RESUMO
Residents and staff of nursing homes are important target groups for influenza vaccination in Germany. The aim of this study was to gain the first insights into whether nursing homes organize activities with respect to vaccination against influenza and whether there is a demand for further information. In the context of the national influenza immunization campaign-which is jointly carried out by the Robert Koch Institute (RKI) and the Federal Centre for Health Education (BZgA) on an annual basis-influenza information kits were sent to the management of 10,700 nursing homes in September 2013. Along with the information material, the institutions also received a questionnaire to which they were able to respond via mail, fax, or online. Data from 988 homes were included in the analysis. The majority of institutions informed both residents (88.9 %) and nursing staff (81.2 %) about influenza vaccination. However, only 64.7 % of nursing homes carried out specific immunization activities for their residents and only half (49.3 %) offered a flu shot to their staff. When asked why the institutions do not provide influenza-specific information and vaccination to their staff, the majority had the opinion that this is the responsibility of each individual's general practitioner. Overall, only 4.9 % of nursing homes assessed influenza vaccination coverage among their staff annually. A third of all surveyed institutions (33.6 %) expressed a demand for additional influenza vaccine-related information. In conclusion, improved health education is needed to raise awareness about the importance of influenza vaccination among residents and employees of nursing homes in Germany so as to prevent influenza-associated morbidity and mortality in this risk group.
Assuntos
Promoção da Saúde/estatística & dados numéricos , Vacinas contra Influenza/uso terapêutico , Influenza Humana/prevenção & controle , Avaliação das Necessidades , Enfermeiras e Enfermeiros/estatística & dados numéricos , Casas de Saúde/estatística & dados numéricos , Vacinação/estatística & dados numéricos , Adulto , Idoso , Feminino , Alemanha/epidemiologia , Pesquisas sobre Atenção à Saúde , Humanos , Influenza Humana/epidemiologia , Masculino , Pessoa de Meia-Idade , Padrões de Prática em Enfermagem/estatística & dados numéricosRESUMO
A feasibility study using an ion mobility spectrometer coupled with a multi-capillary column (MCC) was started to identify characteristic peaks of volatile compounds in exhaled human breath samples of 10 ml volume. The breath of 20 patients with sarcoidosis and suspicion of sarcoidosis because of mediastinal lymph node enlargement was investigated. Using a set of procedures for data processing and scoring a sector of interest was determined within the IMS-chromatogram. It could be shown that a procedure related to a single peak in the IMS-chromatogram delivers differentiation into the two groups of patients with confirmed sarcoidosis and those suffering no sarcoidosis. The potential biomarker is characterized by the following parameters: inverse mobility (1/K(0)) 0.53 ± 0.01 V s cm(-2)-retention time 22 ± 5 s. These results are a first step in breath analysis by MCC/IMS in patients with sarcoidosis by an automated procedure applied to IMS-chromatograms directly.
RESUMO
For the production of a chemically inactivated Parapoxvirus ovis (PPVO), an adherent bovine kidney cell line was cultivated on Cytodex-3 microcarriers in suspension culture. The inactivated and purified virus particles have shown immune modulatory activity in several animal models. PPVO was produced by a biphasic batch process at the 3.5 and 10 L scale. Aeration was realised by bubble-free membrane oxygenation via a tube stator with a central two-blade anchor impeller. In order to increase efficiency, process robustness and safety, the established process was optimised. The cell line was adapted to a protein-free medium (except recombinant insulin) in order to increase biosafety. A scale up to a 50 L pilot plant with direct cell expansion was performed successfully. In parallel, the biphasic batch process was optimised with special emphasis on different operating conditions (cell number, Multiplicity of Infection (MOI), etc.) and process management (fed-batch, dialysis, etc.). The quality and concentration of the purified virus particles was assessed by quantitative electron microscopy, residual host cell protein and DNA-content and, finally, biologic activity in a transgenic mouse model. This integrated approach led to a new, safe, robust and highly productive large-scale production process, called "Volume-Expanded-Fed" Batch with cell densities up to 6-7e06 cells/mL. By subsequent dilution of infected cells into the next process scale, an increase in total productivity by a factor of 40 (related to an established biphasic batch process) was achieved.
