AAV-mediated gene transfer in the perinatal period results in expression of FVII at levels that protect against fatal spontaneous hemorrhage.

We explored adeno-associated viral vector (AAV)-mediated gene transfer in the perinatal period in animal models of severe congenital factor VII (FVII) deficiency, a disease associated with early postnatal life-threatening hemorrhage. In young adult mice with plasma FVII < 1% of normal, a single tail vein administration of AAV (1 × 10(13) vector genomes [vg]/kg) resulted in expression of murine FVII at 266% ± 34% of normal for ≥ 67 days, which mediated protection against fatal hemorrhage and significantly improved survival. Codon optimization of human FVII (hFVIIcoop) improved AAV transgene expression by 37-fold compared with the wild-type hFVII cDNA. In adult macaques, a single peripheral vein injection of 2 × 10(11) vg/kg of the hFVIIcoop AAV vector resulted in therapeutic levels of hFVII expression that were equivalent in males (10.7% ± 3.1%) and females (12.3% ± 0.8%). In utero delivery of this vector in the third trimester to fetal monkeys conferred expression of hFVII at birth of 20.4% ± 3.7%, with a gradual decline to > 1% by 7 weeks. Re-administration of an alternative serotype at 12 months postnatal age increased hFVII levels to 165% ± 6.2% of normal, which remained at therapeutic levels for a further 28 weeks without toxicity. Thus, perinatal AAV-mediated gene transfer shows promise for disorders with onset of pathology early after birth.

Well-being at work and Finnish dairy farmers─from job demands and loneliness towards burnout.

Objectives: Novel information about the relationships between farmers' job demands, lack of resource, burnout, and ill health is reported based on testing the so-called "health impairment process" of the Job Demands─Resources Model (JD-R) on a representative sample of Finnish dairy farmers. The aim was to find out whether two different job demand factors; workload, societal demands and lack of resource; loneliness, were related to the indicators of ill health via burnout. Methods: The data is based on a postal survey of 400 Finnish dairy farms. Altogether 265 questionnaires were received from 188 farms and included in the analysis. The response rate was 47 per cent among sample farms. Structural equation modelling (SEM) was used to analyze the relationships between the variables. Explanatory factor analysis was used to group the job demand and lack of resource variables. Results: We identified two job demand factors, which we labelled workload and societal demands and one lacking job resource, loneliness. Our theoretical model was supported in that two of the factors, namely workload and loneliness, were related to ill health indirectly via burnout. In addition, workload was directly connected with ill health. Societal job demands were not significantly related to burnout, or to ill health. Conclusion: Our results suggest that farmers could benefit from means to reduce workload, especially the physical load. This topic needs further research as the restructuring process has increased farm enterprise sizes. There is a need to develop tools and projects to alleviate loneliness among farmers. Lack of social support, high workload, ill health, and burnout among farmers may have serious direct and indirect negative consequences for the sustainability of farming.

Evidence for a role of the NOS1AP (CAPON) gene in schizophrenia and its clinical dimensions: an association study in a South American population isolate.

BACKGROUND/AIMS: Recent studies have implicated a region on chromosome 1q21-23, including the NOS1AP gene, in susceptibility to schizophrenia. However, replication studies have been inconsistent, a fact that could partly relate to the marked psychopathological heterogeneity of schizophrenia. The aim of this study is to evaluate association of polymorphisms in the NOS1AP gene region to schizophrenia, in patients from a South American population isolate, and to assess if these variants are associated with specific clinical dimensions of the disorder. METHODS: We genotyped 24 densely spaced SNPs in the NOS1AP gene region in a schizophrenia trio sample. The transmission disequilibrium test (TDT) was applied to single marker and haplotype data. Association to clinical dimensions (identified by factor analysis) was evaluated using a quantitative transmission disequilibrium test (QTDT). RESULTS: We found significant association between eight SNPs in the NOS1AP gene region to schizophrenia (minimum p value = 0.004). The QTDT analysis of clinical dimensions revealed an association to a dimension consisting mainly of negative symptoms (minimum p value 0.001). CONCLUSIONS: Our findings are consistent with a role for NOS1AP in susceptibility to schizophrenia, especially for the 'negative syndrome' of the disorder.

