Safe and cost-effective protocol for shipment of samples from Foot-and-Mouth Disease suspected cases for laboratory diagnostic.

An essential step towards the global control and eradication of foot-and-mouth disease (FMD) is the identification of circulating virus strains in endemic regions to implement adequate outbreak control measures. However, due to the high biological risk and the requirement for biological samples to be shipped frozen, the cost of shipping samples becomes one of major obstacles hindering submission of suspected samples to reference laboratories for virus identification. In this study, we report the development of a cost-effective and safe method for shipment of FMD samples. The protocol is based on the inactivation of FMD virus (FMDV) on lateral flow device (LFD, penside test routinely used in the field for rapid immunodetection of FMDV), allowing its subsequent detection and typing by RT-PCR and recovery of live virus upon RNA transfection into permissive cells. After live FMDV collection onto LFD strip and soaking in 0.2% citric acid solution, the virus is totally inactivated. Viral RNA is still detectable by real-time RT-PCR following inactivation, and the virus strain can be characterized by sequencing of the VP1 coding region. In addition, live virus can be rescued by transfecting RNA extract from treated LFD into cells. This protocol should help promoting submission of FMD suspected samples to reference laboratories (by reducing the cost of sample shipping) and thus characterization of FMDV strains circulating in endemic regions.

Genetic Characterization of Serotypes A and Asia-1 Foot-and-mouth Disease Viruses in Balochistan, Pakistan, in 2011.

This study reports characterization of foot-and-mouth disease virus (FMDV) in samples collected from Balochistan, Pakistan. FMDV was detected by pan-FMDV real-time RT-PCR in 31 samples (epithelial and oral swabs) collected in 2011 from clinical suspect cases. Of these, 29 samples were serotyped by serotype-specific real-time RT-PCR assays and were confirmed by sequencing the VP1 coding region. Sixteen samples were found positive for serotype A and eight for serotype Asia-1, whereas five samples were found positive for both serotypes A and Asia-1. Two serotype A positive samples were found positive for two different strains of serotype A FMDV each. Phylogenetic analyses of serotype A FMDVs showed circulation of at least three different sublineages within the A-Iran05 lineage. These included two earlier reported sublineages, A-Iran05HER-10 and A-Iran05FAR-11 , and a new sublineage, designated here as A-Iran05BAL-11 . This shows that viruses belonging to the A-Iran05 lineage are continuously evolving in the region. Viruses belonging to the A-Iran05FAR-11 sublineage showed close identity with the viruses circulating in 2009 in Pakistan and Afghanistan. However, viruses belonging to the A-Iran05HER-10 detected in Balochistan, Pakistan, showed close identity with the viruses circulating in Kyrgyzstan, Iran and Kazakhstan in 2011 and 2012, showing that viruses responsible for outbreak in these countries have a common origin. Serotype Asia-1 FMDVs reported in this study all belonged to the earlier reported Group-VII (Sindh-08), which is currently a dominant strain in the West Eurasian region. Detection of two different serotypes of FMDV or/and two different strains of the same serotype in one animal/sample shows complexity in occurrence of FMD in the region.

Re-Emergence of Bluetongue Virus Serotype 8 in France, 2015.

At the end of August 2015, a ram located in central France (department of Allier) showed clinical signs suggestive of BTV (Bluetongue virus) infection. However, none of the other animals located in the herd showed any signs of the Bluetongue disease. Laboratory analyses identified the virus as BTV serotype 8. The viro and sero prevalence intraherd were 2.4% and 8.6% in sheep and 18.3% and 42.9% in cattle, respectively. Phylogenetic studies showed that the sequences of this strain are closely related to another BTV-8 strain that has circulated in France in 2006-2008. The origin of the outbreak is unclear but it may be assumed that the BTV-8 has probably circulated at very low prevalence (possibly in livestock or wildlife) since its first emergence in 2007-2008.

Establishment and validation of two duplex one-step real-time RT-PCR assays for diagnosis of foot-and-mouth disease.

