Evaluation of the genetic diversity of microsatellite markers among four strains of Oreochromis niloticus.

Different strains of Nile tilapia can be found worldwide. To successfully use them in breeding programs, they must be genetically characterized. In this study, four strains of Nile tilapia - UFLA, GIFT, Chitralada and Red-Stirling - were genetically characterized using 10 noncoding microsatellite loci and two microsatellites located in the promoter and first intron of the growth hormone gene (GH). The two microsatellites in the GH gene were identified at positions -693 to -679 in the promoter [motif (ATTCT)8 ] and in intron 1 at positions +140 to +168 [motif (CTGT)7 ]. Genetic diversity was measured as mean numbers of alleles and expected heterozygosity, which were 4 and 0.60 (GIFT), 3.5 and 0.71 (UFLA), 4.5 and 0.57 (Chitralada) and 2.5 and 0.42 (Red-Stirling) respectively. Genetic differentiation was estimated both separately and in combination for noncoding and GH microsatellites markers using Jost's DEST index. The UFLA and GIFT strains were the least genetically divergent (DEST  = 0.10), and Chitralada and Red-Stirling were the most (DEST  = 0.58). The UFLA strain was genetically characterized for the first time and, because of its unique origin and genetic distinctness, may prove to be an important resource for genetic improvement of Nile tilapia. This study shows that polymorphisms found in coding gene regions might be useful for assessing genetic differentiation among strains.

Growth hormone receptors in the atherinid Odontesthes bonariensis: characterization and expression profile after fasting-refeeding and growth hormone administration.

In order to improve the understanding of pejerrey Odontesthes bonariensis, growth hormone (Gh)-insulin-like growth factor-1(Igf1) axis, O. bonariensis growth hormone receptor type 1 (ghr1) and type 2 (ghr2) mRNA sequences were obtained. Both transcripts were ubiquitously expressed except in kidney, encephalon and anterior intestine. Alternative transcripts of both receptors were found in muscle. Interestingly, two different ghr2 transcripts with alternative polyadenylation (APA) sites located in the long 3' untranslated region (UTR-APA) were also found in liver. Hepatic ghr1, ghr2 and insulin-like growth factor type 1 (igf1) transcript levels were examined under two different metabolic conditions. In the first experimental condition, fish were fasted for 2 weeks and then re-fed for another 2 weeks. Despite igf1 mRNA relative expression did not show significant differences under the experimental period of time examined, both ghr transcripts decreased their expression levels after the fasting period and returned to their control levels after re-feeding. In the second treatment, recombinant O. bonariensis growth hormone (r-pjGh) was orally administered once a week. After 4 weeks of treatment, liver igf1, ghr1 and ghr2 mRNA relative expression increased (13, 4·5 and 2·1 fold, P < 0·05) compared to control values. These results add novel information to the growth hormone-insulin-like growth factor system in teleosts.

Effects of alcohol and polyphenols from beer on atherosclerotic biomarkers in high cardiovascular risk men: a randomized feeding trial.

BACKGROUND AND AIMS: Moderate alcohol consumption exerts a cardioprotective effect, but no studies have evaluated the alcohol-independent cardiovascular effects of the non-alcoholic components of beer. We aimed to evaluate the effects of ethanol and the phenolic compounds of beer on classical and novel cardiovascular risk factors. METHODS AND RESULTS: Thirty-three high risk male volunteers were included in a randomized, crossover feeding trial. After a washout period, all subjects received beer (30 g alcohol/d, 660 mL), the equivalent amount of polyphenols as non-alcoholic beer (990 mL), and gin (30 g alcohol/d, 100 mL) for 4 weeks. All outcomes were evaluated before and after each intervention period. Moderate alcohol consumption increased serum HDL-cholesterol (∼5%), ApoA-I (∼6%), ApoA-II (∼7%) and adiponectin (∼7%), and decreased serum fibrinogen (∼8%), and interleukin (IL)-5 (∼14%) concentrations, whereas the non-alcoholic fraction of beer (mainly polyphenols) increased the receptor antagonist of IL-1 (∼24%), and decreased lymphocyte expression of lymphocyte function-associated antigen-1 (∼11%), lymphocyte and monocyte expression of Sialil-Lewis X (∼16%) and monocyte expression of CCR2 (∼31%), and tumor necrosis factor (TNF)-ß (∼14%) and IL-15 (∼22%) plasma concentrations. No changes were observed in glucose metabolism parameters or in body weight and adiposity parameters. CONCLUSION: The phenolic content of beer reduces leukocyte adhesion molecules and inflammatory biomarkers, whereas alcohol mainly improves the lipid profile and reduces some plasma inflammatory biomarkers related to atherosclerosis.

