IL-17F co-expression improves cell growth characteristics and enhances recombinant protein production during CHO cell line engineering.

The generation of a high productivity cell line is a critical step in the production of a therapeutic protein. Many innovative engineering strategies have been devised in order to maximize the expression rate of production cells for increased process efficiency. Less effort has focused on improvements to the cell line generation process, which is typically long and laborious when using mammalian cells. Based on unexpected findings when generating stable CHO cell lines expressing human IL-17F, we studied the benefit of expressing this protein during the establishment of production cell lines. We demonstrate that IL-17F expression enhances the rate of selection and overall number of selected cell lines as well as their transgene expression levels. We also show that this benefit is observed with different parental CHO cell lines and selection systems. Furthermore, IL-17F expression improves the efficiency of cell line subcloning processes. IL-17F can therefore be exploited in a standard manufacturing process to obtain higher productivity clones in a reduced time frame.

Regulierungen an der Schnittstelle zwischen den Ressourcen Wald und Klima : Einflussfaktoren auf die Inwertsetzung der CO₂-Senkenleistung des Waldes

Les forêts dont le volume sur pied augmente contribuent à atténuer les changements climatiques et constituent un puits de carbone. En Suisse, seul très peu d'entreprises forestières valorisent le service de puits de carbone. Ce travail vise à chercher les raisons expliquant cette réticence. Les facteurs politiques et juridiques influençant les décisions des entreprises forestières sont analysées à l'aide du modèle des régimes institutionnels de ressource. Les résultats obtenus sont vérifiés et complétés à l'aide d'informations collectées lors d'interviews d'experts menées auprès d'entreprises forestières et décideurs politiques. Cette étude identifie des facteurs favorisant ou entravant la valorisation du service de puits de carbone des forêts et propose des options d'action.

Hypertension and intra-operative incidents : a multicentre study of 125000 surgical procedures in Swiss hospitals

Rapport de synthèse : Bien que les complications sévères de l'anesthésie soient actuellement rares, des informations contradictoires existent à propos du rôle et de l'importance de l'hypertension artérielle chronique sur la survenue de complications en cours d'anesthésie. En raison de la prévalence élevée de l'hypertension artérielle dans la population et du grand nombre d'anesthésies effectuées, il est important de clarifier cette relation. Le but de l'étude était d'évaluer si les personnes hypertendues étaient à risque accru de présenter des complications lors d'anesthésies à partir de données collectées de routine lors d'anesthésies usuelles réalisées en Suisse. Nous avons utilisé les données figurant dans le registre ADS (Anesthésie Données Suisse) correspondant à des anesthésies, générales ou locorégionales, réalisées pour chirurgie élective chez des adultes, entre 2000 et 2004 dans 24 hôpitaux suisses. L'attention était portée principalement sur les incidents cardio-vasculaires, mais les autres incidents relevés de routine ont aussi été évalués. La présence d'une hypertension artérielle chronique était définie par la présence d'un traitement antihypertenseur ou par l'anamnèse d'une hypertension artérielle, combinée à la mesure d'une pression artérielle élevée (systolique >160 mm Hg ou diastolique >100 mm Hg) lors de l'examen préopératoire de l'anesthésiste. Les incidents relevés en cours d'anesthésie ont été définis a priori et sont enregistrés de routine sur la feuille d'anesthésie et reportés dans une base de données centralisée. En raison de la structure des données, des analyses hiérarchiques ont été effectuées incluant des variables individuelles (niveau 1), liées aux groupes d'interventions chirurgicales (niveau 2) et à l'hôpital (niveau 3). Parmi les 124 939 interventions, 27 881 (22%) concernaient des patients connus pour une hypertension artérielle chronique. Au moins un incident est survenu dans 16,8% des interventions (95% Cl 16,6 -17,0%). Chez 7 549 patients, au moins un incident cardio- vasculaire est survenu, soit dans 6% des anesthésies (95% Cl 5.9-6.2%). Le rapport des cotes (odds ratio) moyen ajusté pour les incidents cardio-vasculaires chez les patients hypertendus était de 1.38 (95% Cl 1.27-1.49), indiquant une augmentation du risque chez les patients hypertendus. Cependant, l'hypertension n'était pas liée à un risque augmenté de survenue d'un autre incident. Les rapports de cotes ajustés de la survenue d'une complication cardiovasculaire en présence d'une hypertension artérielle variaient selon les hôpitaux entre 0.41 et 2.25. Ainsi, cette étude confirme la présence d'un risque accru de survenue d'une complication cardiovasculaire chez un patient hypertendu lors d'une anesthésie pour chirurgie élective. Il s'agissait le plus souvent d'une arythmie ou d'une perturbation hémodynamique. Cette augmentation du risque proche de 40% a aussi été trouvée dans une revue systématique avec méta-analyse. L'hétérogénéité des institutions -qui persiste même en tenant compte des ajustements pour le type d'intervention chirurgicale et des variables individuelles (case-mix) - suggère des différences de pratique de l'anesthésie selon l'hôpital.

