Studies on processing, particle formation, and immunogenicity of the HIV-1 gag gene product: a possible component of a HIV vaccine

Antigens in a particulate conformation were shown to be highly immunogenic in mammals. For this reason, the particle forming capacity of derivatives of the HIV-1 group specific core antigen p55 gag was assayed and compared dependent on various expression systems: recombinant bacteria, vaccinia- and baculoviruses were established encoding the entire core protein p55 either in its authentic sequence or lacking the myristylation consensus signal. Moreover, p55 gag was expressed in combination with the protease (p55-PR) or with the entire polymerase (p55-pol), respectively. Budding of 100-160 nm p55 core particles, resembling immature HIV-virions, was observed in the eucaryotic expression systems only. In comparison to the vaccinia virus driven expression of p55 in mammalian cells, considerably higher yields of particulate core antigen were obtained by infection of Spodoptera frugiperda (Sf9) insect cells with the recombinant Autographa californica nuclear polyhedrosis (AcMNPV) baculovirus. Mutation of the NH2-terminal myristylation signal sequence prevented budding of the immature core particles. Expression of the HIV p55-PR gene construct by recombinant baculovirus resulted in complete processing of the p55 gag precursor molecule in this system. The introduction of an artificial frameshift near the natural frameshift site resulted in constitutive expression of the viral protease and complete processing of p55, both in Escherichia coli and in vaccinia virus infected cells. Interestingly, significant processing of p55 resembling that of HIV infected H9 cells could also be achieved in the vaccinia system by fusing the entire pol gene to the gag gene. Moreover, processing was not found to be dependent on amino-terminal myristylation of the gag procursor molecule, which is in contrast to observations with type C and type D retrovirus. However, complete processing of p55 into p24, p17, p9 and p6 abolished particle formation. Purified immature HIV-virus like particles were highly immunogenic in rabbits, leading to a strong humoral immune response after immunization. Empty immature p55 gag particles represent a noninfectious and attractive candidate for a basic vaccine component

Extensions of adaptive slope-seeking for active flow control

Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG geförderten) Allianz- bzw. Nationallizenz frei zugänglich.This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.To speed up gradient estimation in a slope-seeking controller two different modifications are proposed in this study. In a first approach, the gradient estimation is based on a locally identified black-box model. A further improvement is obtained by applying an extended Kalman filter to estimate the local gradient of an input—output map. Moreover, a simple method is outlined to adapt the search radius in the classical extremum- and slope-seeking approach to reduce the perturbations near the optimal state. To show the versatility of the slope-seeking controller for flow control applications two different wind tunnel experiments are considered, namely with a two-dimensional bluff body and a generic three-dimensional car model (Ahmed body).DFG, SFB 557, Beeinflussung komplexer turbulenter Scherströmunge

Kinetic models for dilute solutions of dumbbells in non-homogeneous flows revisited

We propose a two fluid theory to model a dilute polymer solution assuming that it consists of two phases, polymer and solvent, with two distinct macroscopic velocities. The solvent phase velocity is governed by the macroscopic Navier-Stokes equations with the addition of a force term describing the interaction between the two phases. The polymer phase is described on the mesoscopic level using a dumbbell model and its macroscopic velocity is obtained through averaging. We start by writing down the full phase-space distribution function for the dumbbells and then obtain the inertialess limits for the Fokker-Planck equation and for the averaged friction force acting between the phases from a rigorous asymptotic analysis. The resulting equations are relevant to the modelling of strongly non-homogeneous flows, while the standard kinetic model is recovered in the locally homogeneous case

