Expression analysis of candidate genes for chronic subclinical mastitis in Norwegian Red cattle.

Chronic subclinical mastitis (SCM) is characterized by a long-term inflammation in the udder with high somatic cell count (SCC) in milk. Previously, several novel alternative SCM traits for Norwegian Red (NR) cattle have been defined to improve breeding strategies against chronic SCM. Quantitative trait loci and candidate genes affecting chronic SCM in NR have been identified. The aim of this study was to analyze the expression profiles of 14 selected candidate genes (RAD17, ACOT2, ACOT4, FOS, CXCL1, CXCL8, CCNB1, CDK7, TGFB3, SEL1L, STAT4, C6, GLI2, and SLC18A2). Twenty healthy NR cows with official genomic estimated breeding values (GEBV) for lactation average somatic cell scores (LSCS) were selected. Ten cows had high GEBV for LSCS (cows with low probability to have high SCC in milk during lactation) and 10 cows had low GEBV for LSCS (cows with high probability of having high SCC in milk). We isolated RNA from unstimulated peripheral blood mononuclear cells from these. Two out of the 14 analyzed genes showed significantly different results between groups. The group with high GEBV for LSCS displayed significantly higher expression of the CXCL1 gene than the low GEBV group. Grouping by lactation stage revealed significant differential expression of the FOS gene, with higher expression in early lactation (2-3 mo after calving) compared with late lactation (7-8 mo after calving). In addition, flow cytometry was performed on the peripheral blood mononuclear cells samples to analyze if number and type of isolated cells influenced the gene expression in the groups. The results in the current study provide identified genes that can be considered as possible candidate genes for chronic SCM in NR cows.

Identification of candidate genes affecting chronic subclinical mastitis in Norwegian Red cattle: combining genome-wide association study, topologically associated domains and pathway enrichment analysis.

The aim of this study was to identify genes associated with chronic subclinical mastitis (SCM) in Norwegian Red (NR) cattle. Twelve SCM traits defined based on fixed threshold for test-day somatic cell count (SCC) were, together with lactation-average somatic cell score (LSCS) used for association and pathway enrichment analyses. A GWAS was performed on 3795 genotyped NR bulls with 777K SNP data and phenotypic information from 7 300 847 test-day SCC observations from 3 543 764 cows. At 5% chromosome-wide significance level 36 unique SNP were detected to be associated with one or more of the traits. These SNPs were analysed for linked genes using genomic positions of topologically associated domains (TAD). For the SCM traits with SCC >50 000 and >100 000 cells/ml on two test-days in a row and LSCS, the same top significant genes were identified - checkpoint clamp loader component (RAD17) and cyclin B1 (CCNB1). The SCM traits with SCC >250 000, 300 000, 350 000 or 400 000 cells/ml on two test-days in a row and D400 (number of days before the first case with SCC >400 000 cells/ml) displayed similar top significant genes: acyl-CoA thioesterase 2 and 4 (ACOT2; ACOT4). For the traits SCM200_3 (SCC >200 000 cells/ml on three test-days in a row) and SCM150, SCM200 (SCC >150 000; 200 000 cells/ml on two test-days in a row) a group of chemokine (C-X-C motif) ligand genes and the Fos proto-oncogene, AP-1 transcription factor subunit (FOS) gene, were identified. Further functional studies of these identified candidate genes are necessary to clarify their actual role in development of chronic SCM in NR cattle.

Alternative subclinical mastitis traits for genetic evaluation in dairy cattle.