Peri-operative management of antiplatelet therapy in patients with coronary artery disease. Joint position paper by members of the working group on Perioperative Haemostasis of the Society on Thrombosis and Haemostasis Research (GTH), the working group on Perioperative Coagulation of the Austrian Society for Anesthesiology, Resuscitation and Intensive Care (ÖGARI) and the Working Group Thrombosis of the European Society for Cardiology (ESC).

An increasing number of patients suffering from cardiovascular disease, especially coronary artery disease (CAD), are treated with aspirin and/or clopidogrel for the prevention of major adverse events. Unfortunately, there are no specific, widely accepted recommendations for the perioperative management of patients receiving antiplatelet therapy. Therefore, members of the Perioperative Haemostasis Group of the Society on Thrombosis and Haemostasis Research (GTH), the Perioperative Coagulation Group of the Austrian Society for Anesthesiology, Reanimation and Intensive Care (ÖGARI) and the Working Group Thrombosis of the European Society of Cardiology (ESC) have created this consensus position paper to provide clear recommendations on the perioperative use of anti-platelet agents (specifically with semi-urgent and urgent surgery), strongly supporting a multidisciplinary approach to optimize the treatment of individual patients with coronary artery disease who need major cardiac and non-cardiac surgery. With planned surgery, drug eluting stents (DES) should not be used unless surgery can be delayed for ≥12 months after DES implantation. If surgery cannot be delayed, surgical revascularisation, bare-metal stents or pure balloon angioplasty should be considered. During ongoing antiplatelet therapy, elective surgery should be delayed for the recommended duration of treatment. In patients with semi-urgent surgery, the decision to prematurely stop one or both antiplatelet agents (at least 5 days pre-operatively) has to be taken after multidisciplinary consultation, evaluating the individual thrombotic and bleeding risk. Urgently needed surgery has to take place under full antiplatelet therapy despite the increased bleeding risk. A multidisciplinary approach for optimal antithrombotic and haemostatic patient management is thus mandatory.

The co-evolution of social institutions, demography, and large-scale human cooperation.

Human cooperation is typically coordinated by institutions, which determine the outcome structure of the social interactions individuals engage in. Explaining the Neolithic transition from small- to large-scale societies involves understanding how these institutions co-evolve with demography. We study this using a demographically explicit model of institution formation in a patch-structured population. Each patch supports both social and asocial niches. Social individuals create an institution, at a cost to themselves, by negotiating how much of the costly public good provided by cooperators is invested into sanctioning defectors. The remainder of their public good is invested in technology that increases carrying capacity, such as irrigation systems. We show that social individuals can invade a population of asocials, and form institutions that support high levels of cooperation. We then demonstrate conditions where the co-evolution of cooperation, institutions, and demographic carrying capacity creates a transition from small- to large-scale social groups.

The co-evolution of culturally inherited altruistic helping and cultural transmission under random group formation.

