Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).     

烟草线条病毒RT-PCR检测方法

根据已报道的烟草线条病毒外壳蛋白基因序列，设计出一对特异性引物，提取大豆病叶的总RNA，反转录成cDNA，PCR扩增出约400bp大小的产物，产物克隆到pGEM-T Easy载体上，转化大肠杆菌DH 5α，抽提的质粒用Not Ⅰ酶切，出现430bp大小的目标插入片段。经序列测定确认了插入片段的大小为404bp。     

Egg quality determination based on the shape of the lipid vesicle in common dentex, Dentex dentex

The shape of the lipid vesicle (the ratio between the maximal and the minimal diameter and the shape coefficient) of the eggs of Diplodus puntazzo and Sparus aurata is correlated with larval survival rate (%) and can be used as an egg quality marker as reported previously by Lahnsteiner and Patarnello (2005) . In the present study, this method for egg quality determination has also been validated for common dentex, Dentex dentex, after introducing several modifications in the calculation and evaluation procedure in order to obtain higher sensitivity. In the present study, it could be demonstrated that the shape of the lipid vesicle is not only correlated with the hatching rate but also with the survival rate of 1, 2 and 3 days post hatch larvae and therefore it can also be used as an indicator of larval viability at early stages of development.     

Maturational development of drug-metabolizing enzymes in dogs

A qualitative and quantitative assessment was made of the development of hepatic drug-metabolizing enzymes (DME) in dogs as part of a study of the ability of animal test species to metabolize drugs. The following DME variables were measured in this study: amount of cytochromes P-450 and b5; activity of the NADPH and NADH-dependent reductases associated with each of these cytochromes; activity of cytochrome P-450-mediated enzymes, including aniline and coumarin hydroxylases, aminopyrine N-demethylase, and 7-ethoxycoumarin O-deethylase; activity of a uridine diphosphoglucuronic acid glucuronyl transferase with 4-methylumbelliferone as substrate; and glutathione-S-transferase activities, with dinitrochlorobenzene and dichloronitrobenzene as substrates. Most enzyme components had achieved maximal amount or activity by the fifth to eighth week after birth; they tended to decrease after weaning, although the activity of dichloronitrobenzene-glutathione transferase in geriatric dogs (312 to 525 weeks old) was approximately twofold greater than that of 8-week-old pups. There were no gender-related differences in any of the enzyme amounts or activities determined. Individual variation was pronounced even in the homogeneous colony from which these dogs were obtained.     

China's role in the global forest sector: how will the US recovery and a diminished Chinese demand influence global wood markets?

Over the last decade, while the size of China's economy more than doubled, China has simultaneously become a major producer and exporter of forest products. Although China's domestic supply of wood is significantly constrained both by a limited natural supply and by conservation-oriented policies, the country is increasingly regarded as the world's “wood workshop.” Furthermore, China is the largest driver of demand for the trade in tropical logs and is becoming a significant driver of demand for trade in coniferous logs. In this paper, we describe a spatial equilibrium model adapted to study forest sector markets and policies that affect them. We present the model and the result of two alternative future scenarios. The first scenario analyzes the impact on global forest products markets of a US recovery in wood markets. The second scenario examines the effect on global forest products markets of decelerating growth in Chinese demand for wood products. Through these two scenarios, the modeling output sheds light on the role China's wood products markets have on resource supply and trade around the world. The trade model shows substantial potential changes in global prices, production, and trade activity associated with the recovery in domestic demand in the USA.     

A Large‐scale,Rapid Public Health Response to Rabies in an Organ Recipient and the Previously Undiagnosed Organ Donor

This article describes and contrasts the public health response to two human rabies cases: one organ recipient diagnosed within days of symptom onset and the transplant donor who was diagnosed 18 months post‐symptom onset. In response to an organ‐transplant‐related rabies case diagnosed in 2013, organ donor and recipient investigations were conducted by multiple public health agencies. Persons with potential exposure to infectious patient materials were assessed for rabies virus exposure. An exposure investigation was conducted to determine the source of the organ donor's infection. Over 100 persons from more than 20 agencies spent over 2700 h conducting contact investigations in healthcare, military and community settings. The 564 persons assessed include 417 healthcare workers [5.8% recommended for post‐exposure prophylaxis (PEP)], 96 community contacts (15.6% recommended for PEP), 30 autopsy personnel (50% recommended for PEP), and 21 other persons (4.8% recommended for PEP). Donor contacts represented 188 assessed with 20.2% recommended for PEP, compared with 5.6% of 306 recipient contacts recommended for PEP. Human rabies cases result in substantial use of public health and medical resources, especially when diagnosis is delayed. Although rare, clinicians should consider rabies in cases of encephalitis of unexplained aetiology, particularly for cases that may result in organ donation.     

