Temporal regulation of murine cytomegalovirus transcription and mapping of viral RNA synthesized at immediate early times after infection

The replication of murine cytomegalovirus strain Smith in murine embryonic fibroblasts was investigated at immediate early, early, and late times after infection. Cloned subgenomic HindIII fragments of murine cytomegalovirus DNA served to define the regions of transcription. At immediate early times viral RNA classes ranging in size from 5.1 to 1.05 kilobases (kb) were transcribed mainly from the fragments HindIII-K and -L, whereas low levels of transcription were detected from the two termini HindIII-E and HindIII-N. A characteristic pattern of proteins could be translated from immediate early RNA in vitro. At early and late times after infection transcription from all HindIII fragments occurred, but different patterns of transcripts and proteins could be identified. Inhibitors of DNA synthesis induced differences in the late transcription pattern, located in the HindIII-F fragment. The coding region for abundant immediate early transcription could be located at between 0.769 and 0.817 map units. A plasmic clone containing the main part (0.769 to 0.815 map units) of this region was constructed. This region coded for six polyadenylated immediate early RNA species of 5.1, 2.75, 2.0, 1.75, 1.65, and 1.05 kb in size. Only the 1.75-kb RNA originated entirely from the HindIII-L fragment. The 5.1- and 2.75-kb RNA species were encoded by both the HindIII-L and HindIII-K fragments, and the 2.0-, 1.65-, and 1.05-kb RNA species were entirely transcribed within HindIII-K

Location, transcripts, and translation products

Cloned genomic fragments from the region (0.769 to 0.818 map units) coding for immediate-early (IE) transcripts of murine cytomegalovirus (MCMV) were used to analyze the physical organization of this region, the direction of transcription, and the proteins synthesized in vitro. Three IE transcription units could be identified. From IE coding region 1 (ie1; 0.781 to 0.796 map units) a dominant 2.75-kilobase (kb) RNA was transcribed from right to left on the prototype arrangement of the MCMV genome which directed the synthesis of an 89,000-molecular-weight polypeptide (89K polypeptide), the major IE protein. This phosphoprotein (pp89) has been shown to be active in the regulation of transcription. Upstream of ie1 and separated by the MCMV enhancer sequence was a second IE coding region, ie2, which was mapped at 0.803 to 0.817 map units. From ie2 a 1.75-kb RNA of moderate abundance was transcribed in the direction opposite to that of the ie1 RNA. After hybrid selection of the ie2 transcript, a 43,000-molecular-weight translation product was detected. A third coding region, ie3, was located directly downstream of ie1 (0.773 to 0.781 map units). The series of RNAs with low abundance, terminating in ie3, probably used the ie1 transcription start site and ranged from 1.0 to 5.1 kb in size. The 5.1-kb RNA apparently represents the nonspliced transcript from both coding regions ie1 and ie3. A 15K polypeptide was translated in vitro from RNA that was hybrid selected by ie3 sequences. Immunoprecipitation with monoclonal antibody revealed that 31K to 67K polypeptides were related to pp89. Some of these proteins were translated from RNAs that were smaller than 2.75 kb. Polypeptides related to pp89 were also synthesized in vivo. Because polypeptides unrelated to pp89 that were translated from RNA that was selected by ie2 and ie3 sequences were not immunoprecipitated by murine antisera, we assumed that the amount of these proteins synthesized in vivo during infection was probably very lo

Die landwirtschaftliche Baukultur nachhaltiger gestalten und die kommunikative Wirkung der Bausubstanz erkennen

Gebäude sind nicht nur als Funktionseinheiten zu betrachten, sondern auch als Kommunikationsmedium. Die bauliche Gestaltung eines jeweiligen Gebäudes lässt viele Rückschlüsse bezüglich der Einstellungen und Werthaltungen der Bauherrin oder des Bauherrn zu. Und so geben auch Schweineställe und Feldscheunen Informationen über das Wertegefüge des jeweiligen landwirtschaftlichen Unternehmens preis. Die finanziellen Spielräume der Agrarbetriebe sind oftmals nicht groß bei baulichen Investitionen. Unreflektiert sollte die agrarische Baukultur deswegen allerdings nicht bleiben. Die folgenden Ausführungen zeigen auf, dass ein landwirtschaftlicher Betrieb durch eine reflektierte Baukultur einen Zugewinn an Authentizität in der öffentlichen Wahrnehmung erfahren kann

Veränderungen der Konsumintensität von Öko-Lebensmitteln in Haushalten mit Kindern

Considering results of consumer surveys the question arises that on the one hand the health of children is a major motive for organic food consumption in households with children and why on the other hand consumption intensity decreases as children get older. The contribution presents results of a qualitative study investigating changes in families’ organic food consumption and their reasons from the subjective perspective of consumers. The findings are based on information gained by problem centred interviews with family members responsible for food purchases; data analysis was done by means of Grounded Theory. It was found that children can be reasons for an increase of organic food consumption by indirectly influencing parents’ willingness to purchase organic food in the time of pregnancy and the transition to a complementary diet for babies. In contrast to that children also can directly affect a decrease of organic food consumption in their families by actively rejecting particular organic food products

The 89,000-Mr murine cytomegalovirus immediate-early protein activates gene transcription