Chronic subclinical mastitis (SCM), characterized by changes in milk composition and high somatic cell count (SCC) in milk for a prolonged period of time, is often caused by a bacterial infection. Different levels of SCC have been suggested and used as threshold to identify subclinical infection. The aim of this study was to examine different definitions of SCM based on test-day SCC and estimate genetic parameters for these traits and their genetic correlation to milk production. Test-day SCC records from 1,209,128 Norwegian Red cows in lactation 1 to 3 were analyzed. Twelve SCM traits were defined as binary with 2 test-day SCC in a row above SCC thresholds from 50,000 to 400,000 cells/mL (SCM50, SCM100, SCM150, SCM200, SCM250, SCM300, SCM350, and SCM400), with 3 test-day SCC in a row above 200,000 and 400,000 cells/mL (SCM200_3 and SCM400_3), and the number of days before the first case with SCM50 (D50) or SCM400 (D400). The heritability and genetic correlations were estimated for SCM traits and the mean lactation-average somatic cell score (LSCS) using linear animal repeatability models. The total mean frequency of SCM ranged from 1.2% to 51.8%, for different trait definitions, high for low SCC threshold (SCM50) and low for the highest SCC threshold (SCM400_3). For the 2 traits based on number of days, the mean values were 104 (D50) and 117 (D400) days. The mean LSCS was 4.4 (equivalent to around 82,000 SCC). Heritabilities for the 12 alternative SCM traits were low and varied from 0.01 (SCM400_3) to 0.1 (SCM100), whereas for LSCS the estimated heritability was 0.3 and standard error varied from 0.001 to 0.003. Genetic correlations among the SCM traits ranged from 0.7 (D50 and SCM400) to 1 (SCM350 and SCM400), whereas between SCM traits and milk production the correlation ranged from 0.07 (LSCS) to 0.3 (D400). The standard error for genetic correlations varied from 0.001 to 0.06. The heritability was low and the genetic correlations were strong among SCM traits. Genetic correlations lower than 1 suggest that the alternative SCM traits are genetically different from LSCS, the trait currently used in genetic evaluations for Norwegian Red. Hence, the alternative traits will add information and improve breeding for better udder health.

The dynamic changes of dominant clones of Staphylococcus aureus causing bloodstream infections in the European region: results of a second structured survey.

Staphylococcus aureus is one of the most important human pathogens and meticillin-resistant S. aureus (MRSA) presents a major cause of healthcare- and community-acquired infections. This study investigated the spatial and temporal changes of S. aureus causing bacteraemia in Europe over a five-year interval and explored the possibility of integrating pathogen-based typing data with epidemiological and clinical information at a European level. Between January 2011 and July 2011, 350 laboratories serving 453 hospitals in 25 countries collected 3,753 isolates (meticillin-sensitive S. aureus (MSSA) and MRSA) from patients with S. aureus bloodstream infections. All isolates were sent to the national staphylococcal reference laboratories and characterised by quality-controlled spa typing. Data were uploaded to an interactive web-based mapping tool. A wide geographical distribution of spa types was found, with some prevalent in all European countries. MSSA was more diverse than MRSA. MRSA differed considerably between countries with major international clones expanding or receding when compared to a 2006 survey. We provide evidence that a network approach of decentralised typing and visualisation of aggregated data using an interactive mapping tool can provide important information on the dynamics of S. aureus populations such as early signalling of emerging strains, cross-border spread and importation by travel.

Level of C - reactive protein as an indicator for prognosis of premature uterine contractions.

BACKGROUND AND OBJECTIVES: high concentrations of maternal C-reactive protein have been associated with adverse pregnancy outcome, and premature uterine contraction may be predicted by elevated levels of C-reactive protein. This may ultimately be simple and cost-effective enough to introduce as a low-risk screening program. PATIENTS AND METHODS: an observational case control study was performed from May 1st, 2010 to December 1st, 2010 at Maternity Teaching Hospital-Erbil/ Kurdistan Region/ Iraq. The sample size was (200) cases. Hundred of them were presented with premature uterine contractions at 24(+0)-36(+6) weeks. The other hundred were control group at same gestational ages. The level of C-reactive protein was determined in both groups and both groups were followed till delivery. RESULTS: (93) out of (100) women with premature uterine contractions had elevated level of C-Reactive protein and 91% delivered prematurely while in the control group only (9) out of (100) women had elevated level of C-reactive protein and only 8% of them delivered preterm. Differences were statistically highly significant. CONCLUSION: C-reactive protein can be used as a biomarker in prediction of premature delivery when it is associated with premature uterine contractions. As well it can be used as a screening test to detect cases that are at risk of premature delivery.

Embarazadas con lupus eritematoso sistémico en el hospital Clínico Regional de Concepción, Chile / Pregnant women with systemic lupus erythematosus in the Regional Clinical Hospital of Concepcion, Chile

