Functional characterization of post-junctional adrenergic receptor subtypes in bovine intra-mammary arteries

We examined the functional role of adrenergic receptor subtypes (ARs) in bovine intra-mammary arteries (IMAs), 1.5–2.5 mm internal diameter. Norepinephrine (NE) and phenylephrine (PE) produced concentration-dependent increases in tone in segments maintained at a previously determined optimal basal tension in vitro . The sensitivity of the tissue to NE and PE, based on -log molar ED_50s was 6.87 ± 0.17 and 7.05 ± 0.35, respectively. In addition a Schild analysis yielded antagonist affinities for the receptor mediating contractile responses to NE (pA2 value) of 10.46 ± 0.85 for prazosin and 6.29 ± 0.18 for yohimbine. These data indicate a dominance of functional alpha 1 (α₁) over alpha 2 (α₂)-ARs in this tissue. Based on the inhibitory effects of chloroethylclonidine (CEC) on PE responses and the further reduction in sensitivity when nifedipine was added to the CEC, also in the presence of PE, we conclude that there is more than one α₁-AR subtype, with a predominant role for α_1B-ARs in phenylephrine responses. Stimulation of beta (β)-ARs, resulted in relatively small reductions in tone (the highest magnitude of response was 25.94 ± 6.46% of the papaverine maximum at 3×10⁻⁶ M isoproterenol); in addition, propranolol did not significantly alter tissue sensitivity to NE. Additional characterization of functional autonomic receptor populations in this circulatory bed will form a basis for future studies on circulatory dynamics in the mammary gland.     

Chronological study of Mycoplasma hyopneumoniae infection, seroconversion and associated lung lesions in vaccinated and non-vaccinated pigs

A field trial was conducted to study Mycoplasma hyopneumoniae (Mh) infection dynamics by nested polymerase chain reaction (nPCR) and serology in pigs of a farm affected by enzootic pneumonia (EP). Moreover, correlation of Mh detection at different respiratory tract sites with presence of EP gross and microscopic lung lesions was assessed. These parameters were studied and compared between vaccinated (two doses at 1 and 3 weeks of age versus one dose at 6 weeks of age) and non-vaccinated pigs. Animals were monitored from birth to slaughter by nPCR from nasal swabs and by serology. From 3 to 22 weeks of age, an average of three pigs per treatment and per batch were necropsied (n = 302). The remaining pigs were sent to the slaughter (n = 103). Nasal, bronchial and tonsillar swabs were taken from the necropsied/slaughtered pigs; gross and microscopic EP-suggestive lung lesions were also assessed. Single and double vaccination resulted in earlier seroconversion and higher percentage of Mh seropositive pigs compared to control group. At slaughter, double vaccinated pigs showed lower percentage of EP-compatible gross lung lesions and lower Mh prevalence at upper respiratory tract sites (nasal cavity and tonsil) than control pigs.     

A new<Emphasis Type="Italic">Aphidius</Emphasis> species (Hymenoptera: Braconidae: Aphidiinae) from high-montane areas of southeastern Europe

Aphidius montenegrinus sp.n. from Serbia and Montenegro is described and illustrated. It parasitizedAcyrthosiphon daphnidis Ilharco, onDaphne alpina L. The parasitoid was described and illustrated using scanning electron microscope photographs and line drawing. The new species is a member of theAcyrthosiphon aphid parasitoid guild and was collected from Zabojsko Jezero Lake in Montenegro. The parasitoid and the hyperparasitoid spectrum ofA. daphnidis are presented for the first time. http://www.phytoparasitica.org posting April 30, 2004.     

Chemical authentication of extra virgin olive oil varieties by supervised chemometric procedures.

