RICERCA E VALIDAZIONE DI SNP IN GENI CANDIDATI PER LA QUALIT\uc0 DELLA CARNE E APPLICAZIONE DELL'ANALISI GENOMICA ALLA SPECIE SUINA

An important aim of pig selection in Italy is to obtain animals having a high aptitude for the PDO dry-cured ham production, such as Parma or S. Daniele ham. Over the past years, advances in the porcine genetic map have led to valuable gene and trait information being discovered. Since that time, sequences for the pig genome have been generated from various tissues, the sequencing of candidate genes, and more recently large scale genomic sequencing projects. These efforts are also being directed to SNPs identification for future large scale association studies. In the next years, the efficiency and accuracy of the traditional pig selection schemes could be improved by the implementation of molecular data into breeding programs. In this work, seven swine candidate genes for meat quality were investigated in order to identify informative SNPs. Molecular analyses were performed on twenty-two animals representing the extreme tails of the Gaussian distribution for three selected phenotypes (muscle compactness, fat thickness and the principal component 1) of 231 Large White x Landrace individuals. Among the nine identified SNPs, only two SNPs in the CRADD gene, two SNPs in the PTPRD gene and one SNP in the PIK3R2 gene showed a MAF (Minor Allele Frequency) more than 5% in the animals tested and therefore were considered for the subsequent association analysis. Association analysis between these five SNPs and the three phenotypes considered in this study was carried out using the GML procedure. The SNP CRADD g343 [A/G] showed a good association with the compactness of the muscles (P = 0,0498), the SNP PTPRD g30194 [G/T] showed a good association with the compactness of the muscles (P = 0,0195) and fat thickness dorsal (P = 0,0265), the SNP PIK3R2 g.3008 [C/T] showed a very good association with the compactness of muscle mass (P = 00,0014) and thickness of backfat (P = 0,0087). Therefore, the PIK3R2SNP g.3008 [C / T] was genotyped on all 231 animals of the population. The analysis showed a significant effect of this SNP on the following variables: marbling (P < 0.0001), fat cover (P < 0.05), fat thickness (P < 0.05), Prin1 (P < 0.05), Prin3 (P < 0.01); Prin4 (P < 0.01). In particular, the CC genotype was positively associated with marbling and fat cover. Moreover, the SNP in PIK3R2 gene was tested on 600 samples of three different Italian breeds (Large White, Duroc, Landrace) obtained from the National Association of Pig Breeders of Italy. Within each breed, the 100 individuals with the highest and the 100 individuals with the lowest values for EBVs (Estimated Breeding Values) for average daily weight gain were analyzed, resulting this SNP polymorphic in each breed. The association analyses between this SNP and these extreme EBVs showed a good association with backfat thikness, average daily gain, feed conversion rate and thigh weight in Landrace individuals. The two SNPs identified in CRADD gene and the two SNPs identified in the PTPRD gene were tested in another group of 560 Italian Large White animals with extreme EBVs for fat thickness. Three of the four SNPs resulted polymorphic also in this population. Then, association analysis between these three SNPs and EBVs for fat thickness were performed, showing the association of the SNP g29962[A/G ] of PTPRD gene with thigh weight. Another aim of this PhD thesis is the genetic characterization of a swine genetic type, the \u201cNero di Garlasco\u201d, expressing both ancient and recent biodiversity. Although in the last years admirable efforts have been made to recover pig biodiversity, extremely endangered, today only few Italian local breeds can withstand the competition with commercial foreign breeds (i.e. the Large White, Landrace and Duroc) and with the commercial crosses today more and more widespread in the market. To date, at the national Herd Book only few breeds are registered as the Cinta senese, Mora Romagnola, Nero Siciliano, Casertana, Apulo-Calabrese and the Sarda. A very limited number with respect to the tens of breeds and lines cited in the textbooks of agronomy of the second postwar period. The majority of the extinct pig breeds were adapted to free range breeding, and were characterized by dark coat color (to defend themselves form the sun), slow growth and extensive fat deposits. In recent years, an ancient genetic type barely still existing (Razza di Garlasco) is reconstructing in the province of Pavia (Lomellina). In this case, pig phenotypes consisted of dark coat color and, surprisingly, high growth rates, similar to the commercial breeds. These two characteristics made these animals well adapted for the non-industrial production, where animals can be bred in free range, because the dark color protects them from the sun. To achieve this goal, a genetic characterization of the model population should be conducted. The tight bottleneck, through which this small population has passed, allows the fixation of genetic markers that enable precise traceability of the fresh and transformed products. This can give an added value to the breeding of theses animals that responds to the request for security and sustainability of the animal production system. Thanks to innovative technologies, a population of 96 animals, belonging to the Nero di Garlasco breed, was analyzed using the PorcineSNP60 BeadChip, in order to screen about 60,000 SNPs. The obtained data allowed a first description of the genetic structure of this population, but further studies are required to characterize this swine genetic type

