Scoring biosecurity in European conventional broiler production.

Good biosecurity procedures are crucial for healthy animal production. The aim of this study was to quantify the level of biosecurity on conventional broiler farms in Europe, following a standardized procedure, thereby trying to identify factors that are amenable to improvement. The current study used a risk-based weighted scoring system (biocheck.ugent ®) to assess the level of biosecurity on 399 conventional broiler farms in 5 EU member states. The scoring system consisted of 2 main categories, namely external and internal biosecurity, which had 8 and 3 subcategories, respectively. Biosecurity was quantified by converting the answers to 97 questions into a score from 0 to 100. The minimum score, "0," represents total absence of any biosecurity measure on the broiler farm, whereas the maximum score, "100," means full application of all investigated biosecurity measures. A possible correlation between biosecurity and farm characteristics was investigated by multivariate linear regression analysis. The participating broiler farms scored better for internal biosecurity (mean score of 76.6) than for external biosecurity (mean 68.4). There was variation between the mean biosecurity scores for the different member states, ranging from 59.8 to 78.0 for external biosecurity and from 63.0 to 85.6 for internal biosecurity. Within the category of external biosecurity, the subcategory related to "infrastructure and vectors" had the highest mean score (82.4), while the subcategory with the lowest score related to biosecurity procedures for "visitors and staff" (mean 51.5). Within the category of internal biosecurity, the subcategory "disease management" had the highest mean score (65.8). In the multivariate regression model a significant negative correlation was found between internal biosecurity and the number of employees and farm size. These findings indicate that there is a lot of variation for external and internal biosecurity on the participating broiler farms, suggesting that improvements are possible. Since the subcategory "visitors and staff" scored the lowest, better education of broiler farmers and their staff may help to improve overall biosecurity on broiler farms in Europe.

Gut microbial metabolism of polyphenols from black tea and red wine/grape juice is source-specific and colon-region dependent.

The colonic microbial degradation of a polyphenol-rich black tea extract (BTE) and red wine/grape juice extract (RWGE) was compared in a five-stage in vitro gastrointestinal model (TWINSHIME). Microbial metabolism of BTE and RWGE polyphenols in the TWINSHIME was studied subsequently in single- and continuous-dose experiments. A combination of liquid or gas chromatography with mass spectrometry (LC-MS or GC-MS) and NMR-based metabolic profiling was used to measure selected parent polyphenols, their microbial degradation into phenolic acids, and the production of short-chain fatty acids (SCFAs) in different colon compartments. Acetate production was increased by continuous feeding of BTE but not RWGE. During RWGE feeding, gallic acid and 4-hydroxyphenylpropionic acid remained elevated throughout the colon, while during BTE feeding, they were consumed in the distal colon, while 3-phenylpropionic acid was strongly produced. Gut microbial production of phenolics and SCFAs is dependent on colon location and polyphenol source, which may influence potential health benefits.

Restriction of trophic factors and nutrients induces PARKIN expression.

Parkinson's disease (PD) is the most frequent neurodegenerative movement disorder and manifests at old age. While many details of its pathogenesis remain to be elucidated, in particular the protein and mitochondrial quality control during stress responses have been implicated in monogenic PD variants. Especially the mitochondrial kinase PINK1 and the ubiquitin ligase PARKIN are known to cooperate in autophagy after mitochondrial damage. As autophagy is also induced by loss of trophic signaling and PINK1 gene expression is modulated after deprivation of cytokines, we analyzed to what extent trophic signals and starvation stress regulate PINK1 and PARKIN expression. Time course experiments with serum deprivation and nutrient starvation of human SH-SY5Y neuroblastoma cells and primary mouse neurons demonstrated phasic induction of PINK1 transcript up to twofold and PARKIN transcript levels up to sixfold. The corresponding threefold starvation induction of PARKIN protein was limited by its translocation to lysosomes. Analysis of primary mouse cells from PINK1-knockout mice indicated that PARKIN induction and lysosomal translocation occurred independent of PINK1. Suppression of the PI3K-Akt-mTOR signaling by pharmacological agents modulated PARKIN expression accordingly. In conclusion, this expression survey demonstrates that PARKIN and PINK1 are coregulated during starvation and suggest a role of both PD genes in response to trophic signals and starvation stress.

[Psychiatric sequelae of severe burn injuries: emotional distress and resources of occupationally versus non occupationally insured patients 1 year after burn injury]. / Psychische Folgen schwerer Brandverletzungen: Belastungen und Ressourcen berufsgenossenschaftlich vs. nicht berufsgenossenschaftlich Versicherter ein Jahr nach der Brandverletzung.

