Aim of this study was to test the reliability of Trypan blue/Giemsa staining to evaluate sperm membrane integrity, acrosomal intactness and morphology in stallion to verify whether it could be ...applied in vitro as useful tool for sperm fertilizing ability. Fertility data on inseminated mares were collected to evaluate the relationship of sperm quality to pregnancy rates. Forty‐one ejaculates were collected from 3 stallions of Salernitano Horse Breed and evaluated for gross appearance, volume, visual motility and membrane integrity with Trypan blue/Giemsa staining and thirty‐five mares were inseminated during the breeding season from April to July. Differences among stallions were found in volume, sperm concentration (p < 0.05) and visual motility (p < 0.01). A decrease in sperm motility, concentration (p < 0.05) and total sperm number was found in June–July (p < 0.01). Live sperm with intact acrosome (LSIA) and proximal droplets (PD) were lower (p < 0.01) in June–July, while acrosome reacted sperm (ARS) percentage increased (p < 0.05). No fertility differences were found among stallions with an average fertility per cycle of 44.6% and a pregnancy rate of 68.6%. Higher percentages of LSIA were found in the ejaculates used to inseminate mares that became pregnant vs those used in mares not pregnant (p < 0.05). The significance of LSIA as test variable to verify the reliability of Trypan blue/Giemsa staining was confirmed by Receiver operating characteristic ROC analysis and the sensitivity of the test was 85% at a cut‐off value of 48% LSIA. Trypan blue‐Giemsa showed to be an accurate method that can be applied on field to evaluate sperm membrane integrity and to identify poor‐quality ejaculates.
The search for self-sustaining, low-input, diversified, and energy-efficient agricultural systems is now a major concern of many researchers, farmers, and policymakers worldwide. A key strategy in ...sustainable agriculture is to restore functional biodiversity of the agricultural landscape. Biodiversity performs key ecological services and if correctly assembled in time and space can lead to agroecosystems capable of sponsoring their own soil fertility, crop protection and productivity. There is consensus that at least some minimum number of species is essential for ecosystem functioning under constant conditions and that a larger number of species is probably essential for maintaining the stability of ecosystem processes in changing environments. Determining which species have a significant impact on which processes in which ecosystems, however, remains an open empirical question. Keywords: functional biodiversity, sustainability, suppressivity, humification.
We present a novel method for the directed evolution of polypeptides, which combines in vitro compartmentalization and covalent DNA display. A library of linear DNA fragments is co-packaged with an ...in vitro transcription/translation mixture in the compartments of a water-in-oil emulsion. Experimental conditions are adjusted so that, in most cases, one compartment contains one DNA molecule. The DNA fragments encode fusion proteins containing a DNA-methyltransferase (M.Hae III), which can form a covalent bond with a 5-fluorodeoxycytidine base at the extremity of the DNA fragment. The resulting library of DNA–protein fusions is extracted from the emulsion and DNA molecules displaying a protein with desired binding properties are selected from the pool of DNA–protein fusions by affinity panning on target antigens. We applied this methodology in model selection experiments, using specific ligands for the capture of peptides and globular proteins bound to DNA. We observed enrichment factors >1000-fold for selections performed in separate emulsions and up to 150-fold for selections performed using mixtures of DNA molecules. M.Hae III could be fused to small globular proteins (such as calmodulin and fibronectin domains), which are ideally suited for the generation of combinatorial libraries and for the isolation of novel binding specificities.
ABSTRACT
Two‐phase pomace represents an important environmental problem in Mediterranean areas. With its high organic content, direct application of two‐phase pomace in the field is recommended to ...improve soil organic carbon levels and fertility. However, this does not consider any antagonistic effects that this application might have on root proliferation and biomass partitioning. We studied the effects of untreated two‐phase pomace on properties of growth substrate, and on shoot and root growth and biomass allocation of potted olive plantlets. A pot experiment was carried out in a greenhouse over 90 days, with five levels of two‐phase pomace and using two olive cultivars. The effects on shoot growth, leaf pigment content and maximum quantum efficiency of photosystem II (ΦPSII) were assessed each month. After 90 days, the shoot and root biomass of the olive plants was quantified, along with total organic matter, and carbon, nitrogen and polyphenol contents of the growth substrate and shoots, and the fine root nutritional status. Two‐phase pomace increased the total organic matter, total nitrogen and polyphenol contents of the growth substrate. It significantly altered biomass partitioning in the olive plantlets, with reduced shoot dry mass and leaf area, and new shoot formation. It also increased fine and total root dry mass for all two‐phase pomace levels except 40%. There were no significant differences in leaf pigment content and ΦPSII across the treatment levels. Therefore, application of untreated two‐phase pomace at more than 4% induces a severe imbalance in olive plantlet biomass partitioning, and shoot and root growth.