Antecedentes: El lupus eritematoso sistémico (LES) afecta principalmente a mujeres en edad fértil. El embarazo en estas pacientes puede asociarse con múltiples complicaciones. Objetivo: Caracterizar a las embarazadas con LES durante 10 años en el Hospital Clínico Regional de Concepción. Métodos: Se realizó un estudio descriptivo retrospectivo que consistió en la revisión de fichas clínicas. Se analizaron las variables: edad, años de enfermedad desde el diagnóstico, historia obstétrica, presencia de reactivaciones, anticuerpos maternos y complicaciones materno-fetales. Resultados: Durante el periodo de estudio hubo 49 embarazos en 21 pacientes con LES. El 12,2 por ciento terminó en aborto, un 2 por ciento en óbitos, y un total de 43 nacidos vivos. La edad promedio de las pacientes al momento del diagnóstico de LES fue 24,5 años. El 67 por ciento fueron diagnosticadas antes de su primer embarazo. En el total de pacientes el 85,7 por ciento presentaron ANA positivo, 57,1 por ciento antiDNA positivo, 52,4 por ciento aRo positivo y 33,3 por ciento aLa positivo. En los caso de abortos, aRo y aLa se encontraban positivos en 66,7 por ciento. Las anticardiolipinas se encontraban alteradas en 33,3 por ciento de los abortos. Durante el embarazo el 32,6 por ciento tenía LES activo y 34,7 por ciento en el postparto. El 53,5 por ciento de los recién nacidos no tuvieron complicaciones. La complicación más frecuente fue la prematuridad con 55 por ciento. La mortalidad perinatal de la serie fue de 46,5/1000 nacidos vivos (2/43). No hubo muertes maternas. Conclusión: Es importante la educación respecto al embarazo en pacientes con LES. Debemos resaltar en promover que estas pacientes planifiquen el embarazo en periodo de inactividad, y con controles frecuentes para pesquisar precozmente cualquier complicación.

Background: The systemic lupus erythematosus (SLE) affects mainly fertile age women. Pregnancy in these patients can associate with multiple complications. Aims: To characterize the pregnant women with SLE during 10 years in the Hospital Clínico Regional de Concepción, Chile. Methods: We made a retrospective descriptive study which consisted in clinical files revision. The following variables were analyzed: age, years with disease since diagnose, obstetric history, history of reactivation, maternal antibodies and mother-fetus complications. Results: During the time of study there were 49 pregnancies on 21 patients with SLE; 12.2 percent ended in abortion, 2 percent in late fetal death giving a total of 43 living newborn. The average age of these patients at the moment of diagnose of LES was 24.5 years old; 67 percent were diagnosed before their first pregnancy. From the total of patients, 85.7 percent presented positive ANA, 57.1 percent positive antiDNA, positive aRo in 52.4 percent and positive aLa in 33.3 percent. In case of abortions, aRo and aLa were positive in 66.7 percent. Anticardiolipins were altered in 33.3 percent of abortions. During pregnancy 32.6 percent had active SLE, and 34.7 percent post-partum. Among the newborn, 53.5 percent did not have any complications. The most frequent complication was prematurity with a 55 percent. The perinatal mortality was 46.5/1000 lives births (2/43). There were no maternal deaths. Conclusion: It is important to educate about pregnancy in SLE patients. We must emphasize to promote in those patients a planned pregnancy in inactive period and with frequent controls for early diagnose of any complication.

Overview of molecular typing methods for outbreak detection and epidemiological surveillance.

Typing methods for discriminating different bacterial isolates of the same species are essential epidemiological tools in infection prevention and control. Traditional typing systems based on phenotypes, such as serotype, biotype, phage-type, or antibiogram, have been used for many years. However, more recent methods that examine the relatedness of isolates at a molecular level have revolutionised our ability to differentiate among bacterial types and subtypes. Importantly, the development of molecular methods has provided new tools for enhanced surveillance and outbreak detection. This has resulted in better implementation of rational infection control programmes and efficient allocation of resources across Europe. The emergence of benchtop sequencers using next generation sequencing technology makes bacterial whole genome sequencing (WGS) feasible even in small research and clinical laboratories. WGS has already been used for the characterisation of bacterial isolates in several large outbreaks in Europe and, in the near future, is likely to replace currently used typing methodologies due to its ultimate resolution. However, WGS is still too laborious and time-consuming to obtain useful data in routine surveillance. Also, a largely unresolved question is how genome sequences must be examined for epidemiological characterisation. In the coming years, the lessons learnt from currently used molecular methods will allow us to condense the WGS data into epidemiologically useful information. On this basis, we have reviewed current and new molecular typing methods for outbreak detection and epidemiological surveillance of bacterial pathogens in clinical practice, aiming to give an overview of their specific advantages and disadvantages.

Bioinformatics in bacterial molecular epidemiology and public health: databases, tools and the next-generation sequencing revolution.

Advances in typing methodologies have been the driving force in the field of molecular epidemiology of pathogens. The development of molecular methodologies, and more recently of DNA sequencing methods to complement and improve phenotypic identification methods, was accompanied by the generation of large amounts of data and the need to develop ways of storing and analysing them. Simultaneously, advances in computing allowed the development of specialised algorithms for image analysis, data sharing and integration, and for mining the ever larger amounts of accumulated data. In this review, we will discuss how bioinformatics accompanied the changes in bacterial molecular epidemiology. We will discuss the benefits for public health of specialised online typing databases and algorithms allowing for real-time data analysis and visualisation. The impact of the new and disruptive next-generation sequencing methodologies will be evaluated, and we will look ahead into these novel challenges.