To study trans-activation of gene expression by murine cytomegalovirus (MCMV) immediate-early (IE) proteins, the IE coding region 1 (ie1), which encodes the 89,000-Mr IE phosphoprotein (pp89), was stably introduced into L cells. A cell line was selected and characterized that efficiently expressed the authentic viral protein. The pp89 that was constitutively expressed in L cells stimulated the expression of transfected recombinant constructs containing the bacterial chloramphenicol acetyltransferase (CAT) gene under the control of viral promoters. The regulatory function of the ie1 product was confirmed by transient expression assays in which MCMV IE genes were cotransfected into L cells together with recombinant constructs of the CAT gene. For CAT activation by the ie1 product, a promoter region was required, but there was no preferential activation of a herpes simplex virus type 1 delayed-early promoter. All plasmid constructs that contained the intact coding sequences for pp89 induced gene expression in trans. The MCMV enhancer region was not essential for the expression of a functional IE gene product, and testing of the cis-regulatory activity of the MCMV enhancer revealed a low activity in L cells. Another region transcribed at IE times of infection, IE coding region 2, was unable to induce CAT expression and also did not augment the functional activity of ie1 after cotransfection

„Hauptsache, es schmeckt“ - der Einfluss von Jugendlichen auf den Öko-Lebensmittelkonsum in Familien

This contribution presents results of a qualitative study investigating juveniles’ influence in family decision-making with regard to organic food. According to recent research it can be assumed that expenditures for organic food in families with children decline as children grow older. For organic food marketing this raises the questions if, why and how juveniles influence family decision-making and what organic products are prefered or rejected? Semi-structured interviews were conducted with juveniles aged 13 to 18 years. The data was analysed using qualitative content analysis. Most juveniles proved to be open-minded towards organic food and they supported its consumption at home. Juveniles’ main criticism encompassed the taste of some organic product groups such as sweets, crisps, spreads and pizzas. If juveniles are not satisfied with the taste of an organic product they use different strategies to influence family purchase decisions. The main aim of these strategies is often to buy a conventional product. Therefore, organic suppliers should invest more money into researching juveniles’ specific taste preferences and should develop products which meet the expectations of juveniles better with respect to taste, but also regarding product appearance and packaging. Via advertising, product images should be optimised to comply with the actual lifestyle of the target group. Ethical aspects should also be included into communication strategies, as juveniles have a special sense for social justice and animal welfare

Medication knowledge of patients hospitalized for heart failure at admission and after discharge

Background: A substantial aspect of health literacy is the knowledge of prescribed medication. In chronic heart failure, incomplete intake of prescribed drugs (medication non-adherence) is inversely associated with clinical prognosis. Therefore, we assessed medication knowledge in a cohort of patients with decompensated heart failure at hospital admission and after discharge in a prospective, cross-sectional study. Methods: One hundred and eleven patients presenting at the emergency department with acute decompensated heart failure were included (mean age 78.4±9.2, 59% men) in the study. Patients’ medication knowledge was assessed during individual interviews at baseline, course of hospitalization, and 3 months after discharge. Individual responses were compared with the medical records of the referring general practitioner. Results: Median N-terminal prohormone of brain natriuretic peptide plasma concentration in the overall population at baseline was 4,208 pg/mL (2,023–7,101 pg/mL [interquartile range]), 20 patients died between the second and third interview. The number of prescribed drugs increased from 8±3 at baseline to 9±3 after 3 months. The majority of patients did not know the correct number of their drugs. Medication knowledge decreased continuously from baseline to the third interview. At baseline, 37% (n=41) of patients stated the correct number of drugs to be taken, whereas only 18% (n=16) knew the correct number 3 months after discharge (P=0.008). Knowledge was inversely related to N-terminal prohormone of brain natriuretic peptide levels. Conclusion: Medication knowledge of patients with acute decompensated heart failure is poor. Despite care in a university hospital, patients’ individual medication knowledge decreased after discharge. The study reveals an urgent need for better strategies to improve and promote the knowledge of prescribed medication in these very high-risk patients

Self‐compassion and emotion regulation difficulties in obsessive–compulsive disorder

Lack of self‐compassion and deficits in emotion regulation are associated with various psychopathological symptoms and may play a role in the development and maintenance of obsessive–compulsive disorder (OCD). However, further empirical research is still needed to better understand these constructs in the context of this disorder. The present study investigated the relation between self‐compassion, emotion regulation difficulties, obsessive beliefs, and obsessive–compulsive symptom severity in 90 patients with OCD using self‐report questionnaires. Symptom severity and obsessive beliefs were negatively correlated to self‐compassion and positively associated with emotion regulation difficulties. Additionally, self‐compassion showed a negative relation to emotion regulation difficulties. Emotion regulation difficulties—but not self‐compassion—predicted symptom severity when controlling for obsessive beliefs and depression in a hierarchical regression analysis. Further analyses showed that emotion regulation deficits mediated the relationship between self‐compassion and OCD symptom severity. Our results provide preliminary evidence that targeting self‐compassion and putting more emphasis on emotion regulation deficits might be promising treatment approaches for patients with OCD. Future studies could investigate which specific interventions that directly address these variables improve treatment outcome