•We investigate olive tree responses to complementary water irrigation in Palestine.•We identify minimum irrigation levels to induce significant increases of production.•35% of total estimated water ...requirement helps to increase fruit production per tree.•Oil production per tree doubled when 35% of total water requirement was applied.•Tested water irrigation regimes did not affect the vegetative growth of the shoots.
Prolonged drought stress in Palestine is one of the major limiting factors in the production and yield of the fruit of the olive tree, as this directly affects crop load, oil production per tree, oil quality and alternate bearing. The objective of the present study was to investigate the use of limited amounts of water as complementary irrigation to improve olive fruit growth and yield at harvest. Field-grown adult olive trees (Olea europaea L. cv. Nabali Baladi) were selected in three villages on the northern West Bank, and they were subjected to different complementary irrigation regimes from June to September, 2010. They were thus supplied with 1, 3, 6m3 water irrigation per tree over this entire period, corresponding to 2.4%, 7.1%, 14.2% of the whole seasonal water requirement per tree (42m3 water), respectively. Additional five rain-fed trees per site were used as controls. In one of the three villages, an additional treatment with 15m3 (35.6% of the total need) water irrigation per tree, was also applied. The results showed that the fruit size, pulp-to-pit ratio (on a fresh and dry weight basis) and 1-year-old mixed shoots were not affected by these water irrigation regimes. Total fruit yield per tree increased as the water irrigation increased, with the greatest effects for the highest water irrigation treatment, due to a higher number of fruits per tree (apparently due to lower fruit abscission). This study demonstrates that complementary water irrigation of olive trees to 35% of the whole seasonal water requirement can produce positive effects on olive fruit production in Palestine.
It is now accepted that the growth of solid tumours is dependent on their capacity to acquire a blood supply, and much effort has been directed towards the development of agents (known as ...anti-angiogenics) that disrupt this process. More recently, it has become apparent that targeted destruction of the established tumour vasculature is another avenue for exciting therapeutic opportunities. In this article, we present evidence that vascular targeting is an effective antitumour strategy in animal models, describe strategies for identifying putative tumour vascular targets and discuss future prospects for vascular targeting in the clinic.
Abstract Background/Aim Prognosis assessment in surgical patients with hepatocellular carcinoma (HCC) remains controversial. The most widely used HCC prognostic tool is the Barcelona Clinic Liver ...Cancer (BCLC) classification, but its prognostic ability in surgical patients has not been yet validated. The aim of this study was to investigate the value of known prognostic systems in 400 Italian HCC patients treated with radical surgical therapies. Methods We analyzed a prospective database collection (400 surgical, 315 nonsurgical patients) observed at a single institution from 2000 and 2007. By using survival times as the only outcome measure (Kaplan–Meier method and Cox regression), the performance of the BCLC classification was compared with that of Okuda, Cancer of the Liver Italian Program, United Network for Organ sharing TNM, and Japan Integrated Staging Score staging systems. Results Two hundred twenty-five patients underwent laparotomy resection; 55, laparoscopic procedures (ablation and/or resection); and 120, liver transplantations. In the surgical group, BCLC proved the best HCC prognostic system. Three-year survival rates of patients in BCLC Stages A, B, and C were 81%, 56%, and 44% respectively, ( P < .01); whereas all other tested staging systems did not show significant stratification ability. When all 715 HCC patients were considered, surgery proved to be a significant survival predictor in each BCLC stage (A, B, and C). Conclusions BCLC staging showed the best interpretation of the survival distribution in a surgical HCC population. The BCLC treatment algorithm should consider the role of surgery also for intermediate-advanced stages of liver disease.
PDF