Combining quantitative trait loci and heterogeneous microarray data analyses reveals putative candidate pathways affecting mastitis in cattle.

Mastitis is a frequent disease and considerable problem for the global dairy industry. Identification of solutions leading to the development of new control strategies is therefore of high importance. In this study, we have integrated genomic data from genome-wide association mapping in cattle with transcriptomic data from microarray studies of several mastitis pathogens and host species in vitro and in vivo. To identify significant candidate pathways directly and indirectly involved in the immune response to mastitis, ingenuity pathway analysis (ipa) and database for annotation, visualization and integrated discovery bioinformatic (david) were applied. Several candidate pathways were found. Of great interest are IL-17 and IL-8 signalling pathways, responsible for the recruitment and migration of inflammatory cells into tissue during inflammation and infection. These results may emphasize further functional studies for identification of factors contributing to resistance to mastitis pathogens in cattle.

Prevalencia de dislipidemias en los pacientes con lupus eritematoso sistémico en el Hospital Clínico Regional de Concepción / Prevalence of dyslipidemia in patients with systemic lupus erythematosus in Hospital Clínico Regional of Concepción

INTRODUCCIÓN: El lupus eritematoso sistémico es considerado un factor de riesgo cardiovascular. En estos pacientes es frecuente hallar dislipidemias. La terapia corticoesteroide favorece las alteraciones lipídicas. HIPÓTESIS: Los pacientes con lupus eritematoso sistémico presentan hipercolesterolemia, hipertrigliceridemia o ambos, en mayor proporción que la población general. OBJETIVO: Determinar la prevalencia de dislipidemias en los pacientes diagnosticados de lupus eritematoso sistémico y controlados en el policlínico de reumatología del Hospital Clínico Regional de Concepción. MATERIAL Y MÉTODO: Estudio descriptivo transversal en pacientes diagnosticados de lupus eritematoso sistémico, y que se han controlado durante el 2010 en el policlínico de reumatología. Las variables estudiadas fueron: sexo, edad, hipercolesterolemia, hipertrigliceridemia, dosis/ tiempo de uso de corticoides e hidroxicloroquina. RESULTADOS: Total de pacientes estudiados (n=154), 88,9 por ciento (n=137) mujeres, edad promedio45,6 + 5 años, 40,9 por ciento (n=43) presenta hipertrigliceridemia; 55,2 por ciento (n=58) presenta hipercolesterolemia y 65,9 por ciento (n=100) presenta ambas alteraciones. Pacientes con evolución menor de 10 años: 50 por ciento (n=29) presenta hipercolesterolemia, 34,5 por ciento (n=20) presentaba hipertrigliceridemia; con 11-20 años de evolución: 56,25 por ciento (n=18) presenta hipercolesterolemia.; 43,75 por ciento (n=14) presenta hipertrigliceridemia Con evolución mayor a 20 años: 73,3 por ciento (n=11) presenta hipercolesterolemia; 60 por ciento (n=9) presentaba hipertrigliceridemia Usuarios de corticoides 89 por ciento (n=138) y de hidroxicloroquina 92,4 por ciento (n=142). DISCUSIÓN: La principal alteración es hipercolesterolemia. Pacientes con más años de evolución presentan mayor número de dislipidemias. La mayoría son usuarios de corticoides. Concluimos que la prevalencia de...

