Effects of potato plants expressing a barley cystatin on the predatory bug Podisus maculiventris via herbivorous prey feeding on the plant.

The aim of this study was to assess the effects of potato plants expressing a barley cystatin on a potentially cystatin-susceptible natural enemy by predation on susceptible and non-susceptible preys feeding on the plant. We have focussed on the impact of the variant HvCPI-1 C68 --> G, in which the only cysteine residue was changed by a glycine, on the growth and digestive physiology of the Colorado potato beetle (CPB), Leptinotarsa decemlineata, and the Egyptian cotton leafworm (ECW), Spodoptera littoralis. Moreover, we have studied the prey-mediated effects of the barley cystatin at the third trophic level, using the predatory spined soldier bug (SSB), Podisus maculiventris, as a model.

Increasing the amount of n-3 fatty acid in meat from young Holstein bulls through nutrition.

Fifty-four Holstein bulls were blocked by initial BW (301 +/- 7.4 kg) and randomly assigned to 6 treatments following a 3 x 2 factorial arrangement, with 3 concentrate lipid levels (5, 8, and 11% of DM) and 2 lipid sources (whole canola seed and whole linseed), with the objective of evaluating the possibility of increasing the content of n-3 fatty acids in meat. Concentrates (mostly corn meal) were isonitrogenous and isocaloric.

Novel approach to control sulfamethazine misuse in food-producing animals by hair analysis.

The presence of sulfamethazine residues in pig and calf hair was compared with the residual levels encountered in the corresponding edible tissues (liver and muscle) as a consequence of drug administration. Sulfamethazine up to 84.7 mg kg-1 was found in calf hair samples after a pharmacological treatment, with a significant effect of hair pigmentation.

Ternary complex formation between HvMYBS3 and other factors involved in transcriptional control in barley seeds.

The SHAQKYF R1MYB transcription factor (TF) HvMYBS3 from barley is an activator of gene expression both during endosperm development and in aleurone cells upon seed germination. Its mRNA was detected as early as 10 days after flowering in developing barley endosperm, with a peak at 18 days, and in aleurone cells at 8 h after water imbibition, as shown by Northern blot and in situ hybridization analyses. The HvMYBS3 protein expressed in bacteria binds to oligonucleotides containing a GATA core derived from the promoters of: (i) the developing endosperm gene Itr1 (5'-GATAAGATA-3') encoding trypsin inhibitor BTI-CMe, and (ii) the post-germinating aleurone gene Amy6.

Different European-type vaccines against porcine reproductive and respiratory syndrome virus have different immunological properties and confer different protection to pigs.

Immunization of piglets with two different European-type modified live vaccines against porcine reproductive and respiratory syndrome (PRRS) virus produced different outcomes. After vaccination, pigs became viremic (42 days), neutralizing antibodies did not develop, and frequencies of virus-specific gamma-interferon-secreting cells (IFN-gamma-SC) were low. Levels of interleukin-10 (IL-10) produced by peripheral blood mononuclear cells (PBMC) seemed to inversely correlate with interferon-gamma responses.

Response surface methodology for the optimization of keratinase production in culture medium containing feathers produced by Kocuria rosea.

A 43-fold increase in keratinase production by Kocuria rosea was achieved in batch fermentation using response surface methodology. Factorial designs were used to select the components of a culture medium that showed a significant effect on keratinase production. An orthogonal-central composite experimental design was performed, with only two (feathers and magnesium) from nine initial compounds being further analyzed by response surface methodology.

HvMCB1, a R1MYB transcription factor from barley with antagonistic regulatory functions during seed development and germination.

The functional analysis of hydrolase gene promoters induced by gibberellin (GA) in barley aleurone cells upon germination has identified a tripartite GA-response complex (GARC) containing a 5'-TATCCAC-3' box as well as the GA-responsive element (GARE) recognized by GAMYB and the pyrimidine box interacting with the DOF transcription factors BPBF and SAD. We show here that the MCB1 gene encoding a R1MYB protein binds to the 5'-TATCCAC-3' (GATA core) box in vitro and is a transcriptional repressor of a GA-induced amylase (Amy6.4) promoter in bombarded aleurone layers.

Evolution of ORF5 of Spanish porcine reproductive and respiratory syndrome virus strains from 1991 to 2005.