The SARS-coronavirus-host interactome

Coronaviruses (CoVs) are important human and animal pathogens that induce fatal respiratory, gastrointestinal and neurological disease. The outbreak of the severe acute respiratory syndrome (SARS) in 2002/2003 has demonstrated human vulnerability to (Coronavirus) CoV epidemics. Neither vaccines nor therapeutics are available against human and animal CoVs. Knowledge of host cell proteins that take part in pivotal virus-host interactions could define broad-spectrum antiviral targets. In this study, we used a systems biology approach employing a genome-wide yeast-two hybrid interaction screen to identify immunopilins (PPIA, PPIB, PPIH, PPIG, FKBP1A, FKBP1B) as interaction partners of the CoV non-structural protein 1 (Nsp1). These molecules modulate the Calcineurin/NFAT pathway that plays an important role in immune cell activation. Overexpression of NSP1 and infection with live SARS-CoV strongly increased signalling through the Calcineurin/NFAT pathway and enhanced the induction of interleukin 2, compatible with late-stage immunopathogenicity and long-term cytokine dysregulation as observed in severe SARS cases. Conversely, inhibition of cyclophilins by cyclosporine A (CspA) blocked the replication of CoVs of all genera, including SARS-CoV, human CoV-229E and -NL-63, feline CoV, as well as avian infectious bronchitis virus. Non-immunosuppressive derivatives of CspA might serve as broad-range CoV inhibitors applicable against emerging CoVs as well as ubiquitous pathogens of humans and livestock

Volumetrie der Schilddrüsenlappen mittels Real-time-Sonographie

Im direkten Vergleich mit der durch Submersion ermittelten wahren Schilddrüsengröße wurde bei Leichen mit einem Real-time-Gerät ein Verfahren zur Volumenbestimmung durch sonographische Messungen erarbeitet. Länge mal Breite mal Dicke des Schilddrüsenlappens, multipliziert mit dem Faktor π/6, entsprechen einem Rotationsellipsoid, während sich bei Multiplikation mit dem optimierten Korrekturfaktor f = 0,479 das bestmögliche rechnerische Volumen des Lappens ergibt. Die Richtigkeit dieser Bestimmung ist definitionsgemäß 100 %; der. durchschnittliche Fehler der Methode beträgt 16 %. Die Messungen sind einfach durchzuführen und verlangen keinerlei Zusatzeinrichtungen für Planimetrie oder Rechneroperationen. Die Volumetrie der Schilddrüse ist vor allem zur Therapiebeurteilung und zur Berechnung der Dosis bei einer Radiojodtherapie erforderlich.Thyroid volume as measured by real-time ultrasound in cadavers was compared with direct measurements obtained by submersion. Length × width × thickness of the thyroid lobe multiplied by factor π/6, correspond to a rotation ellipsoid, while the best calculated volume of the lobe is obtained by multiplying with the optimised correction factor f = 0.479. The correctness of this calculation is, by definition, 100 %; average error of the method is 16 %. The measurements are easy to do and require no additional equipment for planimetry or calculations. Volumetric analysis of the thyroid gland is especially necessary in assessing results of treatment and for measuring dosage in connection with radioiodine therapy

High (but Not Low) Urinary Iodine Excretion Is Predicted by Iodine Excretion Levels from Five Years Ago

Background: It has not been investigated whether there are associations between urinary iodine (UI) excretion measurements some years apart, nor whether such an association remains after adjustment for nutritional habits. The aim of the present study was to investigate the relation between iodine-creatinine ratio (ICR) at two measuring points 5 years apart. Methods: Data from 2,659 individuals from the Study of Health in Pomerania were analyzed. Analysis of covariance and Poisson regressions were used to associate baseline with follow-up ICR. Results: Baseline ICR was associated with follow-up ICR. Particularly, baseline ICR >300 mu g/g was related to an ICR >300 mu g/g at follow-up (relative risk, RR: 2.20; p < 0.001). The association was stronger in males (RR: 2.64; p < 0.001) than in females (RR: 1.64; p = 0.007). In contrast, baseline ICR <100 mu g/g was only associated with an ICR <100 mu g/g at follow-up in males when considering unadjusted ICR. Conclusions: We detected only a weak correlation with respect to low ICR. Studies assessing iodine status in a population should take into account that an individual with a low UI excretion in one measurement is not necessarily permanently iodine deficient. On the other hand, current high ICR could have been predicted by high ICR 5 years ago. Copyright (C) 2011 S. Karger AG, Base

