Identification of olive (Olea europaea) pulp proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nano-liquid chromatography tandem mass spectrometry

Proteins in the pulp of olive ( Olea europaea ) constitute a minor fraction. They have been sparsely studied despite their suggested role in oil stability and olive allergenicity. The analysis of a pulp protein extract by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a major band at 24 kDa that was subjected to tryptic in-gel digestion. Peptide extracts were analyzed by MALDI-TOF MS and nanoLC-MS/MS. The use of different search engines enabled the assignment of a number of fragmentation spectra to peptide sequences, identifying a major band as a thaumatin-like protein and other low-abundant proteins such a drought-induced protein SDi-6-like, an acyl carrier protein, Cu/Zn and Mn superoxide dismutases, a small heat shock protein, and an ATP-dependent protease subunit. Many of the produced spectra did not give good matches in the database searches, due to the scarce presence of O. europaea entries in protein databases. Nevertheless, a huge number of spectra corresponded to peptides, which showed a high degree of homology with others from sequenced organisms. These results proved that database searching with MS/MS spectra constitutes a promising approach for the characterization of olive pulp proteins.     

Pathological effects of Cryptostegia venusta toxicity in goats and rats

Here, we aimed to determine the toxicity of Cryptostegia venusta in goats and rats. We orally administered a single 60 g dose of shredded C. venusta leaves per kilogram of body weight to three goats. The animals were necropsied after death, and tissue sections were collected and routinely processed for histopathological analyses. Additionally, we separated 25 adult male Wistar rats (each weighing about 150 g) into five groups: an untreated control group and groups orally treated with 1, 3, 10, or 60 g/kg doses. Rats were sacrificed 72 h after administration of the C. venusta extract, and tissue sections collected for histopathological analyses. All goats presented signs of apathy, salivation, frequent urination, and eventually fatigue 4–6 h after receiving C. venusta. Two goats died 20 h after administration, and the third was sacrificed in extremis. The only histopathological finding observed in the goats was lung edema. No rats died during the experimental period or presented any clinical signs or macroscopic lesions. However, both goats and rats exhibited degeneration and multifocal necrosis of cardiac muscle fibers. From our results, we conclude that the C. venusta plant is capable of promoting cardiotoxicity.     

Genetic definition and sequence analysis of Arabidopsis centromeres

High-precision genetic mapping was used to define the regions that contain centromere functions on each natural chromosome in Arabidopsis thaliana. These regions exhibited dramatic recombinational repression and contained complex DNA surrounding large arrays of 180-base pair repeats. Unexpectedly, the DNA within the centromeres was not merely structural but also encoded several expressed genes. The regions flanking the centromeres were densely populated by repetitive elements yet experienced normal levels of recombination. The genetically defined centromeres were well conserved among Arabidopsis ecotypes but displayed limited sequence homology between different chromosomes, excluding repetitive DNA. This investigation provides a platform for dissecting the role of individual sequences in centromeres in higher eukaryotes.     

The chromosomal location of the embryo plus scutellum alcohol dehydrogenase isozymes in the hexaploid wheat kernel

Summary Analysis of individual parts of Triticum aestivum L. and T. turgidum var. durum kernels showed two classes of alcohol dehydrogenase patterns: 1) A three banded pattern (ADH-1, ADH-2 and ADH-3) for endosperm (Ed) and 2) a seven banded pattern (ADH-F1, ADH-F2, ADH-1, ADH-2, ADH-3, ADH-S1 and ADH-S2) for embryo (E), scutellum (S) and embryo plus scutellum (E+S). The use of nullitetrasomic and ditelosomic series of Chinese Spring wheat allows alcohol dehydrogenase to be associated with arms 4A, 4BL and 4DS.A five subunit forming dimer hypothesis is postulated to explain the newly reported seven isozyme pattern. Subunits , and are coded by chromosome arms 4A, 4BL and 4DS respectively, and the newly postulated and are coded also by chromosome arm 4BL.     

Comparison of a gas detection tubes test with the traditional alkaline trap method to evaluate compost stability

Compost stability is an important parameter of compost quality. Among tests proposed to evaluate compost stability, microbial respiration is one of the better accepted tests. Variations in rates of CO₂ evolution during composting were studied in two pilot pruning waste piles using a windrow composting system. To measure the CO₂ production rate, two methods were compared: the alkaline trap test and gas detection tubes. Both respiration tests indicated increasing compost stability with processing time, but CO₂ evolution rates from the alkaline trap method were higher than values from the gas detection tube method. A first-order kinetic equation was used to describe CO₂ evolution over time. A linear relationship (r=0.81, p<0.01) was found between the two methods. Although both methods could distinguish unstable compost from stable compost, CO₂ detection tubes were easier to use and gave results in a shorter period of time.     

Effect of protease EPg222 obtained from Penicillium chrysogenum isolated from dry-cured ham in pieces of pork loins

The fungal protease EPg222 obtained from Penicillium chrysogenum Pg222 isolated from dry-cured ham, was assayed for proteolytic activity in a meat model system based on sterile pieces of pork loins for 32 days. Treated samples showed a significative reduction of total high ionic strength-soluble proteins during the incubation period, as compared with a control incubated without enzyme, both on the surface and in the depth. SDS-PAGE analysis of this protein fraction showed higher hydrolysis of the main myofibrillar proteins H-meromyosin, actin, and tropomyosin in treated samples. Non-protein and amino acidic nitrogen were detected in higher amounts in enzyme-added than in control pieces of loins, both on the surface and in the depth. Thus, addition of enzyme EPg222 to whole pieces of meat results in an increase of protein hydrolysis. The effect of this enzyme could be of great interest for stimulating proteolysis in whole dry-cured meat pieces.     

