Characterization of Amaranthus cruentus L. seed proteins by 2-DE and LC/MS–MS: Identification and cloning of a novel late embryogenesis-abundant protein

Amaranth is taking great attention as an important cereal crop that could fulfill food requirements for the growing population, especially in developing countries. However, the protein composition of these seeds is not well known yet. We have used the proteomics tools to characterize amaranth seed proteome. About 400 proteins spots were resolved on two-dimensional gel electrophoresis (2-DE) and protein spots were analyzed by LC-MS/MS (liquid chromatography tandem mass spectrometry). Identified proteins were related to stress and defense responses, metabolic, respiratory, and oxide-reduction processes. One abundant spot was identified as a Late Embryogenesis Abundant (LEA) protein and the gene was cloned and characterized. The AcLEA cDNA contains a 418 bp open reading frame (ORF) encoding a polypeptide of 139 amino acids. Bioinformatics analysis showed that AcLEA belongs to LEAs Group 5. Proteomics is a powerful technique that could be used even in non-sequenced organisms such as amaranth. The obtained information reveals that amaranth seed, beyond the classical seed storage proteins, contains proteins related to protection against stress. The identification of these proteins opens the door to the application of new strategies to improve the quality of amaranth production.     

Fungal diversity in coffee plantation systems and in a tropical montane cloud forest in Veracruz,Mexico

We compared the abundance, species richness and diversity of saprobic filamentous microfungi in the forest and in coffee plantation systems (with different biophysical structures of the vegetation and agricultural management) and evaluated the degree of similarity in species composition among these sites. Soil washing was used to isolate the saprobic filamentous microfungi. Emerging colonies were quantified and transferred to tubes with culture medium and then mounted on semi-permanent slides for identification under the microscope. From 90 samples and 4,500 inoculated soils particles, 415 species were distinguished. The genera Trichoderma, Penicillium, Fusarium, Chaetomium and Humicola were the most frequent in all study sites. The transformation of the tropical montane cloud forest in coffee plantation had no significant effect on the abundance, species richness and diversity of the saprobic filamentous microfungi. Effects of the biophysical structures of the vegetation and the agricultural management of the sites were only detected at the level of dominant genera (Fusarium, Trichoderma and Penicillium) and on the evenness. The low degree of similarity among the six sites suggests the existence of a high exchange of species.     

A New Simple Method for the Evaluation of Mineral Elements in Different Oilseeds

A method for the analysis of seven mineral contents—that is, potassium (K), calcium (Ca), phosphorus (P), iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu)—was investigated in some oilseeds like flax, sesame, pumpkin, quinoa, and yellow mustard. X-ray Spectral Analysis method with Spectrometer EDX-900HS was used for the analytical concentrations of the macro and micro elements of the oilseeds. A relatively high level of Zn content was found in sesame and pumpkin seeds, while the highest level of Fe content was found in mustard seeds. Regarding the macro elements (K, Ca, and P) content, the highest levels of concentration were found in flax and mustard seeds. The highest values of Cu were obtained in sesame seeds while those of Mn were in pumpkin and brown flax seeds. The results showed that the method is a reliable and simple analytical procedure to characterize mineral elements in oilseeds with a very high analytical performance.     

Effect of tillage and N fertilization on microbial physiological profile of soils cultivated with wheat

Microbial diversity might be a good indicator of ecosystem disturbance and functioning. We determined the effect of soil disturbances, such as N fertilization and tillage management, on soil microbial communities in a Typic Argiudoll of the El Salado river basin (Argentina). Microbial activity and substrate utilization provided a metabolic fingerprint of the soil microbial community. Univariate and multivariate analyses were used to differentiate responses to N fertilization and tillage at each of three growth stages of wheat (Triticum aestivum L.). Tillage had an adverse effect on microbial diversity, in which reduced and conventional tillage treated soils had different populations. However, N fertilization also altered microbial diversity depending on the crop developmental stage considered. Metabolism of carboxylic acids and carbohydrates were the main indicators of functional microbial activity and diversity. Although the substrate consumption profile of 32 C substrates did not provide insights into the fundamental ecological interactions that may induce changes in microbial population, it allowed us to demonstrate the alterations of microbial diversity as a result of tillage. We conclude that tillage and N fertilization altered microbial diversity.     

Systolic time intervals assessed by 2-D echocardiography and spectral Doppler in the horse

In the present study, the ‘acoustic windows’ for the measurement of the left ventricular systolic time intervals is studied by means of 2‐D echocardiography and cardiac Doppler ultrasonography, and the normal values in the horse (n = 112) are determined. The left ventricular isovolumetric contraction time, the pre‐ejection period (PEP), as well as the left ventricular ejection time (LVET) have been measured, and the values of the left ventricular total electromechanical systole (LVTES) and the PEP‐to‐LVET ratio have been calculated. It has been established that the most suitable window for the measurement of the aforementioned indices in 2‐D echocardiography is the right parasternal window in a view in short axis at the level of the cardiac base to measure the aortogram. In Doppler ultrasonography, the preferred window is the left parasternal using the five chambers apical view. The following values have been acquired: PEP = 0.071 ± 0.01 s; LVET = 0.532 ± 0.097 s; LVTES = 0.6 ± 0.1 s and PEP‐to‐LVET = 0.138 ± 0.025 measured by 2‐D echocardiography and PEP = 0.068 ± 0.009 s; LVET = 0.527 ± 0.076 s; LVTES = 0.598 ± 0.098 s and PEP‐to‐LVET = 0.131 ± 0.01 measured by Doppler ultrasonography.     

