Whole-cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Many high-value added recombinant proteins, such as therapeutic glycoproteins, are produced using mammalian cell cultures. In order to optimise the productivity of these cultures it is important to monitor cellular metabolism, for example the utilisation of nutrients and the accumulation of metabolic waste products. One metabolic waste product of interest is lactic acid (lactate), overaccumulation of which can decrease cellular growth and protein production. Current methods for the detection of lactate are limited in terms of cost, sensitivity, and robustness. Therefore, we developed a whole-cell Escherichia coli lactate biosensor based on the lldPRD operon and successfully used it to monitor lactate concentration in mammalian cell cultures. Using real samples and analytical validation we demonstrate that our biosensor can be used for absolute quantification of metabolites in complex samples with high accuracy, sensitivity and robustness. Importantly, our whole-cell biosensor was able to detect lactate at concentrations more than two orders of magnitude lower than the industry standard method, making it useful for monitoring lactate concentrations in early phase culture. Given the importance of lactate in a variety of both industrial and clinical contexts we anticipate that our whole-cell biosensor can be used to address a range of interesting biological questions. It also serves as a blueprint for how to capitalise on the wealth of genetic operons for metabolite sensing available in Nature for the development of other whole-cell biosensors

An Integrated Accident & Consequence Analysis Approach for Accidental Releases through Multiple Leak Paths

This paper presents a consequence analysis for a postulated fire accident on a building containing plutonium when the resulting outside release is partly through the ventilation/filtration system and partly through other pathways such as building access doorways. When analyzing an accident scenario involving the release of radioactive powders inside a building, various pathways for the release to the outside environment can exist. This study is presented to guide the analyst on how the multiple building leak path factors (combination of filtered and unfiltered releases) can be evaluated in an integrated manner starting with the source term calculation and proceeding through the receptor consequence determination. The analysis is performed in a two-step process. The first step of the analysis is to calculate the leak path factor, which represents the fraction of respirable radioactive powder that is made airborne that leaves the building through the various pathways. The computer cod e of choice for this determination is MELCOR. The second step is to model the transport and dispersion of powder material released to the atmosphere and to estimate the resulting dose that is received by the downwind receptors of interest. The MACCS computer code is chosen for this part of the analysis. This work can be used as model for performing analyses for systems similar in nature where releases can propagate to the outside environment via filtered and unfiltered pathways. The methodology provides guidance to analysts outlining the essential steps needed to perform a sound and defensible consequence analysis

A Density Matrix-based Algorithm for Solving Eigenvalue Problems

A new numerical algorithm for solving the symmetric eigenvalue problem is presented. The technique deviates fundamentally from the traditional Krylov subspace iteration based techniques (Arnoldi and Lanczos algorithms) or other Davidson-Jacobi techniques, and takes its inspiration from the contour integration and density matrix representation in quantum mechanics. It will be shown that this new algorithm - named FEAST - exhibits high efficiency, robustness, accuracy and scalability on parallel architectures. Examples from electronic structure calculations of Carbon nanotubes (CNT) are presented, and numerical performances and capabilities are discussed.Comment: 7 pages, 3 figure

ORAL LESIONS DUE TO ACINETOBACTER BAUMANNII INFECTION IN A PATIENT AFFECTED BY AUTOIMMUNE HEMOLYTIC ANEMIA (AIHA): FIRST CLINICAL REPORT

Aim. Acinetobacter Baumannii (coming from the Greek “akinetos,” i.e. non-motile) is an opportunistic bacterial pathogen primarily associated with hospital-acquired infections. Commonly associated with aquatic environments, A. Baumannii easily contaminates the surrounding environment and it colonizes acute ill patients in which can survive for several days. Generally, A. Baumannii is able to damage mucous membranes or exposed skin after accident or injury; it may be responsible of many diseases such as pneumonia, bacteremia, meningitis, urinary tract infections, peritonitis and infections of skin and soft tissues. Tissues infected by A. Baumannii initially present “orange peel” appearance followed by sandpaper-like presentation, when there is a disruption, hemorrhagic bullae can be seen with a visible necrotizing process followed by bacteremia. Current therapy is based on intravenous administration of tigecycline 100-200 mg (first dose) and 50-100 mg every 12 h for up to 14 days, unless complications. If untreated, this infection can lead to septicemia and death. The mortality rate of this infection is high, especially in case of bacteremia (52%) and pneumonia (23–73%). A. Baumannii is resistant to many drugs and represents an important nosocomial pathogen that particularly infects critically ill patients. At the best of our knowledge, no case of oral infection has been reported. to present the first one case characterized by oral soft tissue infection due to A. Baumannii responsive to imipenem. Materials and methods. a 78 years male was hospitalized in August of 2013 at the Hematology unit of the A.O.U.P. “P. Giaccone” of Palermo with a diagnosis of autoimmune hemolytic anemia (AIHA). He was treated per os with corticosteroids (Prednisone) and immunosuppressants (Rituximab and Cyclophosphamide), also IVIG (Intra Venous Immuno-Globulin) was administreted. In November 2013, the patient was treated with piperacillin IV (Tazocin) for treating an urinary tract infection; four days after, multiple oral ulcerative bullous lesions on the lingual and buccal mucosa and crusted lesions on the lip vermilion appeared, associated with intense pain. An oral swab for bacteria research was carried out; topical therapy (chlorhexidine rinses and hyaluronic acid gel) has been prescribed and piperacillin therapy was stopped. Oral swab outcome resulted positive for A. Baumannii and Enterococcus Faecalis, both sensitive only to imipenem, that was administrated (500 mg IV every 8h) for 10 days. The patient was immediately isolated in a single room for preventing and controlling the spread of A. Baumannii. Results. From diagnosis, every 3 days clinical examination of the oral cavity was performed, revealing the progressive regression within thirty days until complete healing without leaving scars. After, a second oral swab confirmed the absence of any bacteria. Conclusions. The World Health Organization has recently identified antimicrobial resistance as one of the three most important problems facing human health and among the most common and serious pathogens, including A. Baumannii. It is an emerging potentially drug-resistant micro-organism and its isolation must alert physicians to carry on all preventive measures for avoiding contamination of other patients, especially those immunosuppressed, at risk for severe persistent infections or death. This precaution should be continued for all the duration of hospitalization and until the negativization of culture samples was obtained. It is important that physicians and dentists recognize suspicious lesions in unusual locations, such as oral mucosa, in absence of other known etiological factors in a timely manner before the diffusion among other patients in order to avoid the spread of a nosocomial outbreak