Isolation and characterization of 20 polymorphic microsatellite loci in the migratory freshwater fish Leporinus obtusidens (Characiformes: Anostomidae) using 454 shotgun pyrosequencing.

Twenty polymorphic microsatellite markers were developed for the Neotropical fish Leporinus obtusidens using a next generation sequencing approach and tested in two other characifomes species, Schizodon platae and Prochilodus lineatus. Microsatellite loci alleles in L. obtusidens ranged between 2 and 20 alleles per locus (mean = 5·7), with expected heterozygosity values ranging from 0·097 to 0·956 (mean = 0·578) and observed heterozygosity values ranging from 0·000 to 0·800 (mean = 0·400) in a sample of 20 specimens from the lower Paraná River (Argentina). Most of these markers will be a valuable tool for captive breeding and stocking programmes, as well as for analyses of population connectivity and genetic structure in this broadly distributed Neotropical migratory fish.

Gazpacho consumption is associated with lower blood pressure and reduced hypertension in a high cardiovascular risk cohort. Cross-sectional study of the PREDIMED trial.

BACKGROUND AND AIM: Hypertension is a major public health problem and a leading cause of death and disability in both developed and developing countries, affecting one-quarter of the world's adult population. Our aim was to evaluate whether the consumption of gazpacho, a Mediterranean vegetable-based cold soup rich in phytochemicals, is associated with lower blood pressure (BP) and/or reduced prevalence of hypertension in individuals at high cardiovascular risk. METHODS AND RESULTS: We selected 3995 individuals (58% women, mean age 67 y) at high cardiovascular risk (81% hypertensive) recruited into the PREDIMED study. BP, weight, and dietary and physical activity data were collected. In multivariate linear regression analyses, after adjustment, moderate and high gazpacho consumption categories were associated with reduced mean systolic BP of -1.9 mm Hg [95% confidence interval (CI): -3.4; -0.6] and -2.6 mm Hg (CI: -4.2; -1.0), respectively, and reduced diastolic BP of -1.5 mm Hg (CI: -2.3; -0.6) and -1.9 mm Hg (CI: -2.8; -1.1). By multiple-adjusted logistic regression analysis, gazpacho consumption was associated with a lower prevalence of hypertension, with OR = 0.85 (CI: 0.73; 0.99) for each 250 g/week increase and OR = 0.73 (CI: 0.55; 0.98) for high gazpacho consumption groups compared to the no-consumption group. CONCLUSIONS: Gazpacho consumption was inversely associated with systolic and diastolic BP and prevalence of hypertension in a cross-sectional Mediterranean population at high cardiovascular risk. The association between gazpacho intake and reduction of BP is probably due to synergy among several bioactive compounds present in the vegetable ingredients used to make the recipe.

[Breast cancer screening in high risk populations]. / Abordaje en el cribado de cáncer mamario en grupos poblacionales de riesgo elevado.

We aim to define which patients make up the populations with high and intermediate risk of developing breast cancer, to review the studies of screening with magnetic resonance imaging in addition to mammography in high risk patients (describing the imaging characteristics of the cancers in this group), to review the studies of screening with magnetic resonance imaging in patients with intermediate risk, and to update the guidelines for screening in patients with high or intermediate risk (based on the recent recommendations of the main scientific societies/American and European guidelines).

Ibero-American consensus on learning outcomes for the acquisition of competencies by medical students through clinical simulation.

