Effects of timing of soil frost thawing on Scots pine

Effects of the timing of soil thawing in the spring on Scots pine (Pinus sylvestris L.) were studied under controlled laboratory conditions. Sixteen 6-year-old saplings were lifted from the field, replanted in containers and placed in four treatments in controlled environment (CE) chambers with four replicate saplings per chamber. The saplings were held in the CE chambers during one simulated winter and one simulated growing season. The soil was frozen to -2 degrees C during a second simulated winter in the CE chambers, and the soil thawing treatments began at the end of the second simulated winter. Soil thawing began at various times before (no delay in thawing) and after (delay in thawing) chamber air conditions were changed from simulated winter to simulated summer. Delayed soil thawing subjected saplings to stress, with the severity of stress depending on the length of the delay in thawing. If there was no delay or only a short delay in soil thawing, stress was minor and reversible. A 2-week delay in soil thawing led to death of the saplings. Stress was apparent as decreases in the variable to maximal chlorophyll fluorescence ratio (Fv/Fm), chlorophyll a/b ratio and needle water potential. In needles of stressed saplings, apoplastic electrical resistance first decreased and then increased and there were anomalies in the electrical impedance spectra of the stems. Stress from the soil thawing treatments affected both root and shoot growth.     

Boron retranslocation in Scots pine and Norway spruce

We previously traced 10B-enriched boric acid from shoots to roots to demonstrate the translocation of boron (B) in Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.) seedlings. To gain a more detailed understanding of B translocation, we sought: (1) to demonstrate B retranslocation directly, by showing that foliar-applied 10B is located in the new growth after dormancy; and (2) to assess whether shoot-applied B affects growth in the long term. We applied 10B-enriched boric acid to needles of Scots pine and Norway spruce seedlings. After a dormancy period and 9 weeks of growth, small but significant increases in the 10B isotope were found in the new stem and needles of both species. In Scots pine, the total B concentration of the new stem was also increased. Both species contained polyols, particularly pinitol and inositol. Boron-polyol complexes may provide a mechanism for mobilizing B in these species. To determine the long-term effects of applied B, seedlings were grown for two growing seasons after the application of 10B to shoots. In Norway spruce, the proportion of 10B in the root systems and current needles of the harvest year was slightly higher than in the controls, and in Scots pine root systems, marginally so. The B treatment had no effect on growth of Norway spruce seedlings. In Scots pine seedlings, the B treatment caused a 33% increase in total dry mass and significantly increased the number of side branches.     

Effects of soil temperature on shoot and root growth and nutrient uptake of 5-year-old Norway spruce seedlings

Soil temperature is a main factor limiting root growth in the boreal forest. To simulate the possible soil-warming effect of future climate change, 5-year-old Norway spruce (Picea abies (L.) Karst.) seedlings were subjected to three simulated growing seasons in controlled environment rooms. The seedlings were acclimated to a soil temperature of 16 degrees C during the first (GS I) and third growing seasons (GS III), but were assigned to random soil-temperature treatments of 9, 13, 18 and 21 degrees C during the second growing season (GS II). In GS II, shoot diameter growth was lowest in the 21 degrees C treatment and root growth was lowest in the 9 degrees C treatment. In GS III, shoot height and root length growth improved in seedlings that had been kept at 9 degrees C during GS II, indicating compensatory growth in response to increased soil temperature. The temporary decrease in soil temperature had no long-lasting significant effect on seedling biomass or total nutrient uptake. At the end of GS III, fine roots of seedlings exposed to a soil temperature of 21 degrees C in GS II were distributed more evenly between the organic and mineral soil layers than roots of seedlings in the other treatments. During GS II and GS III, root growth started earlier than shoot growth, decreased during the rapid shoot elongation phase and increased again as shoot growth decreased.     

Concurrent bartonellosis and babesiosis in a dog with persistent thrombocytopenia

A 12-year-old castrated male West Highland White Terrier was referred because of recurrent episodes of collapsing. The dog was mildly anemic and severely thrombocytopenic and had high serum alanine aminotransferase activity. Infection with Bartonella vinsonii (berkhoffii) was initially diagnosed on the basis of serologic testing. Despite treatment with a series of antimicrobials and prolonged use of immunosuppressive drugs, thrombocytopenia persisted. After 5 months of treatment, Babesia canis organisms were seen during examination of a direct blood smear. The dog was treated with imidocarb dipropionate for babesiosis, after which thrombocytopenia resolved, and administration of immunosuppressive drugs was discontinued. Retrospective review of blood smears failed to identify organisms; however, polymerase chain reaction (PCR) analysis of multiple stored blood samples obtained during the 5-month period of persistent thrombocytopenia identified DNA of B. canis vogeli. Babesiosis may cause persistent, unexplained thrombocytopenia in dogs that are not anemic. A PCR assay can facilitate a diagnosis of babesiosis when organisms are not evident or when serologic testing fails to detect Babesia-specific antibodies.     