Two duplex one-step TaqMan-based RT-PCR protocols for detection of foot-and-mouth disease virus (FMDV) were established and validated. Each RT-PCR test consists of a ready-to-use master mix for simultaneous detection of the well established 3D or IRES FMDV targets and incorporates the host ß-actin mRNA as an internal control target, in a single-tube assay. The two real-time RT-PCR 3D/ß-actin and IRES/ß-actin tests are highly sensitive and able to detect up to 7TCID50/ml of FMDV and 10 copies/1µl of viral RNA. In field epithelium samples, the diagnostic sensitivity was 100% (95% CI; 91-100%) for the 3D/ß-actin test and 97% (95% CI; 87-100%) for the IRES/ß-actin test. The diagnostic specificity was 100% (95% CI; 95-100%) for both RT-PCRs. In addition, the two protocols proved to be robust, showing inter-assay coefficients of variation ranging from 1.94% to 6.73% for the IRES target and from 2.33% to 5.42% for the 3D target for different RNA extractions and different RT-PCR conditions. The internally controlled one-step real-time RT-PCR protocols described in this study provide a rapid, effective and reliable method for the detection of FMDV and thus may improve the routine diagnosis for foot-and-mouth disease.

Psychosocial functioning in relation to symptomatic remission: A longitudinal study of first episode schizophrenia.

OBJECTIVES: The aims of the study were: (1) to evaluate longitudinally symptomatic remission in first-episode (FE) schizophrenia, (2) to describe symptoms, social functioning and quality of life (Qol) in relation to remission status, and (3) to determine the long-term outcome of schizophrenia and its early predictors. METHODS: Sixty-four patients were assessed 1 month after a first hospitalization (T1), 12 months (T2), 4-6 years (T3), and 7-11 years (T4) after T1. The patients were allocated to three remission groups according to their remission status over the whole observation period, e.g. stable remission (SR), unstable remission (UR) and non-remission (NR). The PANSS, Social Functioning Scale and WHOQoL were used to evaluate the patients' psychosocial functioning levels, symptomatic and functional remissions and satisfying QoL. A good outcome was defined as meeting, simultaneously, the criteria of symptomatic and functional remissions and satisfying QoL at T4, while failure to meet all of these criteria was defined as a poor outcome. RESULTS: Among them, 17.2% patients were in stable remission, 57.8% in unstable remission and 25.0% were unremitted at all time points. The SR group had lower levels of psychopathological symptoms and reported better social functioning and QoL than the NR group. During the follow-up, the symptoms increased, social functioning slightly improved and QoL did not change. At T4, 53% of the sample had a poor outcome, which was independently predicted by the longer duration of untreated psychosis and a lack of satisfying QoL at T1. CONCLUSIONS: Our results demonstrate that: (1) the long-term course in schizophrenia is heterogeneous and that three illness trajectories exist, (2) social functioning and QoL are only partially connected with symptomatic remission (3), the risk of a poor outcome may potentially be reduced by appropriate interventions at an early stage of the illness.

Factors contributing to cognitive deficits in elderly residents of rural areas.

OBJECTIVE: Due to the growing aging of societies an increasingly large group of people suffers from age-related impairment of cognitive functions and thus reducing the quality of life of the elderly. The purpose of the study was to evaluate the efficiency of cognitive functions in a group of aging residents of rural areas. PATIENTS AND METHODS: The inhabitants of a rural area were recruited and assessed: cognitive function as well as intellectual and physical activity, number of years of education, presence of diseases, using stimulants, diet, sources of living, marital status and family situation Subjects were divided into two groups: persons above 65 and older, constituting the studied group and persons between 40 and 64 years of age, constituting the control group. Both groups did not significantly differ in terms of sex or years of education. RESULTS: Statistically significant differences (p < 0.05) were found in the results of the tests concerning such functions as the sight recognition memory and spatial recognition memory, spatial operating memory both on the strategy level and on the level of committed errors. An analysis of the results obtained in the group of elderly people did not indicate any major differences between men and women as regards the analyzed cognitive functions, no statistically significant differences were found in cognitive testing depending on the number of years of education. The studied persons included in the physically active group scored better in the visual memory and learning tests. CONCLUSIONS: The conducted studies elucidated the dependence of the level of cognitive functions on age, a positive impact of physical activity on some cognitive functions, however we could not find differences between the efficiency of those functions and education, sex, presence of somatic diseases and activity of persons aged > 65.

Association of cognitive performance with the physical activity and body mass index in middle-aged and older rural inhabitants.

OBJECTIVE: Last few decades have witnessed rapid ageing of the population. The prevalence of mild cognitive impairment and dementia is significantly higher in people aged 60 and over than in younger. The aim of the study was the assessment of the putative associations between physical activity and cognitive functioning in elderly inhabitants of a rural area. PATIENTS AND METHODS: The participants underwent physical assessment, physical activity, cognitive functions, depression were assessed. RESULTS: Highly active participants had better results in tests assessing psychomotor speed and in measuring attention and its flexibility comparing to groups of medium and low physical activity. Active people made fewer errors in the test measuring visual memory and new learning, and had significantly better results in the spatial-recognition memory test. Persons with symptoms of depression performed worse in visuospatial memory and working memory tests. Associations between physical activity and cognitive test results as well as association between body mass index and blood pressure and cognitive performance were observed. CONCLUSIONS: The results of the study suggest an association between higher physical activity and better cognitive functioning in the field of working memory and visual and spatial-recognition, attention and attention flexibility. Symptoms of depression, hypertension and increased BMI may adversely affect cognitive performance in elderly.