Use of three fodder trees in the feeding of goats in the subhumid tropics in Mexico

Chemical composition, in vitro gas production with and without polyethyleneglycol (PEG-4000 MW), and in vitro digestibility of dry matter (IVDMD) and organic (IVOMD) foliage from Pithecellobium dulce, Gliricidia sepium and Haematoxylum brasiletto were determined. The preference test was run for 15 days: the first 10 days as adaption period and the 5 days served as evaluation period. It was conducted in ten developing female Creole goats of 6 months old, weighing 14?±?2.0 kg in order to determine goat preference for any of the three foliages. Productive performance of 35 male creole kids of 6 months old (14?±?3.0 kg) was also determined by ad libitum feeding of the foliage of the tree: the 30 and 15 % of each of the P. dulce (T1, T2), G. sepium (T3, T4), and H. brasiletto (T5, T6) foliages were added to the experiment diets, while T7 served as control diet that did not contain any foliage. The crude protein (CP), total phenols (TP), condensed tannins (CT), IVDMD, and IVOMD were different among the foliages. The PEG determined the biological activity of the TP and CT of H. brasiletto. Goats preferred to consume the foliage of P. dulce because of its higher content of CP and IVDMD and low content of TP and CT. In the productive response, dry matter intake (DMI) was higher in kids fed T1 diet and was stimulated by higher IVDMD and IVOMD, which resulted in the higher daily weight gain (DWG). The contribution with TP and CT of H. brasiletto to T5 and T6 and the rejection by the animals of G. sepium in T3 and T4 explain the negative effects on the DMI and the DWG. Findings of the study suggested higher kid performance for P. dulce foliage. Possible attributes may include its better CP, low TP and CT, and higher digestibility.     

Urea treated maize straw for small-scale dairy systems in the highlands of Central Mexico

Maize straw is the main roughage for dairy herds in campesino farms in central Mexico. The objective was to evaluate feeding milking cows on maize straw treated with 40 g/kg DM of urea (A) or untreated straw (B), and 3.0 kg/d of 18% CP concentrate. Twenty-four Holsteins in late lactation from 8 farmers were sorted in two groups: sequence A-B-A or B-A-B; periods were 28 days. Mean daily milk yield for the last two weeks per period, and live-weight and body-condition score every 14 days were used for analysis. Maize straw was ad libitum. Chemical analysis and in vitro digestibility were analysed by Student’s t test, animal variables by a switch-back design. ‘A’ had 44.3 g/kg DM more CP and 106.5 g/kg DM higher in vitro digestibility than ‘B’ (710 g/kg DM ± 0.75 ‘A’ vs. 603.5 g/kg DM ± 1.44 ‘B’). Despite higher digestibility and intake, there were no differences (P > 0.05) for milk yield, live-weight or body-condition score, although there were in straw intake (P < 0.05). Cows on ‘A’ ate 1.7 kg/cow/day more straw DM than on ‘B’. Lack of response did not offset higher feeding costs although margins were high. Lack of response is attributed to short length of periods and late lactation of cows.     

Holocene alluvial morphopedosedimentary record and environmental changes in the Bardenas Reales Natural Park (NE Spain)

A Holocene alluvial morphostratigraphy, based on geomorphological mapping, sedimentological analysis, soil development and radiocarbon chronology is presented for the semiarid Blanca basin in the Bardenas Reales Natural Park (NE, Spain). Four morphopedosedimentary alluvial units (MU1, MU2, MU3 and MU4) were differentiated, which may correspond to two palaeogeographical stages. The first one includes the morphopedosedimentary unit MU1 and is the most extensive in the area. It is made up of a complex aggradation (fill up) sequence comprising a braided channel system, a sand–mud flat (9200–7900 cal BP) and a playa-lake environment related to a nearly closed depression. An intensive fluvial entrenchment period is recognized, leading the opening of the Blanca basin, before the second stage that includes the units MU2, MU3 and MU4. This set of units forms a sequence of cut-and-fill and nested fill terraces recording the alluvial activity in the Blanca basin during the last 6 ka. The MU2 unit, 5900–3300 cal BP in age, represents a period of high geomorphic instability with rapid alluvial sedimentation/channel incision oscillating changes. On the other hand, the MU3 unit has an age of 1050–500 cal BP and appears as a thin extensive alluvial cover overlying previous units. Finally, the MU4 unit comprises superposition of numerous flood depositional sequences, centimetric in scale, dated from 450–10 cal BP. The established soil chronosequence supports the numerical chronology of the four units. These accumulation periods (MU1, MU2, MU3, MU4) are associated with dry phases and/or high hydrological variability (floods and droughts) at a regional scale. Changes on the North Atlantic atmospheric circulation mode over Southern Europe may provide the key to explain the nature of alluvial activity during Holocene in Bardenas Reales.