Reduced-calorie Avocado Paste Attenuates Metabolic Factors Associated with a Hypercholesterolemic-high Fructose Diet in Rats

The objective of this study was to evaluate the effect of reduced-calorie avocado paste on lipid serum profile, insulin sensitivity, and hepatic steatosis in rats fed a hypercholesterolemic-high fructose diet. Thirty five male Wistar rats were randomly separated in five groups: Control group (ground commercial diet); hypercholesterolemic diet plus 60 % fructose solution (HHF group); hypercholesterolemic diet plus 60 % fructose solution supplemented with avocado pulp (HHF+A group); hypercholesterolemic diet plus 60 % fructose solution supplemented with reduced-calorie avocado paste (HHF+P group); and hypercholesterolemic diet plus 60 % fructose solution supplemented with a reduced-calorie avocado paste plus fiber (HHF+FP group). The A, P, and FP were supplemented at 2 g/kg/d. The study was carried out for seven weeks. Rats belonging to the HHF group exhibited significantly (P?≤?0.05) higher total cholesterol, triglycerides, and insulin levels in serum as well as lower insulin sensitivity than the control group. Supplementation with reduced-calorie avocado paste showed a significant (P?≤?0.05) decrease in total cholesterol (43.1 %), low-density lipoprotein (45.4 %), and triglycerides (32.8 %) in plasma as well as elevated insulin sensitivity compared to the HHF group. Additionally, the liver enzymes alanine aminotransferase and aspartate aminotransferase decreased significantly in the HHF-P group (39.8 and 35.1 %, respectively). These results are likely due to biocompounds present in the reduced-calorie avocado paste, such as polyphenols, carotenoids, chlorophylls, and dietary fibre, which are capable of reducing oxidative stress. Therefore, reduced-calorie avocado paste attenuates the effects of a hypercholesterolemic-high fructose diet in rats.     

Organic Fertilization in Traditional Mediterranean Grapevine Orchards Mediates Changes in Soil Microbial Community Structure and Enhances Soil Fertility

Soil microbial populations and their functions related to nutrient cycling contribute substantially to the regulation of soil fertility and the sustainability of agroecosystems. A field experiment was performed to assess the medium‐term effect of a mineral fertilizer and two organic fertilization systems with different nitrogen sources on the soil microbial community biomass, structure, and composition (phospholipid fatty acids, pattern, and abundance), microbial activity (basal respiration, dehydrogenase, protease, urease, β‐glucosidase, and total amount of phosphomonoesterase activities), and physical (aggregate stability) and chemical (total organic C, total N, available P and water‐soluble carbohydrates) properties in a vineyard under semiarid Mediterranean conditions after a period of 10 years. The three fertilization systems assayed were as follows: inorganic fertilization, addition of grapevine pruning with sheep manure (OPM), and addition of grapevine pruning with a legume cover crop (OPL). Both treatments, OPM and OPL, produced higher contents of total organic carbon, total N, available P, water‐soluble carbohydrates, and stable aggregates. The organic fertilization systems increased microbial biomass, shifted the structure and composition of the soil microbial community, and stimulated microbial activity, when compared with inorganic fertilization. The abundances of fungi and G+ bacteria were increased by treatments OPM and OPL, without significant differences between them. Organic and inorganic fertilization produced similar grapevine yields. The ability of the organic fertilization systems for promoting the sustainability and soil biological and chemical fertility of an agroecosystem under semiarid conditions was dependent of the organic N source. Copyright © 2016 John Wiley & Sons, Ltd.     

Potential of a Fusarium eumartii culture filtrate on the screening for wilting resistance in potato

Summary Fusarium solani f.sp. eumartii Carp. Snyder and Hansen (Fusarium eumartii) is a soil inhabitant that induces the so-called Potato Wilt and Stem End Rot disease. Prior to wilting, the pathogen induces peculiar small bronze spots on the leaflets. Failure to isolate F. eumartii from infected leaflets suggests the involvement of a toxin in the disease. The fungus was grown in liquid Richard's medium and thereafter a filtrate was obtained dialyzing (MW cutoff 12,000–14,000) and sterilizing the culture by filtration (0.22 m). Potato leaves treated with both the pathogen or the filtrate showed symptoms of bronze spots and significantly higher electrolyte leakage when compared to controls. Tomato leaves showed neither bronze spots nor electrolyte leakage after plant inoculation with the pathogen or with the filtrate treatment. Both, the absence of visible symptoms and the lack of electrolyte leakage in tomato could be associated to a certain degree of host specificity of the F. eumartii filtrate towards potato. The filtrate also induced symptoms similar to infections by F. eumartii in adult plants and in vitro plantlets of cultivars Huinkul MAG and Kennebec. Callus responses to the filtrate were related to responses of the cultivars to the pathogen in greenhouse. These results show the potential of the culture filtrate of F. eumartii for use in screening for wilting resistance.