Severe burn injuries are traumatic events and can have serious impact on all areas of life frequently causing high emotional distress. In a multicentre study resources and emotional distress of patients with serious burn injuries were assessed during the first hospitalization and at 6 and 12 months follow-up. Patients with severe burn injuries after accidents in a private environment (NBG patients) and patients after occupational accidents covered by the German Social Accident Insurance (BG patients) were compared. All patients reported marked emotional impairment, particularly during the hospitalization. At follow-up a reduction of emotional distress was detected. Nearly half of the patients received a diagnosis of one or more mental disorders according to DSM-IV criteria. When treating patients with burns, special attention should be given to their mental health. They should be offered psychological support to cope with the aftermath of the accident, especially after discharge from hospital when returning to their normal surroundings.

N(2)-BET specific surface area of bentonites.

The specific surface areas (SSA(N2BET)) of 36 different bentonites had larger values for Ca(2+)/Mg(2+) bentonites than for Na(+) bentonites. This trend could not be explained by the different d(001) values nor by the different microstructures. The investigation of Cu-triene-exchanged smectites, which on drying at 105 degrees C still had a d(001) value accounting for approximately 13A, proved that the SSA(N2BET) of low-charged smectites increased more than that of high-charged smectites. This could be explained by: (i) more space between the permanent charge sites in the case of low-charged smectites and (ii) the fact that the layers of Cu-triene smectites do not collapse at 105 degrees C. In contrast the SSA(N2BET) of Ca(2+)-exchanged bentonites could not be related to the layer charge density (LCD) as in the case of the Cu-triene-exchanged bentonites which is probably due to the varying number of collapsed layers. In conclusion, the SSA(N2BET) of bentonites which is known to be largely variable is probably determined by microporosity resulting from the quasi-crystalline overlap region and accessible areas of the interlayer. The number of layers per stack and the microstructure are supposed to play a subordinate role. The larger SSA(N2BET) of Ca/Mg bentonites compared to Na bentonites probably can be explained by the larger space between the charges in the case of the presence of divalent cations.

Enhanced vulnerability of PARK6 patient skin fibroblasts to apoptosis induced by proteasomal stress.

Proteasomal dysfunction and apoptosis are major hallmarks in the pathophysiology of Parkinson's disease (PD). PARK6 which is caused by mutations in the mitochondrial protein kinase PINK1 is a rare autosomal-recessively inherited disorder mimicking the clinical picture of PD. To investigate the cytoprotective physiological function of PINK1, we used primary fibroblasts from three patients homozygous for G309D-PINK1 as well as SHEP neuroblastoma cells stably overexpressing GFP-tagged wild type (wt) PINK1. Here we demonstrate that overexpression of wt PINK1 inhibits activation of Bax and release of cytochrome c, thereby diminishing caspase 9 processing and effector caspase activity after induction of proteasomal stress with the proteasome inhibitor (PI) MG132 in SHEP cells. Conversely, effector caspase activation induced by PIs, but not by the unrelated apoptotic stimulus staurosporine was potently enhanced in primary fibroblasts from homozygous PARK6 patients in comparison to those of heterozygous carriers or unaffected siblings. SHEP cells overexpressing wt PINK1 showed an elevated expression of the cytoprotective gene parkin, whereas PARK6 fibroblasts displayed significantly decreased expression of parkin in comparison to wild type control cells. Interestingly, overexpressed GFP-PINK1 was exclusively localized in the mitochondria of SHEP cells, but was redistributed to the cytoplasm under conditions of proteasomal stress. Our data indicate that PINK1 plays an important and specific physiological role in protecting cells from proteasomal stress, and suggest that PINK1 might exert its cytoprotective effects upstream of mitochondria engagement.

[Religiosity and mood in the last week of life: an explorative approach based on after-death proxy-interviews]. / Religiositeit en stemming in de laatste levensweek; een exploratieve benadering aan de hand van nabestaandeninterviews.

The current contribution focuses on the role of religiousness on aspects of mood in the last week of life. After-death interviews with proxy respondents of deceased participants of the Longitudinal Aging Study Amsterdam provided information on depressive mood and anxiety in the last week of life, as well as on a sense of peace with the approaching end of life. Furthermore, the proxy respondents were asked about serious physical symptoms in the last week of life, cognitive decline, salience of religion, and whether the deceased respondent had talked about religion. Other characteristics were derived from the last interviews with the respondents when still alive: depressive symptoms, religious affiliation, church-attendance, orthodoxy, salience of religion, and cosmic transcendence. None of the characteristics of religiousness were significantly associated with depressive mood or anxiety as estimated by the proxy-respondent. A sense of peace, however, was predicted by higher church-attendance, Protestant affiliation (as compared to no affiliation), and the proxy's estimate of the salience of religion. It is concluded that religiousness does not affect depressive mood or anxiety in the last week of life in the current sample. Possibly, religiousness supports a sense of peace, which may be a more existential facet of mood, and which is discussed as relevant in the last phase of life and in palliative care.