OBJECTIVE: Our aim was to reach expert consensus on specific learning outcomes (LOs) that can be achieved through clinical simulation aimed at developing the competencies that medical students need to be able to successfully manage patients and assume general clinical responsibilities. MATERIALS AND METHODS: The six-member scientific committee peer-reviewed Spanish reference documentation (in line with the Bologna Process) on required competencies in medical undergraduate students to select an initial set of 16 competencies that could feasibly be developed through simulation and a corresponding set of 75 LOs. Snowball sampling was used to identify candidates for an international panel of simulation experts. Applying a set of pre-defined criteria, 19 panelists from seven Spanish-speaking regions were recruited to participate in a modified two-round Delphi procedure based on electronic questionnaires and aimed at reaching formal consensus on appropriate LOs for simulated medical training. RESULTS: Final agreement between the panelists was high: no mean score fell below 7.26 of a maximum of 9, and all 75 LOs were agreed on, 74 in the first round and only one requiring the second round. The 16 LOs with mean scores in the top 25th percentile were selected as a set of core LOs to attain via simulation. CONCLUSIONS: This Ibero-American consensus on observable and measurable LOs, reflecting competencies that can feasibly be developed via clinical simulation, is a framework that aims at helping medical schools' plans and delivering specific kinds of undergraduate medical training through simulation. It is also proposed in a set of core LOs as a starting point for less experienced schools to design a simulated training program.

Dietary Polyphenols in the Prevention of Stroke.

Polyphenols have an important protective role against a number of diseases, such as atherosclerosis, brain dysfunction, stroke, cardiovascular diseases, and cancer. Cardiovascular diseases are the number one cause of death worldwide: more people die annually from cardiovascular diseases than from any other cause. The most important behavioural risk factors of heart disease and stroke are unhealthy diet, physical inactivity, tobacco use, and excess alcohol intake. The dietary consumption of polyphenols has shown to be inversely associated with morbidity and mortality by cardio- and cerebrovascular diseases. It is well-known that the protective effects of polyphenols in vivo depend on the grade how they are extracted from food and on their intestinal absorption, metabolism, and biological action with target tissues. The aim of this review was to summarise the relation between polyphenols of different plant sources and stroke in human intervention studies, animal models, and in vitro studies.

[Exercise-induced anaphylaxis syndrome]. / Síndrome de anafilaxia inducida por ejercicio.

In the last two decades a growing incidence of a peculiar form of anaphylaxis that only occurs while carrying out physical exercise has been observed. Within the exercise-induced anaphylaxis syndrome two well differentiated clinical forms are included: systemic cholinergic urticaria and exercise-induced anaphylaxis in the strict sense which can be shown by a classic form or a variant form, more uncommon and with manifestations similar to cholinergic urticaria. Postprandial or food-dependent exercise-induced anaphylaxis is a frequently identified subtype of these last cases. It can be due to an asymptomatic food allergy manifested through physical effort, although in many occasions it is not possible to find a responsible allergen. The diagnosis is settled on the clinical history and specific tests with food allergens. It can be necessary to perform an exercise challenge test with and without previous ingestion. The treatment is preventive and it is based on avoiding the food or the food allergen some hours before the exercise. When it does not depend on foods it is used a prophylactic pharmacotherapy with antihistamines, cromones or sodium bicarbonate. The patient should be well educated on the use of epinephrine in the event of new reactions.

Sonographic findings of additional malignant lesions in breast carcinoma seen by second look ultrasound.

OBJECTIVE: The aim is to show ultrasound (US) findings of additional malignant lesions of breast carcinoma visualized on targeted second-look US that were not identified by mammography or US prior to the time of diagnosis. MATERIALS AND METHODS: A double-blind retrospective review of the US results from January 2008 through August 2010 of 228 patients with known breast cancer was conducted by two expert radiologists. The focus of the review was on the second-look US characteristics (following BI-RADS criteria) of 26 documented additional malignant lesions of the 76 with successful sonographic correlation from the 123 lesions detected by magnetic resonance imaging (MRI). All of them, before the MRI, had an initial mammography and a US with a histopathological biopsy of the primary lesion. RESULTS: Approximately 60 to 70% of the findings were classified as BI-RADS 2 and BI-RADS 3, while assessing the final US category. The review of the second-look US showed the size of the second malignant additional lesion ranged from 3 to 22 mm, of which 90% were smaller than 10 mm and 66% were smaller than 7 mm. CONCLUSIONS: Most additional malignant lesions, nonpalpable carcinomas, which were previously not detected by mammography and US at first-look diagnosis, were detected by a targeted second-look US examination. These lesions were of category BI-RADS 2 and BI-RADS 3 and smaller than 7 mm.