This work has focused on discriminating extra virgin olive oils from Sabina (Lazio, Italy) by olive fruit variety (cultivar). A set of oils from five of the most widespread cultivars (Carboncella, Frantoio, Leccino, Moraiolo, and Pendolino) in this geographical area was analyzed for chemical composition using only the Official Analytical Methods, recognized for the quality control and commercial classification of this product. The obtained data set was converted into a computer-compatible format, and principal component analysis (PCA) and a method based on the Fisher F ratio were used to reduce the number of variables without a significant loss of chemical information. Then, to differentiate these samples, two supervised chemometric procedures were applied to process the experimental data: linear discriminant analysis (LDA) and artificial neural network (ANN) using the back-propagation algorithm. It was found that both of these techniques were able to generalize and correctly predict all of the samples in the test set. However, these results were obtained using 10 variables for LDA and 6 (the major fatty acid percentages, determined by a single gas chromatogram) for ANN, which, in this case, appears to provide a better prediction ability and a simpler chemical analysis. Finally, it is pointed out that, to achieve the correct authentication of all samples, the selected training set must be representative of the whole data set.     

A mathematical comparison of classification structures: The case of the USDA Soil Taxonomy

Biologists have not paid much attention to the mathematical structures of biological classifications and their interpretations. A few have made such an analysis mainly using statistical distribution models. Comparisons with nonbiological classifications are needed to know if such structures are the result of biological evolution or of taxonomic practices, utilitarian bias, and/or subconscious cognitive rules. We compare the biological classification of a target group of soil borne plant parasitic nematodes (the Tylenchina suborder) with a non-biological one (the USDA 8th edition of the Keys to Soil Taxonomy). The authors made use of the same tools used in previous papers by biologists introducing other classifications. The results show that both taxonomies are information systems that try to optimize the information flow and fit well to the same distribution models. The analysis does not show any idiosyncrasies of biological classifications with respect to pedological ones, thus, supporting the idea that these products are the result of subconscious cognitive rules used by humankind to classify the world. Published in Russian in Pochvovedenie, 2006, No. 7, pp. 795–803. The text was submitted by the authors in English.     

Use of stem diameter variations to detect plant water stress in tomato

The sensitivity of stem diameter variations (SDV) measured with linear variable transducer (LVDT) sensors as indicators of plant water status in tomato was evaluated. Two tomato crops were grown sequentially in a sandy loam soil in an unheated plastic greenhouse. These were an autumn–winter tomato crop (autumn crop) and a spring–summer tomato crop (spring crop). One drying cycle of 61 days was imposed to the autumn crop in winter at 92 days after transplanting (DAT). Two drying cycles, each of 29 days, were applied to the spring crop, to young (58 DAT) and mature plants (121 DAT). For each drying cycle, four replicate plots did not receive irrigation, and four were well watered. During each drying cycle, LVDT sensors continuously measured SDV, and daily measurements were made of leaf (Ψ _leaf) and stem water potential (Ψ _stem). SDV data was interpreted using the SDV-derived indices, maximum daily shrinkage (MDS) and stem growth rate (SGR). The response of SDV-derived indices to water deficit differed with (1) climatic conditions during stress imposition and (2) crop age. In the winter drying cycle of the autumn crop, the responses of the SDV-derived indices to soil drying were relatively small and slower than Ψ _leaf and Ψ _stem. Under warmer conditions, the SDV-derived indices were much more responsive to soil drying. In rapidly growing young plants, where SDV was characterized by high SGR and small MDS, SGR was the most sensitive SDV-derived index. In more mature plants with little stem growth, MDS was the most sensitive SDV-derived index. In mature plants grown in warm to hot conditions, MDS (1) responded at a similar time or earlier than Ψ _leaf and Ψ _stem and (2) had larger “signal” values (ratio of values from unwatered to control plants) than Ψ _leaf and Ψ _stem. However, there was appreciably more “noise” (coefficient of variation, CV) associated with the SDV-derived indices, giving lower “sensitivity” values, determined from “signal” to “noise” ratios, than for Ψ _leaf and Ψ _stem. Regression analysis between MDS of well-watered plants and climatic variables gave best results for a linear relationship between MDS and daily maximum vapor pressure deficit. There were strong linear relationships between MDS and Ψ _leaf for each drying cycle. The slopes of these relationships differed with crop age indicating that there was no constant relationship between MDS and Ψ _leaf for a whole season. Overall these results demonstrated that MDS and SGR can be sensitive indicators of the water status of tomato crops under conditions of moderate to high evaporative demand. However, the variability associated with the SDV-derived indices and the changing MDS–Ψ _leaf relationship with crop age represent major issues regarding the development of irrigation scheduling protocols for tomato.     