Risk of Guillain-Barré syndrome after 2010–2011 influenza vaccination

Influenza vaccination has been implicated in Guillain Barré Syndrome (GBS) although the evidence for this link is controversial. A case–control study was conducted between October 2010 and May 2011 in seven Italian Regions to explore the relation between influenza vaccination and GBS. The study included 176 GBS incident cases aged ≥18 years from 86 neurological centers. Controls were selected among patients admitted for acute conditions to the Emergency Department of the same hospital as cases. Each control was matched to a case by sex, age, Region and admission date. Two different analyses were conducted: a matched case–control analysis and a self-controlled case series analysis (SCCS). Case–control analysis included 140 cases matched to 308 controls. The adjusted matched odds ratio (OR) for GBS occurrence within 6 weeks after influenza vaccination was 3.8 (95 % CI: 1.3, 10.5). A much stronger association with gastrointestinal infections (OR = 23.8; 95 % CI 7.3, 77.6) and influenza-like illness or upper respiratory tract infections (OR = 11.5; 95 % CI 5.6, 23.5) was highlighted. The SCCS analysis included all 176 GBS cases. Influenza vaccination was associated with GBS, with a relative risk of 2.1 (95 % CI 1.1, 3.9). According to these results the attributable risk in adults ranges from two to five GBS cases per 1,000,000 vaccinations

Genomic characteristics of Staphylococcus aureus strains associated with high within-herd prevalence of intramammary infections in dairy cows

Staphylococcus aureus is one of the most important causes of mastitis in dairy cattle. Based on previous research, Staph. aureus genotypes with different pathogenic and contagious properties can cause intramammary infection (IMI) and coexist in the same herd. Our study aimed to compare Staph. aureus strains from herds that differed in IMI prevalence using different molecular approaches such as ribosomal spacer (RS)-PCR, multilocus sequence typing (MLST), spa typing, ribotyping, pulsed-field gel electrophoresis (PFGE), and multiplex PCR. For this purpose, 31 dairy herds with Staph. aureus IMI were selected, and 16 of these were chosen for a comparison study: the 8 high-prevalence (HP) herds had Staph. aureus IMI prevalence >28% and the 8 low-prevalence (LP) herds had an IMI prevalence <4%. A total of 650 isolates of Staph. aureus from mammary quarters of all positive cows were genotyped with RS-PCR, a technique based on amplification of a portion of the intergenic spacer 16S-23S rRNA, and a subset of 54 strains was also analyzed by multiplex PCR, ribotyping, PFGE, MLST, and spa typing. The RS-PCR analysis revealed 12 different profiles. Staphylococcus aureus strains isolated from 5 out of 8 HP herds showed a profile identical to the genotype B (GTB), described in previous studies as being strongly associated with high within-herd prevalence of Staph. aureus mastitis and the presence of the genes coding for enterotoxins sea, sed, and sej, a long x-region of spa gene, and 3 lukE fragments. Moreover, all strains isolated in the HP herds possessed genes coding for staphylococcal enterotoxins. In LP herds, a limited number of strains of 6 genotypes, different from those isolated in HP herds, were identified and GTB was not found. Within these genotypes, 4 strains were positive for the mecA gene. Preliminary results and comparison with other genotyping methods confirmed that genotyping by RS-PCR is an accurate, rapid, and inexpensive tool for future field studies on Staph. aureus mastitis strains and generates clinically relevant results

Ultrasound-assessed lung aeration correlates with respiratory system compliance in adults and neonates with acute hypoxemic restrictive respiratory failure: an observational prospective study