[Serous oligocystic adenoma of the pancreas]. / Adenoma seroso oligoquístico del páncreas.

Serous oligocystic or macrocystic adenoma of the pancreas is a very uncommon morphological variant of what was classically termed microcystic adenoma of the pancreas. This is a benign neoplasm; however, its radiological appearance mimicks that of potentially malignant mucinous neoplasms of the pancreas. Therefore, radiologists need to be familiar with this entity to ensure the most appropriate therapeutic management and help to avoid unnecessary surgery.

Molecular cloning, expression and immunological characterization of pejerrey (Odontesthes bonariensis) growth hormone.

Growth hormone is an essential polypeptide required for normal growth and development of vertebrates. The pejerrey fish, Odontesthes bonariensis, is a South American atherinid freshwater fish considered as a promising species for aquaculture. Although growth hormone has been characterized in a number of fish, there are no published data on the structure of this hormone in atherinids, except that of a related species Odontesthes argentinensis. In this paper, the molecular cloning, expression and immunological characterization of pejerrey growth hormone (pjGH) is described. The predicted amino acid sequence of pjGH cDNA consisted of 204 amino acid residues with an estimated molecular mass of 23 kDa. Amino acid sequence was highly conserved among the two Atheriniformes where the growth hormone sequences are known (99% aa identity), highly to moderately conserve (75-92% aa identity) when compared to the other members of Acantopterigii superorder and clearly less conserved (49-66% identity) when compared to Salmoniformes (Protacanthopterygii), Cypriniformes and Siluriformes (Ostariophysi). A phylogenetic tree depicting the relationship of various teleost GH nucleotide sequences was inferred. Pejerrey GH was produced using recombinant DNA technology in a bacterial system, representing the first time an atherinid growth hormone protein was expressed as a recombinant protein in Escherichia coli. A specific antiserum of this hormone was raised in rabbits and its specificity tested by using Western blot and immunocytochemistry. The distribution of pjGH mRNA was also studied by RT-PCR and Southern blot analysis. The transcript was detected not only in the pituitary gland but also in the testis.

[Treatment with nebulized lidocaine in steroid-dependent asthma]. / Tratamiento con lidocaína nebulizada del asma corticodependiente.

BACKGROUND: The management of glucocorticoid dependent and resistant asthma encompasses a group of problems, affecting as much control of clinical symptoms as the onset of adverse effects to prolonged use of steroids. MATERIALS AND METHODS: Woman 52 years old, steroid dependent asthma fifteen years ago. She has important adverse effects in consequence of the use of systemic glucocorticoids. The employment of methotrexate attained decrease in the dosage of steroids temporarily. The worsening of the clinical situation and deterioration of secondary effects by use of steroids necessitated treatment with nebulized lidocaine. A 2% solution to initial dosage was administered three times daily. The maximal dosage needed was 80 mg three times daily. RESULTS: After three months of lidocaine treatment, the patient was able to reduce oral steroids eventually discontinuing these drugs, and the effects secondary to exogenous hypercortisolism disappeared. Moreover, an improvement in clinical symptoms and quality of life was observed with stability in the pulmonary function. CONCLUSIONS: The use of nebulized lidocaine could be useful and provide alternatives in patients with severe asthma.

Diffusible highly glycosylated protein from Bufo arenarum egg-jelly coat: biological activity.

L-HGP is a highly glycosylated protein from Bufo arenarum egg-jelly coat that diffuses into the surrounding medium when the strings of oocytes are incubated in saline solutions. L-HGP was purified from egg water and the estimated percentage of L-HGP/total protein in egg water was estimated in 30%. In the present study we examine, by indirect immunofluorescence, the effect of L-HGP on acrosome status of homologous spermatozoa. A high percentage (77%) of sperm lost the acrosome when incubated in 10% Ringer solution buffered with 10 mM Tris-HCl, pH 7.6, during 60 min, a condition that resembles egg-jelly osmolarity. The addition of purified L-HGP to the incubation medium prevents acrosome breakdown. The acrosome integrity is maintained for at least 1 hr. This effect is specific for L-HGP at concentration ranging from 0.01 to 0.1 mg/ml since neither BSA nor fetuin seems to have similar activity at similar concentrations. The same effect was observed when spermatozoa were incubated in egg water. Preliminary results suggest that L-HGP binds to B. arenarum spermatozoan membranes.