The balance of mental health care in Europe: a comparative analysis of core health care versus the provision of other types of care for adults with mental health problems in eight study areas

Aims Although many mental health care systems provide care interventions that are not related to direct health care, little is known about the interfaces between the latter and core health care. ‘Core health care’ refers to services whose explicit aim is direct clinical treatment which is usually provided by health professionals, i.e., physicians, nurses, psychologists. ‘Other care’ is typically provided by other staff and includes accommodation, training, promotion of independence, employment support and social skills. In such a definition, ‘other care’ does not necessarily mean being funded or governed differently. The aims of the study were: (1) using a standard classification system (Description and Evaluation of Services and Directories in Europe for Long Term Care, DESDE-LTC) to identify ‘core health’ and ‘other care’ services provided to adults with mental health problems; and (2) to investigate the balance of care by analysing the types and characteristics of core health and other care services. Methods The study was conducted in eight selected local areas in eight European countries with different mental health systems. All publicly funded mental health services, regardless of the funding agency, for people over 18 years old were identified and coded. The availability, capacity and the workforce of the local mental health services were described using their functional main activity or ‘Main Types of Care’ (MTC) as the standard for international comparison, following the DESDE-LTC system. Results In these European study areas, 822 MTCs were identified as providing core health care and 448 provided other types of care. Even though one-third of mental health services in the selected study areas provided interventions that were coded as ‘other care’, significant variation was found in the typology and characteristics of these services across the eight study areas. Conclusions The functional distinction between core health and other care overcomes the traditional division between ‘health’ and ‘social’ sectors based on governance and funding. The overall balance between core health and other care services varied significantly across the European sites. Mental health systems cannot be understood or planned without taking into account the availability and capacity of all services specifically available for this target population, including those outside the health sector

Identification of a protein encoded in the EB-viral open reading frame BMRF2

Using monospecific rabbit sera against a peptide derived from a potential antigenic region of the Epstein-Barr viral amino acid sequence encoded in the open reading frame BMRF2 we could identify a protein-complex of 53/55 kDa in chemically induced B95-8, P3HR1 and Raji cell lines. This protein could be shown to be membrane-associated, as predicted by previous computer analysis of the secondary structure and hydrophilicity pattern, and may be a member of EBV-induced membrane proteins in lytically infected cells

Anti-CD117 CAR T cells incorporating a safety switch eradicate human acute myeloid leukemia and hematopoietic stem cells

Discrimination between hematopoietic stem cells and leukemic stem cells remains a major challenge for acute myeloid leukemia immunotherapy. CAR T cells specific for the CD117 antigen can deplete malignant and healthy hematopoietic stem cells before consolidation with allogeneic hematopoietic stem cell transplantation in absence of cytotoxic conditioning. Here we exploit non-viral technology to achieve early termination of CAR T cell activity to prevent incoming graft rejection. Transient expression of an anti-CD117 CAR by mRNA conferred T cells the ability to eliminate CD117+ targets in vitro and in vivo. As an alternative approach, we used a Sleeping Beauty transposon vector for the generation of CAR T cells incorporating an inducible Caspase 9 safety switch. Stable CAR expression was associated with high proportion of T memory stem cells, low levels of exhaustion markers, and potent cellular cytotoxicity. Anti-CD117 CAR T cells mediated depletion of leukemic cells and healthy hematopoietic stem cells in NSG mice reconstituted with human leukemia or CD34+ cord blood cells, respectively, and could be terminated in vivo. The use of a non-viral technology to control CAR T cell pharmacokinetic properties is attractive for a first-in-human study in patients with acute myeloid leukemia prior to hematopoietic stem cell transplantation