Stress among Finnish farm entrepreneurs.

The aims were to examine the prevalence of stress among Finnish full-time farm entrepreneurs in 2004 (n = 1,182) and to compare the results with those for the general working population in 2003. The second aim was to analyze which factors were associated with the prevalence of stress. A stratified random sample of farm entrepreneurs gathered from the farm register was surveyed using computer-assisted telephone interviews. A binary logistic regression model was used to analyze the association with background factors. One third (34 %) of the examined farmers had experienced stress. This amount was lower than among the general working population (44 %). The most common factors associated with farmers' stress were problems in social family relationships and mental support. Physical factors such as the strenuousness of agricultural work, illness and a low estimation of their own working ability, were also related to stress. Increased stress was also associated with economic problems. Health and extension services should pay special attention to encouraging farm entrepreneurs to maintain their social relationships. The relatively low level of stress observed may indicate that those who have continued within the agricultural sector have the psychological capacity to deal with stressful situations.

Job Resources and Work Engagement among Finnish Dairy Farmers.

OBJECTIVES: The aims of this study were to examine job resources, work engagement and Finnish dairy farmers' preferences concerning methods to enhance overall well-being while working on farms. METHODS: A postal survey yielded 265 completed questionnaires from 188 dairy farms. The sample was assessed as representative of Finnish dairy farmers. Exploratory factor analysis and a linear mixed model were utilized during the data analyzing process. RESULTS: The variables lowering work engagement were stressors related to the workload and problems with health. Elevated work engagement was associated with the factors work with farm animals and family. The most important resource variables were "child or children," "own family," and "animal health." Female dairy farmers considered resource variables related to the family, love, and work with cattle as significantly more important than male dairy farmers. Male dairy farmers experienced higher work engagement and, concerning the dimensions, especially higher dedication and absorption than male respondents in a reference sample of workers in difference occupations. A sustainable farm economy and the possibility to have a holiday period were the most important methods to improve overall well-being on dairy farms. CONCLUSION: The results indicate that the family, working with cattle, healthy farm animals, a reasonable workload, and a sustainable farm economy have the capacity to create positive impacts on well-being among dairy farmers. Well-being on farms is a part of sustainable food production.

Stress and Burnout Among Finnish Dairy Farmers.

Psychosocial risks among farmers have increasingly been examined because of the ongoing changes in agriculture, such as restructuring of the industry, transition from family farming towards entrepreneurship, and climate change. The aims of the study were to determine the stressors, prevalence of stress and burnout, and variables associated with these symptoms among Finnish dairy farmers. In total 265 respondents completed a postal survey; their average age was 48 years, 44% were females and 56% males. The farms of the survey sample were larger (54 field hectares, 29 cows) than an average farm in Finland (37 hectares, 24 cows) in 2010. The most common stressors were external, such as "agricultural policy of the EU" (European Union) and "the treatment of farmers in society and the media." In addition, common stressors were related to farm and work, e.g., "amount of work," unpredictability, and "animal diseases." The prevalence of stress (42%) was found to have increased compared with earlier studies and was greater than among the general working population. All respondents as a group were classified as having slight symptoms of burnout, and one tenth (9%) of dairy farmers had experienced severe burnout. Stressors related to the workload and health were associated with stress and burnout symptoms. Also, a poor economic situation and loneliness were related to stress. Burnout correlated with a tie stall barn type and with a farm not being involved in the milk production record system. Factors protecting against burnout included positive features of the work and living environment. The study revealed changes during the past decade and new features of the well-being at work on dairy farms in Finland.