Encephalomyocarditis virus 2A protein is required for viral pathogenesis and inhibition of apoptosis.

The encephalomyocarditis virus (EMCV), a Picornaviridae virus, has a wide host spectrum and can cause various diseases. EMCV virulence factors, however, are as yet ill defined. Here, we demonstrate that the EMCV 2A protein is essential for the pathogenesis of EMCV. Infection of mice with the B279/95 strain of EMCV resulted in acute fatal disease, while the clone C9, derived by serial in vitro passage of the B279/95 strain, was avirulent. C9 harbored a large deletion in the gene encoding the 2A protein. This deletion was incorporated into the cDNA of a pathogenic EMCV1.26 strain. The new virus, EMCV1.26Δ2A, was capable of replicating in vitro, albeit more slowly than EMCV1.26. Only mice inoculated with EMCV1.26 triggered death within a few days. Mice infected with EMCV1.26Δ2A did not exhibit clinical signs, and histopathological analyses showed no damage in the central nervous system, unlike EMCV1.26-infected mice. In vitro, EMCV1.26Δ2A presented a defect in viral particle release correlating with prolonged cell viability. Unlike EMCV1.26, which induced cytopathic cell death, EMCV1.26Δ2A induced apoptosis via caspase 3 activation. This strongly suggests that the 2A protein is required for inhibition of apoptosis during EMCV infection. All together, our data indicate that the EMCV 2A protein is important for the virus in counteracting host defenses, since Δ2A viruses were no longer pathogenic and were unable to inhibit apoptosis in vitro.

Quality of life and depression in schizophrenic patients.

PURPOSE: The aim of the study was to assess depressive symptoms, and to establish their influence on the subjective and objective quality of life (QOL) in schizophrenia patients. MATERIAL AND METHODS: Seventy four subjects: 46 male and 28 female, aged 24.7 +/- 6.7 years, were enrolled for the study. World Health Organization of Life Instrument-Bref (WHO-QOL-BREF), Social Functioning Scale (SFS) and Calgary Depression Scale for Schizophrenia (CDSS) were used. RESULTS: Severity of depressive symptoms showed moderate correlation with objective and strong correlation with subjective measures of QOL. CONCLUSIONS: Detection and appropriate treatment of depressive symptoms in schizophrenic patients may affect their functioning and perception of own health.

Biodegradable polyurethane cytocompatibility to fibroblasts and staphylococci.

Biodegradable polyurethanes have potential for use as implantable devices (orthopedic, maxillofacial, cardiovascular, wound dressing and plastic surgery) because of their controllable elasticity, and the possibility of changing their chemistry and structure. Studying bacterial and cell adhesion to polyurethanes helps to determine surface cytocompatibility and suitability for in vivo trials. Staphylococcus aureus, Staphylococcus epidermidis and hTERT human fibroblast cells were used to determine the cytocompatibility of experimental biodegradable polyurethanes (PUs) with different hydrophobic-to-hydrophilic (pho:phi) content ratios (100% pho, 70% pho, and 30% pho). Poly(L/DL-lactide) 70/30% (PLDL) and Thermanox were used as control surfaces. Surface characterization using noncontact profilometry, contact angles, and scanning electron microscopy (SEM) showed that the three PU surfaces, PLDL, and Thermanox have different properties. On the 100% PU and 30% PU surfaces, hTERT cells spread less in comparison to the 70% PU, PLDL, and Thermanox surfaces. The adsorption of fibronectin to the surfaces had no effect on the adhesion and spreading of hTERT cells when compared to the uncoated surfaces. The trend for S. aureus was the most adhered on the 70% PU and 30% PU, then Thermanox, followed by 100% PU and PLDL, respectively. The amount of S. epidermidis adhesion followed the trend of the most on 70% PU, then 100% PU, then 30% PU and PLDL, and the least on Thermanox. These results suggest that the 70% PU surface is cytocompatible to hTERT fibroblasts, while the 100% PU and 30% PU were not. All surfaces encouraged S. aureus and S. epidermidis colonization, particularly the 70% PU.

