Validation of tissue microarrays for immunohistochemical analyses of canine lymphomas.

In most validation studies of tissue microarrays (TMAs), a fixed number of cores with a given diameter are analyzed to determine the degree of accuracy by which the TMA represents the whole section. The statistical model described in the present study predicts this property for various combinations of 2 core sizes (0.6 mm and 1.2 mm) and different core numbers. The model was based on artificial TMA core biopsies generated from Ki-67 and active caspase-3 immunostains of 40 canine lymphoma samples. Positivity was scored on a continuous scale, and a large number of cells were analyzed with the help of semiautomated cell counting. Despite considerable differences in range and distribution of Ki-67 and active caspase-3 positivity values, the model predictions showed a high degree of agreement for both markers. Comparison of 0.6 mm and 1.2 mm cores indicated that the use of small cores necessitates inclusion of a larger number of samples but requires counting a markedly smaller number of cells. Suitability of TMAs to determine the immunophenotype of the whole section was assessed using 2 different combinations of core sizes and numbers. Both displayed a high degree of concordance with the whole section (kappa(0.6) = 0.79; kappa(1.2) = 0.91). The present study provides a basis for the use of TMAs in future high-throughput immunohistochemical investigations of selected markers in canine lymphomas. The statistical model presented can be used to determine an optimal TMA design depending on a desired accuracy.     

Processing of remote sensing information to retrieve leaf area index in barley: a comparison of methods

Precision Agriculture - Leaf area index (LAI) is a key variable in understanding and modeling crop-environment interactions. With the advent of increasingly higher spatial resolution satellites and...     

Nitrogen mineralization from mature bio‐waste compost in vineyard soils. III Simulation of soil mineral‐nitrogen dynamics

A model for nitrogen (N) dynamics in compost‐amended vineyard soils was tested for its predictive power. A soil–mineral N data set from a 3‐year field study on four different vineyard sites was used for model evaluation. The soils were treated with mature bio‐waste compost (30 and 50 Mg ha^–1 fresh matter, respectively). The model calculated soil mineral‐N contents at all sites with an overall mean bias error of –2.2 kg N ha^–1 for layers of 0.1 m thickness and an overall mean absolute error of 7.4 kg N ha^–1 layer^–1. Modeling efficiencies for the simulations of the respective treatments ranged from –0.05 to 0.41, and Willmott's Index of Agreement showed values of between 0.41 and 0.81. Acceptable model predictions as defined by the observed variability of mineral‐N contents in the respective soils ranged from 40% to 72%. A strong increase in soil mineral‐N concentration following the compost application at all sites could not be reproduced with the model, thereby reducing the prediction accuracy significantly. The model performance confirms that previously derived N‐mineralization parameters are suitable to describe the N release from soil‐applied mature bio‐waste compost under the environmental conditions of vineyards in Germany.     

Nitrogen mineralization from mature bio‐waste compost in vineyard soils. I. Long‐term laboratory incubation experiments

The steadily increasing utilization of bio‐waste compost in German viticulture requires a more detailed investigation of nitrogen (N) mineralization parameters for mature bio‐waste compost applied to vineyard soils. N mineralization kinetics were described with two superposing exponential equations. Long‐term aerobic laboratory incubation experiments of 12 soil‐compost substrates revealed that 5±2.8% of its total N content could be released from a rapidly decomposable fraction (half‐life period t₅₀ = 41 d at 15°C) and another 60±2.9% from a slower decomposable fraction (t₅₀ = 490 d). The remaining proportion (35%) is considered not to be released in the medium term. The obtained potentially mineralizable nitrogen of 65% of total compost N significantly differs from current fertilizer recommendations, which were adopted from calculations for agricultural conditions. For fertilizer recommendations in viticulture, we recommend the consideration of a higher N‐mineralization potential for organic fertilizers.     

Methylmercury production in the anoxic hypolimnion of a Dimictic Seepage Lake

Experimental results and field data indicated that methyl-Hg was produced within a layer of bacterioplankton near the top of the anoxic hypolimnion of Pallette Lake. In situ incubations at ambient Hg concentrations indicated that the net flux of methyl-Hg from the layer was between 50 and 100 pmol/m^2*d. This input was sufficient to account for the summer accumulation of methyl-Hg in the entire hypolimnion and it exceeded atmospheric inputs by 2 orders of magnitude. Maximum rates of net methylation occurred in the same region of the water column where we observed maximum rates of sulfate reduction. The measured rates were: 100 fmol rriethyl-Hg/L^*d and 90 nmol SO₄/L^*d. Sulfate reducing enrichment cultures isolated from the hypolimnion were also able to methylate Hg in the laboratory. Sulfate reduction did not occur in anoxic profundal sediments during summer and we infer from ancillary data that methylation in profundal sediments was also low. Whole-lake rates of sulfate reduction in the hypolimnetic layer and shallow sediments were roughly equivalent, but we cannot yet compare methylation rates at these sites due to large uncertainties in the littoral flux of methyl-Hg. We propose that zones of Hg methylation and SO4 reduction follow the oxic/anoxic boundary in both the watercolumn and sediments. The relative importance of watercolumn and sediment processes will depend on the physical and chemical structure of a given lake.     