Profile of the malnourished patient.

OBJECTIVE: To characterize malnutrition in a nonspecific group of newly admitted hospital patients. DESIGN: A prospective, descriptive study aiming to identify typical symptoms of malnutrition in a heterogeneous population of newly admitted patients to the wards of internal medicine. SETTING: The wards of internal medicine of the VU University Medical Center. SUBJECTS: A total of 106 patients were included in the study, 70 patients underwent the full interview. Next to nutritional status, the sociodemographics, underlying disease, estimated care complexity, care situation before admission, journey through the care system, nutritional intervention and nutritional follow-up after discharge were described for each patient. RESULTS: Of 70 patients 24 (34%) were malnourished. Malnourished patients suffered two chronic diseases vs one for well-nourished patients (P = 0.05). They also had a higher estimated care complexity (P = 0.035) and a trend towards longer length of hospital stay (P = 0.09). Malnourished patients did not differ from well-nourished patients in age, sex, partner status and care received at home. In all, 54% of the malnourished patients were identified correctly by the medical staff. The reasons for admission to the hospital were diverse in only four out of 24 patients malnutrition was the primary reason for admission. Discharge letters to the general practitioner (GP) contained only fragmentary information about the patients' nutritional status. At 3 months after discharge, most of the GPs were scarcely aware of any nutritional problems of their patients. CONCLUSIONS: Malnutrition is difficult to recognize in a nonspecific hospital population. Patients do not present with unique symptoms indicating malnutrition. To be able to correctly identify all malnourished patients, screening of the nutritional status of all newly admitted patients seems to be necessary.

The end of life: informal care for dying older people and its relationship to place of death.

OBJECTIVE: This study examined the features of informal end-of-life care of older people living in the community and the association between informal care characteristics and dying at home. METHODS: Retrospective data were obtained from interviews and self-administered questionnaires of 56 persons who had been primary caregivers of older relatives in the last three months of their lives. RESULTS: Results showed that informal caregivers of terminally ill older people living in the community provided a considerable amount of personal, household, and management care. Secondary informal caregivers and formal caregivers assisted resident primary caregivers less often than nonresident primary caregivers. Primary caregivers who felt less burdened, who gave personal care more intensively, and/or who were assisted by secondary caregivers, were more likely to provide informal end-of-life care at home until the time of death. CONCLUSIONS: Our study showed that informal care at the end of life of older people living in the community is complex, since the care required is considerable and highly varied, and involves assistance from secondary informal caregivers, formal home caregivers as well as institutional care. Burden of informal care is one of the most important factors associated with home death. More attention is needed to help ease the burden on informal caregivers, specifically with regard to resident caregivers and spouses. Since these resident caregivers were disadvantaged in several respects (i.e., health, income, assistance from other carers) compared to nonresident caregivers, interventions by formal caregivers should also be directed towards these persons, enabling them to bear the burden of end-of-life care.

Proxy reporting in after-death interviews: the use of proxy respondents in retrospective assessment of chronic diseases and symptom burden in the terminal phase of life.

This study evaluates the quality of data obtained from after-death interviews with significant others of deceased older persons regarding the prevalence of chronic diseases and symptoms in the terminal phase of life. These data are compared with reports from physicians and earlier self-reports from the deceased person. There were significant increases in nonresponse and nonavailability of significant others for decedents who had been divorced or had never been married, thus introducing some selection bias. At the level of the total sample, significant others seem to give accurate information about the prevalence of chronic diseases when compared with self-reports and reports from physicians. At the level of the individual sample member, after-death interviews with significant others provide valid information for the assessment of the prevalence of malignant neoplasms, diabetes mellitus, chronic obstructive pulmonary disease and cerebrovascular disease, but not for osteo- and rheumatoid arthritis and artherosclerotic disease. At the level of the total sample, the prevalence of symptoms assessed by significant others did not differ greatly from the assessment made by physicians. However, at the level of the individual sample member, the validity of symptom assessment by significant others could not be supported by data obtained from the physicians. With regard to the type of significant others interviewed, children reported more symptoms than partners. The use of significant others in after-death interviews can be a valid method with regard to the assessment of chronic diseases and symptoms on a group level. On an individual level this can be concluded only for chronic diseases with clearly observable consequences.