Limited migration results in kin selective pressure on helping behaviors under a wide range of ecological, demographic and life-history situations. However, such genetically determined altruistic helping can evolve only when migration is not too strong and group size is not too large. Cultural inheritance of helping behaviors may allow altruistic helping to evolve in groups of larger size because cultural transmission has the potential to markedly decrease the variance within groups and augment the variance between groups. Here, we study the co-evolution of culturally inherited altruistic helping behaviors and two alternative cultural transmission rules for such behaviors. We find that conformist transmission, where individuals within groups tend to copy prevalent cultural variants (e.g., beliefs or values), has a strong adverse effect on the evolution of culturally inherited helping traits. This finding is at variance with the commonly held view that conformist transmission is a crucial factor favoring the evolution of altruistic helping in humans. By contrast, we find that under one-to-many transmission, where individuals within groups tend to copy a "leader" (or teacher), altruistic helping can evolve in groups of any size, although the cultural transmission rule itself hitchhikes rather weakly with a selected helping trait. Our results suggest that culturally determined helping behaviors are more likely to be driven by "leaders" than by popularity, but the emergence and stability of the cultural transmission rules themselves should be driven by some extrinsic factors.

A New Approach for the Carbon Monoxide (CO) Exposure Diagnosis: Measurement of Total CO in Human Blood Versus Carboxyhemoglobin (HbCO).

The aim of the study is to present the application of a headspace-gas chromatography-mass spectrometry (HS-GC-MS) method for the determination of the carbon monoxide (CO) blood concentration and to compare it with carboxyhemoglobin (HbCO) saturation. In postmortem cases, the HbCO measured by spectrophotometry frequently leads to inaccurate results due to inadequate samples or analyses. The true role of CO intoxication in the death of a person could be misclassified. The estimation of HbCO from HS-GC-MS CO measurements provides helpful information by determining the total CO levels (CO linked to hemoglobin (HbCO) and CO dissociated from hemoglobin). The CO concentrations were converted in HbCO saturation levels to define cutoff blood CO values. CO limits were defined as less than 1 μmol/mL for living persons, less than 1.5 μmol/mL for dead persons without CO exposure, and greater than 3 μmol/mL for dead persons with clear CO poisoning.

L'art. 320 al. 2 CO: une règle compatible avec les dispositions sur la conclusion des contrats?

Le Tribunal fédéral et la doctrine majoritaire interprètent l'art. 320 al. 2 CO comme un cas où l'existence d'un contrat de travail peut être déduite de circonstances de fait indépendantes de tout consensus. Il s'agirait donc là d'une dérogation aux art. 1 ss CO. Dans cette contribution, nous proposons une conception de l'art. 320 al. 2 CO compatible avec le mécanisme ordinaire de conclusion des contrats. En l'occurrence, le contrat de travail serait conclu par actes concluants sur la base d'un accord de droit, c'est-à-dire en appliquant le principe de la confiance. Alors que la lettre et le but de protection de l'art. 320 al. 2 CO semblent plutôt neutres à cet égard, l'histoire et la systématique de cet alinéa plaident clairement en faveur de la conception ici proposée.

The effect of n-3 enriched nutrition therapy on post-operative cognitive dysfunction after cardiac surgery

Background: Postoperative cognitive dysfunction (POCD) occurs frequently after cardiac surgery. Some data suggest that inflammation plays a key role in the development of POCD. N-3 fatty acids have been shown to have a beneficial effect on inflammation. We hypothesised that perioperative n-3 enriched nutrition therapy would reduce the incidence of POCD in this group of patients. Methods: Randomized, double blind placebo controlled trial in patients aged 65 or older undergoing elective cardiac surgery with cardiopulmonary bypass. 2x 250 mL placebo (Ensure Plus™, Abbott Nutrition) or n-3 enriched nutrition therapy (ProSure™ Abbott Nutrition) were administered for ten days starting 5 days prior to surgery. Cognition was assessed preoperatively and 7 days after surgery with the Consortium to Establish a Registry for Alzheimer's Disease - Neuropsychological Assessment Battery (CERAD-NAB) [1]. Results: 16 patients were included. Mean age was 72 } 5.3 for placebo and 75 } 4.8 for ProSure™ respectively. CRP and IL-6 did not differ significantly between groups preoperatively and on postoperative days 1, 3, and 7. Preoperative CERAD total scores were 86 } 10 and 81 } 9 (p = n.s.) for Placebo and ProSure™, respectively. Postoperative scores were 88 } 12, and 77 } 19 (p = n.s.) The change in score was not different between the two groups (Placebo: +3 } 5; ProSure: -5 } 11). Conclusion: In this very small sample no effect of preoperatively started n-3 enriched nutritional supplements on inflammation or cognitive functions were detected. However, there is a large likelihood of a type II error and more patients need to be included to assess possible beneficial effects of this intervention in elderly patients undergoing elective cardiac surgery. 1 Chandler MJ, et al. Neurology. 2005;65:102-6.