Background: Lung ultrasound allows lung aeration to be assessed through dedicated lung ultrasound scores (LUS). Despite LUS have been validated using several techniques, scanty data exist about the relationships between LUS and compliance of the respiratory system (Crs) in restrictive respiratory failure. Aim of this study was to investigate the relationship between LUS and Crs in neonates and adults affected by acute hypoxemic restrictive respiratory failure, as well as the effect of patients' age on this relationship. Methods: Observational, cross-sectional, international, patho-physiology, bi-center study recruiting invasively ventilated, adults and neonates with acute respiratory distress syndrome (ARDS), neonatal ARDS (NARDS) or respiratory distress syndrome (RDS) due to primary surfactant deficiency. Subjects without lung disease (NLD) and ventilated for extra-pulmonary conditions were recruited as controls. LUS, Crs and resistances (Rrs) of the respiratory system were measured within 1 h from each other. Results: Forty adults and fifty-six neonates were recruited. LUS was higher in ARDS, NARDS and RDS and lower in control subjects (overall p &lt; 0.001), while Crs was lower in ARDS, NARDS and RDS and higher in control subjects (overall p &lt; 0.001), without differences between adults and neonates. LUS and Crs were correlated in adults [r = - 0.86 (95% CI - 0.93; - 0.76), p &lt; 0.001] and neonates [r = - 0.76 (95% CI - 0.85; - 0.62), p &lt; 0.001]. Correlations remained significant among subgroups with different causes of respiratory failure; LUS and Rrs were not correlated. Multivariate analyses confirmed the association between LUS and Crs both in adults [B = - 2.8 (95% CI - 4.9; - 0.6), p = 0.012] and neonates [B = - 0.045 (95% CI - 0.07; - 0.02), p = 0.001]. Conclusions: Lung aeration and compliance of the respiratory system are significantly and inversely correlated irrespective of patients' age. A restrictive respiratory failure has the same ultrasound appearance and mechanical characteristics in adults and neonates

Effects of n-3 fatty acids, EPA v. DHA, on depressive symptoms, quality of life, memory and executive function in older adults with mild cognitive impairment: a 6-month randomised controlled trial

First published online 20 September 2011Depressive symptoms may increase the risk of progressing from mild cognitive impairment (MCI) to dementia. Consumption of n-3 PUFA may alleviate both cognitive decline and depression. The aim of the present study was to investigate the benefits of supplementing a diet with n-3 PUFA, DHA and EPA, for depressive symptoms, quality of life (QOL) and cognition in elderly people with MCI. We conducted a 6-month double-blind, randomised controlled trial. A total of fifty people aged >65 years with MCI were allocated to receive a supplement rich in EPA (1·67 g EPA + 0·16 g DHA/d; n 17), DHA (1·55 g DHA + 0·40 g EPA/d; n 18) or the n-6 PUFA linoleic acid (LA; 2·2 g/d; n 15). Treatment allocation was by minimisation based on age, sex and depressive symptoms (Geriatric Depression Scale, GDS). Physiological and cognitive assessments, questionnaires and fatty acid composition of erythrocytes were obtained at baseline and 6 months (completers: n 40; EPA n 13, DHA n 16, LA n 11). Compared with the LA group, GDS scores improved in the EPA (P=0·04) and DHA (P=0·01) groups and verbal fluency (Initial Letter Fluency) in the DHA group (P=0·04). Improved GDS scores were correlated with increased DHA plus EPA (r 0·39, P=0·02). Improved self-reported physical health was associated with increased DHA. There were no treatment effects on other cognitive or QOL parameters. Increased intakes of DHA and EPA benefited mental health in older people with MCI. Increasing n-3 PUFA intakes may reduce depressive symptoms and the risk of progressing to dementia. This needs to be investigated in larger, depressed samples with MCI.Natalie Sinn, Catherine M. Milte, Steven J. Street, Jonathan D. Buckley, Alison M. Coates, John Petkov, and Peter R. C. How

Recurrence of Cardiovascular Events in Patients With Type 2 Diabetes : Epidemiology and risk factors

OBJECTIVE—The purpose of this study was to assess incidence of and risk factors for recurrent cardiovascular disease (CVD) in type 2 diabetes