Adeno-Associated Virus (AAV) Mediated Dystrophin Gene Transfer Studies and Exon Skipping Strategies for Duchenne Muscular Dystrophy (DMD).

Duchenne muscular dystrophy (DMD), an X-linked inherited musclewasting disease primarily affecting young boys with prevalence of between1:3,500- 1:5,000, is a rare genetic disease caused by defects in the gene for dystrophin. Dystrophin protein is critical to the stability of myofibers in skeletal and cardiac muscle. There is currently no cure available to ameliorate DMD and/or its patho-physiology. A number of therapeutic strategies including molecular-based therapeutics that replace or correct the missing or nonfunctional dystrophin protein have been devised to correct the patho-physiological consequences induced by dystrophin absence. We will review the current in vivo experimentation status (including preclinical models and clinical trials) for two of these approaches, namely: 1) Adeno-associated virus (AAV) mediated (micro) dystrophin gene augmentation/ supplementation and 2) Antisense oligonucleotide (AON)-mediated exon skipping strategies.

New developments in the use of gene therapy to treat Duchenne muscular dystrophy.

INTRODUCTION: Duchenne muscular dystrophy (DMD) is a lethal X-linked inherited disorder characterised by progressive muscle weakness, wasting and degeneration. Although the gene affected in DMD was identified over 25 years ago, there is still no effective treatment. AREAS COVERED: Here we review some of the genetic-based strategies aimed at amelioration of the DMD phenotype. A number of Phase II/III clinical trials of antisense oligonucleotide-induced exon skipping for restoration of the open reading frame (ORF) of the DMD gene have recently been completed. The potential strategies for overcoming the hurdles that appear to prevent exon skipping becoming an effective treatment for DMD currently are discussed. EXPERT OPINION: The applicability of exon skipping as a therapy to DMD is restricted and the development of alternative strategies that are more encompassing is needed. The rapid pre-clinical advances that are being made in the field of adeno-associated virus (AAV)-based delivery of micro-dystrophin would address this. The obstacles to be faced with gene replacement strategies would include the need for high viral titres, efficient muscle targeting and avoidance of immune response to vector and transgene. The new emerging field of gene editing could potentially provide permanent correction of the DMD gene and the feasibility of such an approach to DMD is discussed.

Long-term episomal transgene expression from mitotically stable integration-deficient lentiviral vectors.

Nonintegrating gene delivery vectors have an improved safety profile compared with integrating vectors, but transgene retention is problematic as nonreplicating episomes are progressively and rapidly diluted out through cell division. We have developed an integration-deficient lentiviral vector (IDLV) system generating mitotically stable episomes capable of long-term transgene expression. We found that a transient cell cycle arrest at the time of transduction with IDLVs resulted in 13-45% of Chinese hamster ovary (CHO) cells expressing the transgene for over 100 cell generations in the absence of selection. The use of a scaffold/matrix attachment region did not result in improved episomal retention in this system, and episomes did not form after transduction with adeno-associated viral or minicircle vectors under the same conditions. Investigations into the episomal status of the vector genome using (1) linear amplification-mediated polymerase chain reaction followed by deep sequencing of vector-genome junctions, (2) Southern blotting, and (3) fluorescent in situ hybridization strongly suggest that the vector is not integrated in the vast majority of cells. In conclusion, we have developed an IDLV procedure generating mitotically stable episomes capable of long-term transgene expression. The application of this approach to stem cell populations could significantly improve the safety profile of a range of stem and progenitor cell gene therapies.

International perspectives on psychosocial working conditions, mental health, and stress of dairy farm operators.