Convergence of developmental mutants into a single tomato model system: 'Micro-Tom' as an effective toolkit for plant development research

Background

The tomato (Solanum lycopersicum L.) plant is both an economically important food crop and an ideal dicot model to investigate various physiological phenomena not possible in Arabidopsis thaliana. Due to the great diversity of tomato cultivars used by the research community, it is often difficult to reliably compare phenotypes. The lack of tomato developmental mutants in a single genetic background prevents the stacking of mutations to facilitate analysis of double and multiple mutants, often required for elucidating developmental pathways.

Results

We took advantage of the small size and rapid life cycle of the tomato cultivar Micro-Tom (MT) to create near-isogenic lines (NILs) by introgressing a suite of hormonal and photomorphogenetic mutations (altered sensitivity or endogenous levels of auxin, ethylene, abscisic acid, gibberellin, brassinosteroid, and light response) into this genetic background. To demonstrate the usefulness of this collection, we compared developmental traits between the produced NILs. All expected mutant phenotypes were expressed in the NILs. We also created NILs harboring the wild type alleles for dwarf, self-pruning and uniform fruit, which are mutations characteristic of MT. This amplified both the applications of the mutant collection presented here and of MT as a genetic model system.

Conclusions

The community resource presented here is a useful toolkit for plant research, particularly for future studies in plant development, which will require the simultaneous observation of the effect of various hormones, signaling pathways and crosstalk.     

Segmentation of 3D images of plant tissues at multiple scales using the level set method

Background

Developmental biology has made great strides in recent years towards the quantification of cellular properties during development. This requires tissues to be imaged and segmented to generate computerised versions that can be easily analysed. In this context, one of the principal technical challenges remains the faithful detection of cellular contours, principally due to variations in image intensity throughout the tissue. Watershed segmentation methods are especially vulnerable to these variations, generating multiple errors due notably to the incorrect detection of the outer surface of the tissue.

Results

We use the level set method (LSM) to improve the accuracy of the watershed segmentation in different ways. First, we detect the outer surface of the tissue, reducing the impact of low and variable contrast at the surface during imaging. Second, we demonstrate a new edge function for a level set, based on second order derivatives of the image, to segment individual cells. Finally, we also show that the LSM can be used to segment nuclei within the tissue.

Conclusion

The watershed segmentation of the outer cell layer is demonstrably improved when coupled with the LSM-based surface detection step. The tool can also be used to improve watershed segmentation at cell-scale, as well as to segment nuclei within a tissue. The improved segmentation increases the quality of analysis, and the surface detected by our algorithm may be used to calculate local curvature or adapted for other uses, such as mathematical simulations.

     

Purification and properties of a neutral peroxidase isozyme from turnip (Brassica napus L. Var. Purple Top White Globe) roots

A neutral peroxidase isozyme (pI 7.2) from turnip roots (TNP) was purified to homogeneity and partially characterized. TNP is a monomeric glycoprotein with 9.1% carbohydrate content and a molecular weight of 36 kDa. Optimum pH values for activity using 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) and guaiacol as H donors were 4.5 and 5.5, whereas the K(m) values were 0.7 and 3.7 mM, respectively. The ABTS K(m) was approximately 7 times higher than that reported for basic commercial horseradish peroxidase (HRP-C). TNP retained approximately 70% activity after 11 min of heating at 65 degrees C, whereas the activation energy for inactivation (132 kJ/mol) was higher than or comparable to that of other peroxidases. The low ABTS K(m) and high specific activity (1930 units/mg) gave a high catalytic efficiency (500 M(-1) s(-1)). These properties make TNP an enzyme with a high potential as an alternative to HRP in various applications.