ORF5 sequences of porcine reproductive and respiratory syndrome virus (PRRSV) were analysed to determine genetic diversity, codon usage, positive and negative selection sites and potential changes in the predicted glycoprotein 5 (GP5). A hypothetical GP5 containing all selected sites was constructed to determine its characteristics. These sequences corresponded to isolates obtained 10 years apart (1991-1995, 18 strains) and a second set (n = 46) from 2000 to 2005.

Quality pork genes and meat production.

Functional genomics, including analysis of the transcriptome and proteome, provides new opportunities for understanding the molecular processes in muscle and how these influence its conversion to meat. The Quality Pork Genes project was established to identify genes associated with variation in different aspects of raw material (muscle) quality and to then develop genetic tools that could be utilized to improve this quality. DNA polymorphisms identified in the porcine PRKAG3 and CAST genes illustrate the impact that such tools can have in improving meat quality.

Immune responses of pigs after experimental infection with a European strain of Porcine reproductive and respiratory syndrome virus.

The purpose of this experiment was to study the immune response of pigs during an experimental infection with a European strain of Porcine reproductive and respiratory syndrome virus (PRRSV). Five pigs were challenged intranasally with PRRSV strain VP21 and another five were kept as controls. Clinical course and humoral and cell-mediated responses were monitored for 70 days post-infection (p.

The DOF protein, SAD, interacts with GAMYB in plant nuclei and activates transcription of endosperm-specific genes during barley seed development.

The DOF protein, SAD, previously shown to be a transcriptional activator in barley aleurone cells upon seed germination, also has an important role in gene regulation during endosperm development. mRNA was detected in early (10 days after flowering) developing barley seeds where it accumulated in the starchy endosperm, aleurone cells, nucellar projection, vascular tissues and the immature embryo, as shown by RT-PCR and in situ hybridization analyses. The SAD protein, expressed in bacteria, binds to oligonucleotides containing the prolamine box, 5'-A/TAAAG-3'sequence, derived from the promoter regions of the endosperm-specific genes Hor2 and Itr1, encoding a B-hordein and trypsin-inhibitor BTI-CMe, respectively.

Use of ELISPOT and ELISA to evaluate IFN-gamma, IL-10 and IL-4 responses in conventional pigs.

ELISPOT and ELISA were standardised for pig interferon-gamma (IFN-gamma), interleukin-10 (IL-10) and interleukin-4 (IL-4) with the aim to study the evolution of the immune response in conventional pigs from birth to 6 months of age and also to compare results of both techniques. Five pigs were bled at 1, 6, 9, 12 and 22 weeks of age and peripheral blood mononuclear cells (PBMC) were stimulated with phytohemagglutinin. The frequencies of cytokine secreting cells (CSC) and the levels of secreted cytokines were compared.

The strawberry gene Cyf1 encodes a phytocystatin with antifungal properties.

An EST, encoding a strawberry phytocystatin (PhyCys) obtained from a developing fruit of Fragariaxananassa cv. Elsanta has been characterized. The corresponding gene (Cyf1) had three introns interrupting its ORF that codes for a protein (FaCPI-1) of 235 amino acid residues with a putative signal peptide of 29 residues and an estimated molecular mass for the mature protein of 23.

Comparative phylogenetic analysis of cystatin gene families from arabidopsis, rice and barley.

The plant cystatins or phytocystatins comprise a family of specific inhibitors of cysteine proteinases. Such inhibitors are thought to be involved in the regulation of several endogenous processes and in defence against pests and pathogens. Extensive searches in the complete rice and Arabidopsis genomes and in barley EST collections have allowed us to predict the presence of twelve different cystatin genes in rice, seven in Arabidopsis, and at least seven in barley.

The barley cystatin gene (Icy) is regulated by DOF transcription factors in aleurone cells upon germination.

A genomic clone (Icy gene) encoding a barley cystatin has been characterized. The gene contains one intron interrupting its ORF that encodes a protein of 107 amino acid residues. A DNA fragment of -1058 bp upstream of the ATG translation initiation codon has been sequenced and several promoter deletions fused to the beta-glucuronidase (GUS; uidA gene) reporter gene obtained.

Extended coasting duration exerts a negative impact on IVF cycle outcome due to premature luteinization.

Coasting, or withholding gonadotrophin administration while maintaining gonadotrophin-releasing hormone analogue until oestradiol drops to a safe concentration, is an alternative approach to prevent ovarian hyperstimulation syndrome (OHSS) in high responder patients. However, the length of this procedure has not been precisely studied. This paper is a retrospective study of 132 patients who showed a high response (oestradiol > 4500 pg/ml and/or more than 20 follicles > 17 mm) to ovarian stimulation and were coasted due to their high risk of developing OHSS, and evaluated the impact of the duration of coasting on IVF cycle outcome.