De Novo Production of Metabolites by Fungal Co-culture of Trichophyton rubrum and Bionectria ochroleuca.

The co-cultivation of fungi has recently been described as a promising strategy to induce the production of novel metabolites through possible gene activation. A large screening of fungal co-cultures in solid media has identified an unusual long-distance growth inhibition between Trichophyton rubrum and Bionectria ochroleuca. To study metabolite induction in this particular fungal interaction, differential LC-MS-based metabolomics was performed on pure strain cultures and on their co-cultures. The comparison of the resulting fingerprints highlighted five de novo induced compounds, which were purified using software-oriented semipreparative HPLC-MS. One metabolite was successfully identified as 4″-hydroxysulfoxy-2,2″-dimethylthielavin P (a substituted trimer of 3,5-dimethylorsellinic acid). The nonsulfated form, as well as three other related compounds, were found in the pure strain culture of B. ochroleuca.

Avian haemosporidian persistence and co-infection in great tits at the individual level.

BACKGROUND: Many studies have tracked the distribution and persistence of avian haemosporidian communities across space and time at the population level, but few studies have investigated these aspects of infection at the individual level over time. Important aspects of parasite infection at the individual level can be missed if only trends at the population level are studied. This study aimed to determine how persistent Haemosporida are in great tit individuals recaptured over several years, whether parasitaemia differed by parasite lineage (mitochondrial cytochrome b haplotype) and how co-infection (i.e. concurrent infection with multiple genera of parasites) affects parasitaemia and body mass. METHODS: Parasite prevalence was determined by polymerase chain reaction (PCR), quantitative PCR were used to assess parasitaemia and sequencing was employed to determine the identity of the lineages using the MalAvi database. RESULTS: Haemosporidian prevalence was high over sampled years with 98% of 55 recaptured individuals showing infection in at least one year of capture. Eighty-two percent of all positive individuals suffered co-infection, with an overall haemosporidian lineage diversity of seventeen. Plasmodium and Haemoproteus parasites were found to be highly persistent, with lineages from these genera consistently found in individuals across years and with no differences in individual parasitaemia being recorded at subsequent captures. Conversely, Leucocytozoon parasites showed higher turnover with regard to lineage changes or transitions in infection status (infected vs non-infected) across years. Parasitaemia was found to be lineage specific and there was no relationship between Plasmodium parasitaemia or host body condition and the presence of Leucocytozoon parasites. CONCLUSIONS: The findings of this study suggest that different genera of haemosporidian parasites interact differently with their host and other co-infecting parasites, influencing parasite persistence most likely through inter-parasite competition or host-parasite immune interactions. Even-though co-infections do not seem to result in increased virulence (higher parasitaemia or poorer host body condition), further investigation into infection potential of these parasites, both individually and as co-infections, is necessary.

Detection of metabolite induction in fungal co-cultures on solid media by high-throughput differential ultra-high pressure liquid chromatography-time-of-flight mass spectrometry fingerprinting.

Access to new biological sources is a key element of natural product research. A particularly large number of biologically active molecules have been found to originate from microorganisms. Very recently, the use of fungal co-culture to activate the silent genes involved in metabolite biosynthesis was found to be a successful method for the induction of new compounds. However, the detection and identification of the induced metabolites in the confrontation zone where fungi interact remain very challenging. To tackle this issue, a high-throughput UHPLC-TOF-MS-based metabolomic approach has been developed for the screening of fungal co-cultures in solid media at the petri dish level. The metabolites that were overexpressed because of fungal interactions were highlighted by comparing the LC-MS data obtained from the co-cultures and their corresponding mono-cultures. This comparison was achieved by subjecting automatically generated peak lists to statistical treatments. This strategy has been applied to more than 600 co-culture experiments that mainly involved fungal strains from the Fusarium genera, although experiments were also completed with a selection of several other filamentous fungi. This strategy was found to provide satisfactory repeatability and was used to detect the biomarkers of fungal induction in a large panel of filamentous fungi. This study demonstrates that co-culture results in consistent induction of potentially new metabolites.