FDG‐PET IN HEALTHY AND EPILEPTIC LAGOTTO ROMAGNOLO DOGS AND CHANGES IN BRAIN GLUCOSE UPTAKE WITH AGE

Regional cerebral metabolism and blood flow can be measured noninvasively with positron emission tomography (PET). 2‐[¹⁸F]fluoro‐2‐deoxy‐D‐glucose (FDG) widely serves as a PET tracer in human patients with epilepsy to identify the seizure focus. The goal of this prospective study was to determine whether juvenile or adult dogs with focal‐onset epilepsy exhibit abnormal cerebral glucose uptake interictally and whether glucose uptake changes with age. We used FDG‐PET to examine six Lagotto Romagnolo dogs with juvenile epilepsy, two dogs with adult‐onset epilepsy, and five control dogs of the same breed at different ages. Three researchers unaware of dog clinical status visually analyzed co‐registered PET and magnetic resonance imaging (MRI) images. Results of the visual PET analyses were compared with electroencephalography (EEG) results. In semiquantitative analysis, relative standard uptake values (SUV) of regions of interest (ROI) drawn to different brain regions were compared between epileptic and control dogs. Visual analysis revealed areas of hypometabolism interictally in five out of six dogs with juvenile epilepsy in the occipital, temporal, and parietal cortex. Changes in EEG occurred in three of these dogs in the same areas where PET showed cortical hypometabolism. Visual analysis showed no abnormalities in cerebral glucose uptake in dogs with adult‐onset epilepsy. Semiquantitative analysis detected no differences between epileptic and control dogs. This result emphasizes the importance of visual analysis in FDG‐PET studies of epileptic dogs. A change in glucose uptake was also detected with age. Glucose uptake values increased between dog ages of 8 and 28 weeks and then remained constant.     

CEREBRAL GLUCOSE UTILIZATION MEASURED WITH HIGH RESOLUTION POSITRON EMISSION TOMOGRAPHY IN EPILEPTIC FINNISH SPITZ DOGS AND HEALTHY DOGS

In human epileptic patients, changes in cerebral glucose utilization can be detected 2‐deoxy‐2‐[¹⁸F] fluoro‐d ‐glucose positron emission tomography (FDG‐PET). The purpose of this prospective study was to determine whether epileptic dogs might show similar findings. Eleven Finnish Spitz dogs with focal idiopathic epilepsy and six healthy dogs were included. Dogs were examined using electroencephalography (EEG) and FDG‐PET, with epileptic dogs being evaluated during the interictal period. Visual and semi‐quantitative assessment methods of FDG‐PET were compared and contrasted with EEG findings. Three independent observers, unaware of dog clinical status, detected FDG‐PET uptake abnormalities in 9/11 epileptic (82%), and 4/8 healthy dogs (50%). Occipital cortex findings were significantly associated with epileptic status (P = 0.013). Epileptic dogs had significantly lower standardized uptake values (SUVs) in numerous cortical regions, the cerebellum, and the hippocampus compared to the control dogs. The lowest SUVs were found in the occipital lobe. White matter normalized and left‐right asymmetry index values for all pairs of homologous regions did not differ between groups. Visual evaluation of the EEGs was less sensitive (36%) than FDG‐PET. Both diagnostic tests were consensual and specific (100%) for occipital findings, but EEG had a lower sensitivity for detecting lateralized foci than FDG‐PET. Findings supported the use of FDG‐PET as a diagnostic test for dogs with suspected idiopathic epilepsy. Visual and semiquantitative analyses of FDG‐PET scans provided complementary information. Findings also supported the theory that epileptogenesis may occur in multiple brain regions in Finnish Spitz dogs with idiopathic epilepsy.     

Determination of semivolatile compounds in Baltic herring (Clupea harengus membras) by supercritical fluid extraction-supercritical fluid chromatography-gas chromatography-mass spectrometry

The on-line supercritical fluid extraction-supercritical fluid chromatography-gas chromatography method was applied to the determination of volatile compounds of raw and baked Baltic herring (Clupea harengus membras). The analytes were extracted with supercritical carbon dioxide at 45 degrees C and 10 MPa pressure. After extraction, the volatiles and coeluted lipids were separated on-line using supercritical fluid chromatography and the volatile fraction was introduced directly into a gas chromatograph. In all, 30 compounds were identified from fish samples with mass spectrometry. The most abundant compounds in the fresh Baltic herrings were heptadecane and 1-heptadecene. When the fish were stored for 3-6 days at 6 degrees C, the total peak area of the volatiles began to increase and the proportions of short chain acids (acetic, propanoic, 2-methylpropanoic, and 3-methylbutanoic) also increased. After 8-9 days of storage, 3-methylbutanoic acid comprised about 36 and 40% of all volatiles in raw and baked herring, respectively.     