Bufo arenarum egg jelly coat: purification and characterization of two highly glycosylated proteins.

Egg jelly coats from Bufo arenarum are formed by components secreted along the oviduct. These secretion products overlay the oocytes as they transit along the different oviductal portions. In this study, we have isolated two highly glycosylated proteins of the jelly coat, which are secreted almost all the way along the oviduct. Both glycoproteins [designated as highly glycosylated protein (HGP) and low-molecular-mass highly glycosylated protein (L-HGP)] were purified to homogeneity, from the secretion of the caudal oviduct portion, by CsCl density gradient ultracentrifugation. HGP is a high-molecular-mass protein with mucin-like characteristics: high viscosity, a high content of serine and threonine, about 70% carbohydrate by weight, and a protease-resistant domain. Cleavage of disulphide bridges with reducing agents resulted in the release of a single subunit (300000 Da). L-HGP is also a disulphide-cross-linked protein with lower apparent monomeric molecular mass, in the range 100-120 kDa and containing 50% carbohydrate by weight. HGP contains galactose, fucose, N-acetylgalactosamine and sialic acid, but no mannose, suggesting the presence of O-linked oligosaccharides exclusively. The secretion ratio of HGP increases from cephalic (16% of total protein in pars preconvoluta) to caudal (40% of total protein in pars convoluta) oviductal portions. It appears to be the major structural component of the jelly coat. Our purification data suggest that HGP is non-covalently linked to the other egg jelly proteins. Polyclonal antiserum to each purified glycoprotein from secretion was raised in rabbits and used to localize both glycoproteins in the different oviductal portions, total egg jelly and the aqueous medium where oocyte strings were incubated. HGP forms a stable fibre matrix around the oocyte. L-HGP is present in the jelly coat and is released into the incubation medium.

Identification of mRNA-binding proteins during development: characterization of Bufo arenarum cellular nucleic acid binding protein.

Ultraviolet irradiation was used to covalently cross-link poly(A)+RNA and associated proteins in eggs and embryos of the toad Bufo arenarum. Four major proteins with apparent sizes of 60, 57, 45 and 30-24 kDa were identified. It was observed that the same mRNA-binding proteins were isolated from eggs to gastrula and neural stages of development. The 30 kDa polypeptide, p30, appeared as the main ultraviolet (UV) cross-linked protein in the developmental stages analyzed. By means of polyclonal antibodies, it was determined that this polypeptide has a cytoplasmic localization and it was detected in liver, eggs and embryos. The presence of p30 was also analyzed by western blot during oogenesis and development. The 30 kDa polypeptide was present in all stages analyzed but it could not be detected in stages I-II of oogenesis. At the neural stage, the relative amount of p30 began to decrease, reaching its lowest levels after stages 26-30 (tail-bud in Bufo arenarum). On the basis of purification, immunoprecipitation and western blot assays the 30 kDa protein was identified as the Bufo arenarum cellular nucleic acid binding protein.

Abordaje en el cribado de cáncer mamario en grupos poblacionales de riesgo elevado / Breast cancer screening in high risk populations

El objetivo de este trabajo es definir qué pacientes integran los grupos de riesgo alto e intermedio de padecer cáncer mamario, hacer una revisión de los estudios de cribado con resonancia magnética añadida a mamografía en pacientes de alto riesgo (y describir las características de imagen de los cánceres en este grupo), revisar los estudios con resonancia magnética realizados en subgrupos de riesgo intermedio, y realizar una actualización en recomendaciones en el cribado en pacientes con riesgo alto e intermedio (basada en las recomendaciones recientes de las principales sociedades/guías americanas y europeas)(AU)

We aim to define which patients make up the populations with high and intermediate risk of developing breast cancer, to review the studies of screening with magnetic resonance imaging in addition to mammography in high risk patients (describing the imaging characteristics of the cancers in this group), to review the studies of screening with magnetic resonance imaging in patients with intermediate risk, and to update the guidelines for screening in patients with high or intermediate risk (based on the recent recommendations of the main scientific societies/American and European guidelines)(AU)