Calonectria spp. causing leaf spot, crown and root rot of ornamental plants in Tunisia

Calonectria spp. are important pathogens of ornamental plants in nurseries, especially in the Northern Hemisphere. They are commonly associated with a wide range of disease symptoms of roots, leaves and shoots. During a recent survey in Tunisia, a number of Calonectria spp. were isolated from tissues of ornamental plants showing symptoms of leaf spot, crown and root rot. The aim of this study was to identify these Calonectria spp. using morphological and DNA sequence comparisons. Two previously undescribed Calonectria spp., C. pseudomexicana sp. nov. and C. tunisiana sp. nov., were recognised. Calonectria mexicana and C. polizzii are newly reported for the African continent. Pathogenicity tests with all four Calonectria spp. showed that they are able to cause disease on seedlings of Callistemon spp., Dodonaea viscosa, Metrosideros spp. and Myrtus communis

The influence of social support on the wellbeing of immigrants residing in Italy: Sources and functions as predictive factors for life satisfaction levels, sense of community and resilience

Moving from one country to another involves not only separation from the country of origin, but also the tiring process of integration into a new physical, institutional, and sociocultural context, which may expose migrants to acculturation stress. The loss of former support networks, or at the very least their transformation, presents immigrants with the need to rebuild their social support systems in the host country, involving an active search for support. Therefore, the aim of study is to analyze the structure of informal social support and its capacity to predict immigrants’ sense of community, resilience, and satisfaction with life. The results confirm that social support predicts satisfaction with life, sense of community, and resilience. Our findings highlight the way sources and frequency of support, and the satisfaction with which they are associated, have different degrees of predictive value on the dependent variables under investigation. In this study, it can be concluded that social support is an important factor in the well-being of migrants and their integration into the host community. The results have an important practical value in promoting interventions that improve immigrants’ support networks and, consequently, increase their satisfaction with life, sense of community, and resilience

The influence of social support on the wellbeing of immigrants residing in Italy: Sources and functions as predictive factors for life satisfaction levels, sense of community and resilience

Moving from one country to another involves not only separation from the country of origin, but also the tiring process of integration into a new physical, institutional, and sociocultural context, which may expose migrants to acculturation stress. The loss of former support networks, or at the very least their transformation, presents immigrants with the need to rebuild their social support systems in the host country, involving an active search for support. Therefore, the aim of study is to analyze the structure of informal social support and its capacity to predict immigrants’ sense of community, resilience, and satisfaction with life. The results confirm that social support predicts satisfaction with life, sense of community, and resilience. Our findings highlight the way sources and frequency of support, and the satisfaction with which they are associated, have different degrees of predictive value on the dependent variables under investigation. In this study, it can be concluded that social support is an important factor in the well-being of migrants and their integration into the host community. The results have an important practical value in promoting interventions that improve immigrants’ support networks and, consequently, increase their satisfaction with life, sense of community, and resilience.Funding for open access charge: Universidad de Málaga / CBU

Evidence that muscle cells do not express the histidine-rich glycoprotein associated with AMP deaminase but can internalise the plasma protein

Histidine-rich glycoprotein (HRG) is synthesized by liver and is present at relatively high concentration in the plasma of vertebrates. We have previously described the association of a HRG-like molecule to purified rabbit skeletal muscle AMP deaminase (AMPD). We also provided the first evidence for the presence of a HRG-like protein in human skeletal muscle where a positive correlation between HRG content and total determined AMPD activity has been shown. In the present paper we investigate the origin of skeletal muscle HRG. The screening of a human skeletal muscle cDNA expression library using an anti-HRG antibody failed to reveal any positive clone. The RT-PCR analysis, performed on human skeletal muscle RNA as well as on RNA from the rhabdomyosarcoma (RD) cell line, failed to show any mRNA specific for the plasma HRG or for the putative muscle variant. When the RD cells were incubated with human plasma HRG, a time-dependent increase of the HRG immunoreactivity was detected both at the plasma membrane level and intracellularly. The internalisation of HRG was inhibited by the addition of heparin. The above data strongly suggest that skeletal muscle cells do not synthesize the muscle variant of HRG but instead can actively internalise it from plasma

Method and apparatus for electromagnetic powder deposition

The present invention provides a method for depositing powder particles on a substrate. The method comprises forming a planar plasma armature, accelerating the plasma armature, accelerating a column of gas with the plasma armature; and accelerating the powder particles with the column of gas. The present invention provides for a railgun, comprising first and second conducting rails, and first and second insulating rails. The insulating and conducting rails form a bore of the railgun. The first and second conducting rails are separated by the insulating rails. At least one of the rails has a port in the wall thereof, the port is adapted to introducing powder particles into the bore.Board of Regents, University of Texas Syste