Effects of planting orientation and density of willows on biomass production and nutrient leaching

Willow buffer strips are a promising vegetated filter for the reduction of non-point source pollution from agricultural land to watercourses resulting from the rapid growth of shoots and from the large amount and distribution of roots. A greenhouse experiment tested the hypothesis that a new planting method, planting cuttings horizontally, exhibited more biomass production and more effective filter than did planting cuttings vertically. Due to the different times taken for shoots break through the soil surface, the summed height of all shoots of horizontally planted cuttings was significantly smaller than that of the vertical ones at the beginning of the growth period. The difference in the height of all of the shoots between planting orientations decreased with time. After 16 weeks growth, the biomass of the stem, leaves and fine roots was not affected by the planting orientation. The coarse root biomass was larger in the horizontal treatments and decreased with increased density. The total biomass, as well as the biomass of stem or leaves, of each treatment, increased with increased planting density. The individual plant biomass, as well as that of the stem or leaves, decreased with increased planting density. Compared with the unplanted control treatment, the planted treatments significantly reduced the total-N, NO₃-N, PO₄-P and SO₄-S leaching. The planting orientation did not affect the nitrate leaching. The horizontally planted cuttings were slightly more effective for reducing the SO₄-S leaching and the vertically planted cuttings were slightly more effective for reducing the PO₄-P leaching. Lower PO₄-P leaching was observed only with higher planting density. With regard to the horizontal planting method, further studies are needed to explore the influence of different willow clones, the size of cuttings, pre-planting treatments, planting geometry (configuration) and soil conditions on survival, the number of shoots produced, the biomass production and the amount and distribution of roots.     

Electroencephalography findings in healthy and Finnish Spitz dogs with epilepsy: visual and background quantitative analysis

BACKGROUND: Qualitative and quantitative electroencephalography (EEG) parameters of healthy and Finnish Spitz dogs with epilepsy have not been determined. OBJECTIVE: To determine if EEG can provide specific characteristics to distinguish between healthy dogs and dogs with epilepsy. ANIMALS: Sixteen healthy and 15 Finnish Spitz dogs with epilepsy. METHODS: A prospective clinical EEG study performed under medetomidine sedation. Blinded visual and quantitative EEG analyses were performed and results were compared between study groups. RESULTS: Benign epileptiform transients of sleep and sleep spindles were a frequent finding in a majority of animals from both groups. The EEG analysis detected epileptiform activity in 3 Finnish Spitz dogs with epilepsy and in 1 healthy Finnish Spitz dog. Epileptiform activity was characterized by spikes, polyspikes, and spike slow wave complexes in posterior-occipital derivation in dogs with epilepsy and with midline spikes in control dog. The healthy dogs showed significantly less theta and beta activity than did the dogs with epilepsy (P < .01), but the only significant difference between healthy dogs and dogs with untreated epilepsy was in the alpha band (P < .001). Phenobarbital treatment increased alpha, beta (P < .001), and theta (P < .01), and decreased delta (P < .001) frequency bands compared with dogs with untreated epilepsy. CONCLUSIONS AND CLINICAL IMPORTANCE: Benign epileptiform transients of sleep could be easily misinterpreted as epileptiform activity. Epileptiform activity in Finnish Spitz dogs with epilepsy seems to originate from a posterior-occipital location. The EEG of dogs with epilepsy exhibited a significant difference in background frequency bands compared with the control dogs. Phenobarbital treatment markedly influenced all background activity bands. Quantitative EEG analysis, in addition to visual analysis, seems to be a useful tool in the examination of patients with epilepsy.     

Binding of GGA2 to the lysosomal enzyme sorting motif of the mannose 6-phosphate receptor

The GGAs are a multidomain protein family implicated in protein trafficking between the Golgi and endosomes. Here, the VHS domain of GGA2 was shown to bind to the acidic cluster-dileucine motif in the cytoplasmic tail of the cation-independent mannose 6-phosphate receptor (CI-MPR). Receptors with mutations in this motif were defective in lysosomal enzyme sorting. The hinge domain of GGA2 bound clathrin, suggesting that GGA2 could be a link between cargo molecules and clathrin-coated vesicle assembly. Thus, GGA2 binding to the CI-MPR is important for lysosomal enzyme targeting.