douglas-fir and Norway spruce, the latter being known to have high storm risk among European tree species.

Methods

Generalized linear mixed models and boosted regression trees were applied to recorded storm damage of individual trees from long-term experimental plots in southwest Germany. This included two major winter storm events in 1990 and 1999. Over 40 candidate predictors were tested for their explanatory power for storm damage and summarized into predictor categories for further interpretation.

Results

The two most important categories associated with storm damage were timber removals and topographic or site information, explaining between 18 and 54 % of storm damage risk, respectively. Remarkably, general damage levels were not different between Douglas-fir and Norway spruce.

Conclusion

Under current forest management approaches, Douglas-fir may be considered a species with high storm risk in Central Europe, comparable to that of Norway spruce.     

Salmonella in poultry flocks and humans--S. enterica subspecies enterica serovar Enteritidis in the past

After importing of breeder lines for laying flocks from Canada into the former GDR in 1966 the egg industry in this country was completely isolated from that in Western Germany or other Western European countries until opening the border in Germany in 1989. Because of this isolation from other countries, an analysis of the clonal diversity of Salmonella (S.) Enteritidis isolates originated from humans, chickens and food in the former GDR during the 1980s would provide a unique opportunity to obtain new insight into factors that may have triggered the S. Enteritidis epidemic. While isolates had previously been typed by the phage typing scheme of Lalko and Laszlo we applied for the first time the extended phage typing scheme by Ward for the retrospective analysis of the S. Enteritidis strains. Furthermore, isolates of phage type (PT) 4/6 (Ward / Lalko and Laszlo) from different livestocks were investigated by ribotyping. Although in total the PT4/6 dominated between 1986 and 1989 in poultry, other phage types have prevailed in the early 1980s and represented a considerable fraction of isolates until 1989. For instance, PT8/7 was isolated from one large layer farm (district Halle) from 1988 until 1989. During that time in another farm (district Cottbus) only PT8/7 was detected too. PT4/6 was isolated from neither of these two laying hen farms. The strains of PT4/6 could be distinguished by ribotyping in 19 different subtypes. The strains from the northern farms were distinct from those isolated in the southern regions. As farms which were harbouring either PT4/6 or PT8/7 had obtained laying hens from the same sources (breeder lines in Deersheim and Spreenhagen) it is highly probable that S. Enteritidis infection was acquired from the environment at each individual farm. This conclusion is also supported by the presence of different PT4/6 ribotypes in different farms. The presence of different phage types or PT4/6 ribotypes at different farms of laying hens suggests that in each case the S. Enteritidis strains present in the environment were able to enter chicken flocks.