Induction of B cell unresponsiveness to noninherited maternal HLA antigens during fetal life

Patients who have received many transfusions become highly sensitized and develop antibodies against almost all HLA alloantigens, so that finding a cross-match negative kidney donor is difficult. A survey of those patients showed that 50 percent did not form antibodies against the noninherited maternal HLA antigens. Apart from the obvious clinical implications, the data indicate that a human equivalent of murine neonatal or actively acquired tolerance has now been identified.     

Pandemic influenza is a zoonosis, as it requires introduction of avian-like gene segments in the human population

Human influenza viruses manage to cause epidemics almost every year. The circulating viruses change their surface glycoproteins by accumulating mutations (antigenic drift) which results in variant viruses of the same subtype that are able to evade the immune pressure in the population. Every now and then, a completely new subtype of influenza A virus is introduced in the human population, which can result in an influenza pandemic. Pandemic human influenza viruses have been emerging for many centuries. Based on the genetic information of influenza viruses that have been isolated in this century, introduction of genes of the avian influenza virus reservoir obviously is required. Interspecies transmission, via another mammalian host and reassortment of avian and human influenza viruses are potential mechanisms for such an introduction. A summary of the cases in which influenza viruses containing avian-like gene segments were introduced into the human population is presented. In three cases, such infections resulted in conjunctivitis. Influenza-like illness and even pneumonia was reported in some other infections. Finally, a mortality rate of 33% was observed in the avian influenza A (H5N1) viruses that infected 18 people in Hong Kong in 1997. Although some of these viruses fulfilled some criteria of pandemic influenza viruses, they lacked the ability to rapidly spread through the human population.     

Nitrogen mineralization from mature bio‐waste compost in vineyard soils II. Test of N‐mineralization parameters in a long‐term in situ incubation experiment

A field incubation experiment was carried out to test the applicability of N‐mineralization parameters for mature bio‐waste compost for use in a simulation model. The parameters were previously obtained from a laboratory experiment. Micro‐lysimeters were used for incubation, containing four different vineyard soils that were treated with three different compost‐application rates (0, 30, and 50 Mg compost ha^–1). Between 2.0% and 45.2% of total bio‐waste compost N was mineralized and leached from the micro‐lysimeters during the two‐year investigation period. The application of a simulation model for soil N dynamics revealed two major drawbacks of the model: (1) in most of the soils, extraordinary high mineralization rates were observed within a few weeks after compost amendment, which could not be explained by the model, and (2) the average compost‐N‐mineralization rates were estimated as being close to the observed rates (–6%), but distinct deviations in some cases (–46% to +29%) led to considerable miscalculations in long‐term simulations. Excluding the effect of these two processes from the data set, the remaining variance could be well explained by the model for all soils treated with compost (modeling efficiency ≥0.98). Based on the average performance, the mineralization parameters for mature bio‐waste compost are considered to be applicable for use in any simulation model based on the double‐exponential approach for calculating fertilizer recommendations, whereas the functions calculating the impact of environmental factors on N mineralization in the model need to be revised. The initial mineralization flush observed in most of the compost treatments was attributed to a priming effect. The experiment showed that such a priming effect can cause exceptionally high rates of N mineralization from mature bio‐waste compost in a viticultural environment, which exceed the potential mineralization rates known for bio‐waste compost applied to arable soils in Germany.     

The influence of allogeneic cells on the human T and B cell repertoire

Clinical transplantation is often complicated by rejection episodes, in which the immune system of the recipient reacts to the foreign transplantation (HLA) antigens on the graft. This immune response includes humoral and cellular components. In the first, B lymphocytes form antibodies to the HLA alloantigens. In the second, CD8+ T lymphocytes recognize and react to HLA class I antigens, and CD4+ T cells react to HLA class II antigens. The frequency and severity of these rejection episodes can be diminished by immunosuppressive drugs, HLA matching between donor and recipient, and immune modulation by blood transfusion. Effective HLA matching between donor and recipient is not always possible and often not necessary. Insight into the factors that influence the T and B cell repertoire after blood transfusion might lead to new approaches to improve graft survival.     

Anatomy and dynamics of a supramolecular membrane protein cluster

Most plasmalemmal proteins organize in submicrometer-sized clusters whose architecture and dynamics are still enigmatic. With syntaxin 1 as an example, we applied a combination of far-field optical nanoscopy, biochemistry, fluorescence recovery after photobleaching (FRAP) analysis, and simulations to show that clustering can be explained by self-organization based on simple physical principles. On average, the syntaxin clusters exhibit a diameter of 50 to 60 nanometers and contain 75 densely crowded syntaxins that dynamically exchange with freely diffusing molecules. Self-association depends on weak homophilic protein-protein interactions. Simulations suggest that clustering immobilizes and conformationally constrains the molecules. Moreover, a balance between self-association and crowding-induced steric repulsions is sufficient to explain both the size and dynamics of syntaxin clusters and likely of many oligomerizing membrane proteins that form supramolecular structures.