Functional redundancy of the zinc fingers of A20 for inhibition of NF-kappaB activation and protein-protein interactions.

The tumor necrosis factor (TNF) inducible protein A20 is a potent inhibitor of nuclear factor-kappaB (IkappaB)-mediated gene expression in response to TNF and several other stimuli. The C-terminal domain of A20 is characterized by seven zinc finger structures. Here, we show that a minimum of four zinc fingers is required to inhibit TNF-induced nuclear factor-kappaB (NF-kappaB) activation to a level that is comparable to that obtained with the wild-type A20 protein. However, there was no strict requirement for a particular zinc finger structure, since a mutant A20 protein containing only the first four zinc fingers was as potent as a mutant protein containing only the last four zinc fingers. A similar functional redundancy of the A20 zinc fingers was also observed for binding of A20 to a number of other proteins, including two novel NF-kappaB inhibitory proteins (ABIN-1, ABIN-2), A20 itself, the anti-apoptotic protein TXBP151, and a regulatory component of the IkappaB kinase complex, IKKgamma. Moreover, we demonstrate that complete loss of binding of any of these proteins correlates with complete loss of A20's ability to inhibit TNF-induced NF-kappaB activation. However, binding of IKKgamma as such is not sufficient for inhibition of NF-kappaB dependent gene expression in response to TNF.

A dominant negative mutation in Saccharomyces cerevisiae methionine aminopeptidase-1 affects catalysis and interferes with the function of methionine aminopeptidase-2.

Methionine aminopeptidase (MetAP) enzymes require the metal ion cobalt, but little is known about the role of cobalt in the structural stability or catalysis of these enzymes. In Escherichia coli MetAP, for which a crystal structure is available, the five amino acid residues liganding the two cobalt ions are Asp97, Asp108, His171, Glu204, and Glu235. These five amino acids are conserved in all MetAPs sequenced to date. The C-terminal domain of the yeast Saccharomyces cerevisiae MetAP1 is 41% identical to E. coli MetAP and contains these cobalt coordinating residues. Using site-directed mutagenesis on the gene coding for yeast MetAP1, we replaced Asp219 (corresponding to Asp97 in E. coli MetAP) with Asn. The yeast D219N mutant enzyme has 10(3)-fold lower catalytic activity and a different substrate specificity when compared to wild-type yeast MetAP1. These results indicate that the side-chain of Asp219 is important for catalysis. Expression of D219N-MetAP1 in yeast causes a slow-growth phenotype and interferes with wild-type MetAP1 in a dominant manner. Expression of D219N-MetAP1 also affects the function of S. cerevisiae MetAP2.

Mutational analysis of a patient with mucopolysaccharidosis type VII, and identification of pseudogenes.

PCR of cDNA produced from patient fibroblasts allowed us to determine the paternal mutation in the first patient reported with beta-glucuronidase-deficiency mucopolysaccharidosis type VII (MPS VII). The G-->T transversion 1,881 bp downstream of the ATG translation initiation codon destroys an MboII restriction site and converts Trp627 to Cys (W627C). Digestion of genomic DNA PCR fragments with MboII indicated that the patient and the father were heterozygous for this missense mutation in exon 12. Failure to find cDNAs from patient RNA which did not contain this mutation suggested that the maternal mutation leads to greatly reduced synthesis or reduced stability of mRNA from the mutant allele. In order to identify the maternal mutation, it was necessary to analyze genomic sequences. This approach was complicated by the finding of multiple unprocessed pseudogenes and/or closely related genes. Using PCR with a panel of human/rodent hybrid cell lines, we found that these pseudogenes were present over chromosomes 5-7, 20, and 22 and the Y chromosome. Conditions were defined which allowed us to amplify and characterize genomic sequences for the true beta-glucuronidase gene despite this background of related sequences. The patient proved to be heterozygous for a second mutation, in which a C-->T transition introduces a termination codon (R356STOP) in exon 7. The mother was also heterozygous for this mutation. Expression of a cDNA containing the maternal mutation produced no enzyme activity, as expected. Expression of the paternal mutation in COS-7 cells produced a surprisingly high (65% of control) level of activity. However, activity was 13% of control in transiently transfected murine MPS VII cells. The level of activity of this mutant allele appears to correlate with the level of overexpression, suggesting that high concentrations of mutant monomers can drive the folding and tetramerization of mutant enzyme to produce an active and stable enzyme.