Neural encoding of olfactory recognition memory

Our work with both sheep and mouse models has revealed many of the neural substrates and signalling pathways involved in olfactory recognition memory in the main olfactory system. A distributed neural system is required for initial memory formation and its short-term retention-the olfactory bulb, piriform and entorhinal cortices and hippocampus. Following memory consolidation, after 8 h or so, only the olfactory bulb and piriform cortex appear to be important for effective recall. Similarly, whereas the glutamate-NMDA/AMPA receptor-nitric oxide (NO)-cyclic GMP signalling pathway is important for memory formation it is not involved in recall post-consolidation. Here, within the olfactory bulb, up-regulation of class 1 metabotropic glutamate receptors appears to maintain the enhanced sensitivity at the mitral to granule cell synapses required for effective memory recall. Recently we have investigated whether fluctuating sex hormone levels during the oestrous cycle modulate olfactory recognition memory and the different neural substrates and signalling pathways involved. These studies have used two robust models of social olfactory memory in the mouse which either involve social or non social odours (habituation-dishabituation and social transmission of food preference tasks). In both cases significant improvement of learning retention occurs when original learning takes place during the proestrus phase of the ovarian cycle. This is probably the result of oestrogen changes at this time since transgenic mice lacking functional expression of oestrogen receptors (ERalpha and ERbeta, the two main oestrogen receptor sub-types) have shown problems in social recognition. Therefore, oestrogen appears to act at the level of the olfactory bulb by modulating both noradrenaline and the glutamate/NO signalling pathway.     

Interleukin-8 as a prognostic serum marker in canine mammary gland neoplasias

Mammary gland tumors in female dogs are an excellent model for the clinic-pathological, diagnostic and prognostic investigation of mammary neoplasias. Prognostic and predictive markers are effective in research and routine diagnosis. Interleukins play a fundamental role in cancer, with a particular function in tumor growth, invasion and metastatic potential. Interleukin-8 (IL-8) is known to possess tumorigenic and pro-angiogenic properties, and its overexpression is seen in a number of human tumors. IL-8 serum levels were determined and correlated with the clinic-pathological features and clinical evolution of mammary gland neoplasias in female dogs. IL-8 was measured by an immunoenzymatic assay in 30 female dogs with mammary neoplasias within a 12 month follow-up and in 50 control animals. The correlation between IL-8 concentration and clinical parameters was investigated. A statistically significant difference in the IL-8 serum levels was found in tumor-bearing dogs compared to the controls. In addition, when the individual parameters were evaluated, IL-8 content showed a positive correlation with the tumor progression, lymph node involvement, recurrence and death. Single and multivariate analyses showed associations between tumor recurrence, metastasis, high clinical staging and high IL-8, and also with the death risk. This was also consistent with the high IL-8 content in dogs showing tumor recurrence and metastasis. IL-8 superexpression has been detected in a number of human tumors, usually associated with a poor prognostic. Besides promoting angiogenesis, IL-8 is strongly related with the metastatic phenotype of mammary tumor cells. High IL-8 concentration was found in mammary gland cancer patients with advanced disease stages. Our results show that IL-8 can be used as a non-invasive prognostic marker for mammary gland cancer, and can be useful for the prediction of disease progression and recurrence in dogs with mammary neoplasias. The increased level of this cytokine acts as an independent prognostic marker of survival and the identification of animals with the poor prognostic.     

Diversity and abundance of arbuscular mycorrhizal fungi spores under different coffee production systems and in a tropical montane cloud forest patch in Veracruz, Mexico

We evaluate the arbuscular mycorrhizal fungi (AMF) community as measured by spores in different coffee production systems (at the depth of 0?C15?cm). In addition, we analyze the similarities between the AMF communities in coffee production systems and those that occur in a tropical montane cloud forest patch in order to evaluate the capacity of coffee production systems to preserve the native AMF community. We carried out four samplings in five coffee production systems representative of a vegetation structure gradient, and in a forest. From 120 soil samples, 33 morphospecies were detected. In all the sites, the dominant morphospecies were Glomus clarum and Glomus sp. 3. We found no significant difference in AMF spore richness between sites. Diversity was similar in most of the coffee production systems. Significant differences were only detected in spore abundance; during the dry season the forest, shaded traditional rustic system and shaded simple system presented the highest spore abundance. With the exception of one species exclusive to the forest, the coffee production systems all share the same AMF species as the forest. The coffee production systems with the greatest similarity to cloud forest were the shaded traditional rustic system and the shaded simple system. It is suggested that control of weeds and fertilization could be important factors influencing the composition and abundance of AMF spores in coffee production systems.