INTRODUCTION: Systemic lupus erythematosus is considered a cardiovascular risk factor. Dyslipidemia is commonly found in these patients. Corticosteroid therapy promotes lipid abnormalities. HYPOTHESES: Systemic lupus erythematosus patients with hypercholesterolemia, hypertriglyceridemia, or both, in greater proportion than the general population. OBJECTIVE: To determine the prevalence of dyslipidemia in patients diagnosed with systemic lupus erythematosus and monitored at the clinic of rheumatology, Hospital Clínico Regional Concepción. MATERIAL AND METHOD: A retrospective cross sectional in patients diagnosed with systemic lupus erythematosus and have controlled the last year in the Hospital Clínico Regional Concepción. The variables studied were: sex, age, hypercholesterolemia, hypertriglyceridemia, dose/duration of use of corticosteroids and hydroxichloroquine. RESULTS: Of all patients studied (n=154), 88.9 percent (n=137) women, mean age 45.6 +/- 5 years, 40.9 percent (n=43) presents hypertriglyceridemia, 55.2 percent (n=58) presented hypercholesterolemia presented and 65.9 percent (n=100) presented both alterations. Patients with minor changes 10 years: 50 percent (n=29) presents hypercholesterolemia; 34.5 percent (n=20) had hypertriglyceridemia. With11-20 years of evolution: 56.25 percent (n=18) presents hypercholesterolemia;45.75 percent (n=14) has hypertriglyceridemia. With over 20years of evolution: 73.3 percent (n=11) has hypercholesterolemia; 60 percent(n=9) showed hypertriglyceridemia. Steroids users 89 percent (n=138)and 92.4 percent (n=142) using hixychloroquine. DISCUSSION: The main alteration is hypercholesterolemia. Patients with more years of evolution have a greater number of dyslipidemia than others patients. Most are steroid users. We conclude that the prevalence of dyslipidemia is more common in systemic lupus erythematosus patients than the general population...

Extremely low genetic variability and highly structured local populations of Arabidopsis thaliana at higher latitudes.

The genetic diversity and population structure of Arabidopsis thaliana populations from Norway were studied and compared to a worldwide sample of A. thaliana to investigate the demographic history and elucidate possible colonization routes of populations at the northernmost species limit. We genotyped 282 individuals from 31 local populations using 149 single nucleotide polymorphism markers. A high level of population subdivision (F(ST) = 0.85 ± 0.007) was found indicating that A. thaliana is highly structured at the regional level. Significant relationships between genetic and geographical distances were found, suggesting an isolation by distance mode of evolution. Genetic diversity was much lower, and the level of linkage disequilibrium was higher in populations from the north (65-68°N) compared to populations from the south (59-62°N); this is consistent with a northward expansion pattern. A neighbour-joining tree showed that populations from northern Norway form a separate cluster, while the remaining populations are distributed over a few minor clusters. Minimal gene flow seems to have occurred between populations in different regions, especially between the geographically distant northern and southern populations. Our data suggest that northern populations represent a homogenous group that may have been established from a few founders during northward expansions, while populations in the central part of Norway constitute an admixed group established by founders of different origins, most probably as a result of human-mediated gene flow. Moreover, Norwegian populations appeared to be homogenous and isolated compared to a worldwide sample of A. thaliana, but they are still grouped with Swedish populations, which may indicate common colonization histories.

Two allelic forms of the aureolysin gene (aur) within Staphylococcus aureus.

Proteinases of Staphylococcus aureus are emerging as potential virulence factors which may be involved in the pathogenecity of staphylococcal diseases. We describe here the structure of the gene encoding the metalloproteinase referred to as aureolysin. This gene occurs in two allelic forms and is strongly conserved among S. aureus strains, implying the possibility that the proteinase may have important housekeeping functions.

Seedling establishment in an epiphytic orchid: an experimental study of seed limitation.

Will increased fruit and seed production in a severely pollination-limited orchid stimulate population growth? We tested whether safe sites for germination and seedling establishment are limiting for the twig epiphyte, Tolumnia variegata, by manipulating fruit set and monitoring subsequent seedling establishment for two seasons (1991-1992, 1992-1993). In the Cambalache Forest Reserve of Puerto Rico, we established 36 plots along a transect. Each plot consisted of nine trees. A center tree was designated as the site for attaching Tolumnia and manipulating fruit set. The other eight potential host trees were 1-3 and 3-5 m from the center tree in each of the cardinal directions. A 1-m length of stem 1 m from the ground was monitored for recruits on each of the nine trees of 24 fruit-enhanced plots and 12 controls (23 and 13, respectively for the 1992-1993 season). Fruit enhancement plots were divided among two treatments: one-fruit and five-fruit additions for the 1st year and one to five and more than five fruits for the 2nd year. Availability of suitable host species was not limiting. T. variegata showed little specificity for host tree species, good host trees and shrubs were common, and there was no evidence that the orchid had a preference for small branches, despite possessing the entire suite of characteristics thought to respresent "obligate" twig epiphytes. Fruit enhancement increased seed rain and seedling establishment consistently in only the high-fruit treatment plots. Most recruitment occurred near fruiting plants. Over the 2-year period, mortality was 18% for adults and 85.5% for the 1991-1992 cohort of recruits. Net recruitment was positive for both the treatment (average = 1.74) and control plots (average = 0.67). Seedling establishment at our study site was not microsite-limited. If selection for increased pollinator attraction occurs, then an increase in seed output should result in population growth.