Immunological profile of patients awaiting a renal transplant.

Background: A renal transplant is the best possible treatment for patients with terminal renal failure. Advances in the development of techniques of screening of pre-formed antibodies have contributed to a notable improvement in the results obtained with allogenic transplants.

Methods: The aim of the present work is to study the nature, class, isotype and specificity of antibodies detected in patients awaiting renal transplantation at Complejo Hospitalario Universitario Juan Canalejo, as well as their relation with the level of anti-human leucocyte antigen (HLA) sensitization.

Construction of an immunotoxin with the pore forming protein StI and ior C5, a monoclonal antibody against a colon cancer cell line.

Sticholysin I (StI), a potent cytolysin isolated from the sea anemone Stichodactyla helianthus, was linked to the monoclonal antibody (mAb) ior C5. StI acts by forming hydrophilic pores in the membrane of the attacked cells leading to osmotic lysis. ior C5 is a murine IgG1, which recognizes the tumor associated antigen (TAA) ior C2.

In vivo dose perturbation effects of metallic dental alloys during head and neck irradiation with intensity modulated radiation therapy.

A patient with base of tongue squamous sell carcinoma, with significant CT artifact-inducing metallic alloy, non-removable dental restorations in both the mandible and maxilla was identified. Simultaneous with IMRT treatment, thermoluminescent dosimeters (TLDs) were placed in the oral cavity. After a series of three treatments, the data from the TLDs and software calculations were analyzed.

Detection of QTL affecting fatty acid composition in the pig.

We present a QTL genome scan for fatty acid composition in pigs. An F2 cross between Iberian x Landrace pigs and a regression approach fitting the carcass weight as a covariate for QTL identification was used. Chromosomes (Chrs) 4, 6, 8, 10, and 12 showed highly significant effects.

Splanchnic perfusion during delayed, hypotensive, or aggressive fluid resuscitation from uncontrolled hemorrhage.

The purpose of this study was to determine the effect of three different fluid resuscitation strategies on splanchnic perfusion in a clinically relevant model of uncontrolled hemorrhage after liver trauma. Anesthetized swine were instrumented with a gastric near-infrared spectroscopy probe (GStO2), a jejunal tonometer (PrCO2), a portal vein catheter (SpvO2, lactate), and an ultrasonic blood flow probe on the superior mesenteric artery. The liver was lacerated to produce uncontrolled hemorrhage and a shock state characterized by a 40-60% decrease in cardiac output and a decrease in mean arterial pressure (MAP) to 42 +/- 1 mmHg.

Inhibition of plant-pathogenic fungi by the barley cystatin Hv-CPI (gene Icy) is not associated with its cysteine-proteinase inhibitory properties.

The recombinant barley cystatin Hv-CPI inhibited the growth of three phytopathogenic fungi (Botrytis cinerea, Colletotrichum graminicola, and Plectosphaerella cucumerina) and the saprotrophic fungus Trichoderma viride. Several mutants of barley cystatin were generated by polymerase chain reaction approaches and both their antifungal and their cysteine-proteinase inhibitory properties investigated. Point mutants R38-->G, Q63-->L, and Q63-->P diminished their capacity for inhibiting papain and cathepsin B, retaining their antifungal properties.

[Geriatric assessment in the primary care].

Background: A good medical care program for the elderly requires a thorough assessment of the functional capacity and limitations of the consulting patients.

Aim: To perform a geriatric assessment to elderly subjects consulting in a public outpatient clinic.

Patients And Methods: A cross sectional assessment of subjects over 60 years old, beneficiaries of the public health care system and consulting in 4 outpatient clinics of a poor community of Metropolitan Santiago.

Quantitation and comparison of anti-Gal-alpha-1,3-Gal antibodies in sera of healthy individuals and patients waiting for kidney transplantation.

Highly sensitized patients have poor prospects of being offered an allogeneic donor transplant, therefore, they could become primary candidates for xenogeneic organ transplantation. Hyperacute rejection is initiated by anti-Gal-(alpha)-1,3-Gal antibodies (xenoAb) and this rejection will have to be avoided if xenotransplantation is introduced into clinical practice. Hence, the aim of the study was to determine if sensitized patients against HLA antigens have higher titers of xenoAb than nonsensitized individuals.