Pharmacotherapy negative outcomes resulting in Primary Care Emergency visits

Objetivo Analizar la prevalencia de los resultados negativos asociados a la medicación (RNM) que son causa de consulta en un servicio de urgencias de atención primaria (SUAP) en un entorno rural. Determinar la evitabilidad y la gravedad de los mismos. Diseño Estudio observacional descriptivo transversal. Emplazamiento SUAP de Mula. Murcia. Participantes Un total de 330 pacientes, en un periodo de 33 semanas. Mediciones principales Número y tipo de RNM: el farmacéutico, a través de los datos obtenidos de un cuestionario validado y la historia clínica, evaluó si existía relación entre los medicamentos que toma el paciente y el motivo de acudir a urgencias. En caso de sospecha de RNM se reevaluaba con el médico y se confirmaban o no los RNM identificados. Resultados De los 330 pacientes fueron evaluables 317. La media de edad de los pacientes era de 39,63 años y el 51,42% eran mujeres. La media de medicamentos que utilizaban fue de 1,38. Se detectaron un 26,50% (IC 95% 21,94-31,62) de pacientes con RNM como causa de visita a urgencias. El 53,57% de los RNM detectados fue de la categoría de efectividad y el 40,48% de necesidad. El 77,41% (IC-95% 67,35-85,01) de las visitas causadas por RNM fueron evitables. En cuanto a la gravedad, el 92,986% de los RNM eran leves. Conclusiones Una de cada 4 visitas al SUAP de Mula está causada por un RNM y, de ellas, el 77,41% son evitables.Objective Our aim was to estimate the prevalence of Pharmacotherapy negative outcomes in Primary Care Emergency visits in a rural environment, and to determine their preventability and severity. Design Descriptive study with an analytical component. Site Primary Care Emergency Service (SUAP), Mula. Murcia. Patients The study consisted of 330 patients over a 33 week period. Method Number and type of Pharmacotherapy negative outcomes: Pharmacist through the data, a validated questionnaire and medical history, assessing whether there was a relationship between the medications and the patient, and the reason for going to the Primary Care Emergency. In case of suspicion of Pharmacotherapy negative outcomes the patient is reassessed by the doctor, and the Pharmacotherapy negative outcomes confirmed or not identified. Results Of the 330 patients, 317 were evaluable. The mean age of patients was 39.63 years and 51.42% were women. The mean number of drugs used was 1.38, and 26.50% (95% CI, 21.94% -31.62%) patients were detected with Pharmacotherapy negative outcomes as a cause of visiting the Primary Care Emergency. 53.57% of the detected Pharmacotherapy negative outcomes detected as regards efficacy was 53.75%, 40.48% as regards need. More than three-quarters (77.41%; 95% CI, 67.35% -85.01%) of emergency visits caused by Pharmacotherapy negative outcomes were avoidable. In terms of severity, 92.86% of the Pharmacotherapy negative outcomes were mild. Conclusions One in four Mula SUAP visits are due to a Pharmacotherapy negative outcomes, and 77.41% of them are preventable

Rat models of acute inflammation: a randomized controlled study on the effects of homeopathic remedies

BACKGROUND: One of the cardinal principles of homeopathy is the "law of similarities", according to which patients can be treated by administering substances which, when tested in healthy subjects, cause symptoms that are similar to those presented by the patients themselves. Over the last few years, there has been an increase in the number of pre-clinical (in vitro and animal) studies aimed at evaluating the pharmacological activity or efficacy of some homeopathic remedies under potentially reproducible conditions. However, in addition to some contradictory results, these studies have also highlighted a series of methodological difficulties. The present study was designed to explore the possibility to test in a controlled way the effects of homeopathic remedies on two known experimental models of acute inflammation in the rat. To this aim, the study considered six different remedies indicated by homeopathic practice for this type of symptom in two experimental edema models (carrageenan- and autologous blood-induced edema), using two treatment administration routes (sub-plantar injection and oral administration). METHODS: In a first phase, the different remedies were tested in the four experimental conditions, following a single-blind (measurement) procedure. In a second phase, some of the remedies (in the same and in different dilutions) were tested by oral administration in the carrageenan-induced edema, under double-blind (treatment administration and measurement) and fully randomized conditions. Seven-hundred-twenty male Sprague Dawley rats weighing 170–180 g were used. Six homeopathic remedies (Arnica montana D4, Apis mellifica D4, D30, Atropa belladonna D4, Hamamelis virginiana D4, Lachesis D6, D30, Phosphorus D6, D30), saline and indomethacin were tested. Edema was measured using a water-based plethysmometer, before and at different times after edema induction. Data were analyzed by ANOVA and Student t test. RESULTS: In the first phase of experiments, some statistically significant effects of homeopathic remedies (Apis, Lachesis and Phosporus) were observed (the reduction in paw volume increase ranging from 10% to 28% at different times since edema induction). In the second phase of experiments, the effects of homeopathic remedies were not confirmed. On the contrary, the unblinded standard allopathic drug indomethacin exhibited its anti-inflammatory effect in both experimental phases (the reduction in paw volume increase ranging from 14% to 40% in the first phase, and from 18% to 38% in the second phase of experiments). CONCLUSION: The discrepancies between single-blind and double-blind methods in animal pharmacological research are noteworthy and should be better investigated, also in non-homeopathic research