Dairy farm operators-farmers, workers, and family members-are faced with many demands and stressors in their daily work and these appear to be shared across countries and cultures. Dairy operators experience high psychosocial demands with respect to a hard work and production ethos, economic influences, and social and environmental responsibility. Furthermore, both traditional and industrial farms are highly dependent on external conditions, such as weather, fluctuating markets, and regulations from government authorities. Possible external stressors include disease outbreaks, taxes related to dairy production, and recent negative societal attitudes to farming in general. Dairy farm operators may have very few or no opportunities to influence and control these external conditions, demands, and expectations. High work demands and expectations coupled with low control and lack of social support can lead to a poor psychosocial work environment, with increased stress levels, ill mental health, depression, and, in the worst cases, suicide. Internationally, farmers with ill mental health have different health service options depending on their location. Regardless of location, it is initially the responsibility of the individual farmer and farm family to handle mental health and stress, which can be of short- or long-term duration. This paper reviews the literature on the topics of psychosocial working conditions, mental health, stress, depression, and suicide among dairy farm operators, farm workers, and farm family members in an international perspective.

Microbiological quality of fresh-cut carrots and process waters.

Fresh vegetables may be contaminated by pathogens in different ways after harvest. Pathogenic microorganisms associated with fresh vegetables can cause severe outbreaks of foodborne disease. We discuss here the results of microbiological analysis of carrot samples, as well as of washing, processing, and wastewater samples. Washed, unpeeled carrots generally contained the highest aerobic plate counts (mean, 5.5 log CFU/g). Escherichia coli was not detected in any carrot or water sample examined. The amounts of coliform bacteria and enterobacteria were higher in carrot samples taken from the first steps in the processing line than in samples taken in later phases of the process. Yersinia pseudotuberculosis was not detected in any of the samples by the cultivation method; however, nonpathogenic Yersinia enterocolitica was detected from most carrot samples and almost all washing water and wastewater samples but only from 2 of 10 process water samples. Using a more-sensitive real-time PCR method, pathogenic Y. enterocolitica was found from several carrot samples, and when these positive samples were cultivated, no pathogenic Y. enterocolitica strains were detected.

Application of radiochemical determination methods in cleanability research of building materials.

During recent years increasing effort has been made to modify surface properties with easy-to-clean or self-cleaning characteristics, and concomitantly there is a need to be able to quantify cleanability. Methodology is a complex issue, including aspects of selection and characterization of the surface materials, the soiling materials (contaminants), soiling and cleaning methods, and the detection methods. Different biological, chemical, physical and visual methods have been included in studies of surface cleanability. One challenge has been to obtain quantitative information about soiling. The radiochemical methods, gamma spectrometry (NaI(Tl)-crystal) and liquid scintillation counting, have been shown to be suitable for evaluating cleanability of different surface materials and different soiling material types, providing quantitative information about the amount of soiling material both on and beneath the surface. Due to the different labelled soiling components, the interaction of the surface with different soiling material types can be evaluated. Radiochemical methods have unique benefits particularly for examining porous materials and surfaces. However, they are suitable only for highly controlled studies because of the hazards. Different features and details of radiochemical methods are discussed with the view to aid planning of future cleanability studies.

Mental symptoms among Finnish farm entrepreneurs.

The prevalence of mental symptoms among Finnish farm entrepreneurs in 2004 and 1994 was examined in two cross-sectional studies. The aim was also to examine associations between symptoms and background factors. Two random samples for computer-assisted telephone interviews comprised 1,182 full-time farmers in 2004 and 928 farmers in 1992. A binary logistic regression model was used to analyze the associations in the 2004 sample. Symptoms of weakness or fatigue and insomnia or difficulties in falling asleep were the most common symptoms in the 2004 survey, and both of these symptoms increased statistically significantly when comparing with the 1992 and 2004 surveys. Strenuousness of life and agricultural work had an association with mental symptoms. Forestry as a production sector and over two weeks of pesticide usage during the previous growing season also elevated the risk of mental symptoms. Moreover, illness or injury as well a lack of support from social relationships was associated with mental symptoms. One in four farm entrepreneurs had symptoms of weakness or fatigue in 2004. This result and the possible association between mental symptoms and pesticide usage needs special attention and further actions by health care services and other agricultural networks.