Burnout, stress and styles of coping among hospital nurses.

PURPOSE: The evaluation of professional burnout among hospital nurses and the analysis of correlations between burnout and a subjectively perceived stress and coping styles. MATERIAL AND METHODS: A study sample consisted of 227 nurses from general medical, neurological and psychiatric hospital wards. A set of 3 questionnaires was used, including Maslach Burnout Inventory (MBI), Coping Inventory for Stressful Situations (CISS) and Subjectively Perceived Stresss (SPS). RESULTS: Average and high level of burnout in the emotional exhaustion (EE), depersonalisation (D) and personal accomplishment (PA) was present at 71%, 39.8% and 77% of nurses respectively. A significantly higher level of burnout was noted in the subgroup of general medical nurses. The diferences involved the total MBI score and the results of the subscales EE and D (p < 0.01). A significant correlation has been found between the subjectively perceived stress and the level of burnout (r = 0.51, p < 0.01). Significant correlations has been found between MBI scores and CISS scores. Correlation between burnout and a task oriented coping was negative and correlation between burnout and emotion oriented coping was positive. CONCLUSIONS: The level of stress influences the professional burnout among nurses. There is a diversity in the level of burnout depending on the specialization at work, which is not accompanied by a similar diversity in the subjectively perceived stress. The correlation between burnout and a coping style is rather weak, but statistically significant.

Objective and subjective quality of life in schizophrenic patients after a first hospitalization.

PURPOSE: There is no single, universally accepted definition of quality of life (QOL). Both subjective and objective information is necessary to assess QOL. The aim of the study was to evaluate in cross-sectional and prospective manner objective and subjective quality of life in schizophrenic patients 1 month after hospitalization and in one year follow-up. MATERIAL AND METHODS: A study sample consisted of 86 schizophrenic subjects: 52 male and 34 female; age 25.5; +/-5.8 (range 17-47) and control group of matched 52 male and 34 female subjects were enrolled. Subjective QOL scale (WHOQOL-BREF), Social Functioning Scale (SFS) and structured questionnaire were used. Patients were evaluated 1 month (T1) and 13 months (T2) after a discharge from the hospital. RESULTS: In both T1 and T2 we found similar levels of SFS score and subjective measurement of QOL in patients, which were significantly lower than in healthy controls. CONCLUSIONS: This study showed that both objective and subjective quality of life are significantly decreased directly after hospitalization, and they are relatively stable in 1-year follow-up.

DNA released from ischemic and rejecting organs as an indicator of graft cellular damage.

Donor cellular debris contains fragments of nuclei with genetic material. The question arises whether the amount of donor graft released DNA accumulating in the recipient lymphoid tissues after transplantation could be a measure of donor organ damage caused by ischemia and preservation as well as rejection. We found that donor heart passenger cells do not contribute to the DNA disseminated in the recipient. All donor DNA was, then, derived from the damaged graft cells. Immediately and 1 day after transplantation, it was present in blood (plasma and cells) to accumulate later in the spleen. Higher values of donor DNA in the syngeneic than allogeneic combination, most evident on day 7, were presumably due to better perfusion of graft not undergoing rejection. Immunosuppression attenuated donor DNA release and accumulation in recipient tissues, nevertheless, relatively high concentrations could still be detected. Further studies are in progress on the usefulness of measuring DNA concentration for evaluation of the graft damage.

[Social functioning of patients with schizophrenia: a follow-up study]. / Funkcjonowanie spoleczne chorych na schizofrenie (badania katamnestyczne).

The subject of this study was evaluation of social functioning of schizophrenic patients after a period of ten to twenty years of follow-up. 70 patients (29 men and 41 women) selected from the initial group of 303 patients (181 men and 122 women), with the diagnosis of schizophrenia were studied. They had been admitted for the first time to the Hospital for Mental and Nervous Diseases in Gniezno in the years 1975-76. Examinations were carried in household environment (interviews with patients and their relatives), based on of a standardized questionnaire. In the examination, six realms of social activity were considered: professional occupation, home occupation, self independence, participation in family life, care for a child, social relations and interests. The level of social functioning in the examined group of 70 schizophrenic patients several years after the first psychiatric hospitalization has been found satisfactory in 57% patients. Women were functioning better, especially in the scope of participation in family life and fulfilment of home duties. Better level of social functioning was related to lower family history of mental illness, lower degree of invalidization, activity attitude towards the illness and positive self evaluation of the state of health, advantageous family situation, better fulfilment of social needs and lower family burden caused by the patient.