Assuntos
Parapoxvirus/crescimento & desenvolvimento , Parapoxvirus/imunologia , Vacinas de Produtos Inativados/biossíntese , Animais , Bovinos , Contagem de Células , Sobrevivência Celular , Células Cultivadas , Meios de Cultura , DNA Viral/biossíntese , DNA Viral/imunologia , Hepatite B/imunologia , Rim/citologia , Rim/metabolismo , Camundongos , Camundongos Transgênicos , Microscopia Eletrônica , Tripsina/química , Vacinas de Produtos Inativados/imunologiaRESUMO
An EBV-transformed human B cell line, which produces monoclonal IgM antibodies, was cultured in an immobilized state on a ceramic matrix in the Opticell system. Cell growth, metabolic activity and product formation of these cells were optimized in a single semi-continuous culture of 10 inductions for 29 d. All in all, 19.5 g of IgM were harvested in 193 l of culture supernatant. The average production of IgM in the Opticell unit of 20 l was 0.7 g/d which was obtained even in serum-reduced medium (1%). Under optimal conditions IgM yields were enhanced to 1.5-2.0 g/d. These results indicate that the Opticell is a suitable system for the large scale production of monoclonal antibodies, particularly because the capacity for aeration and pumping of one Opticell unit is sufficient to control 3 growth chambers running in parallel.
Assuntos
Anticorpos Monoclonais/isolamento & purificação , Biotecnologia/métodos , Adesão Celular , Células Cultivadas , Cerâmica , Glucose/metabolismo , Glutamina/metabolismo , Humanos , Hibridomas/fisiologia , Imunoglobulina M/isolamento & purificação , Consumo de OxigênioRESUMO
The influence of a protein-free and a protein-rich, supplemented serum-free medium on the production of plasminogen activator (t-PA) from Bowes melanoma cells was investigated in the Opticell culture system and compared to tissue culture flask cultures. In the presence of medium supplements metabolic activity and t-PA production were favoured in both systems. The addition of supplements was apparently more effective in the Opticell than in flask cultures, because t-PA activity obtained in the Opticell was 2-3 times higher in protein-rich medium, but 2 times lower in unsupplemented medium than in flasks. These results indicate that the protein content in a serum-free medium is important for product formation in the Opticell, and serum-free media which work at small scale in tissue culture flasks are not always suited for technical culture systems such as the Opticell but have to be adapted to them.
Assuntos
Melanoma Experimental/patologia , Ativador de Plasminogênio Tecidual/biossíntese , Biotecnologia/instrumentação , Sangue , Adesão Celular , Linhagem Celular , Células Cultivadas , Meios de Cultura , Glucose/metabolismo , Glutamina/metabolismo , Humanos , Serina/metabolismoRESUMO
A screening method was established to develop chemically defined, serum-free culture media for human adherent cell lines. It indicates within 2-6 months whether a certain cell line is suited for routine cultivation under serum-free conditions, and which medium supplements and surface coatings of the culture vessels the cells need for minimal demand. This method was tested with the human cell lines Bowes melanoma and prostate carcinoma 3 (PC 3).
Assuntos
Adesão Celular , Linhagem Celular , Meios de Cultura , Divisão Celular , Estudos de Avaliação como Assunto , Humanos , Cinética , Melanoma/patologiaRESUMO
Cultivation parameters for the production of five lymphokines, granulocyte-macrophage colony stimulating factor (GM-CSF), interferon-gamma (IFN-gamma), interleukin 2 (IL-2), macrophage cytotoxicity factor (MCF), and macrophage migration inhibitory factor (MIF) from human spleen cells or peripheral blood lymphocytes were optimized. Cultivation was done in bioreactors containing up to 200 ml of medium, usually serum-free. The reactors were equipped with surface aeration facilities, stirrers and oxygen electrodes. Whereas stirring speed alone did not influence the yields of lymphokines, good aeration was especially beneficial for high IL-2 yields. However, all lymphokines were also produced under anaerobic conditions. The concentration of the mitogen concanavalin A was mainly critical for optimal IL-2 release. Optimal cell concentrations varied from 5 X 10(6)/ml (for GM-GSF and MCF) to 10 X 10(6)/ml (for IL-2 and IFN-gamma). It was possible to increase the yields of individual lymphokines 3 to 10-fold per batch of lymphocytes by a reinduction procedure which involved a change of medium and mitogen every 24 hrs. Reinduction was possible up to 4 times, especially when serum was present in the culture media.
Assuntos
Concanavalina A/farmacologia , Ativação Linfocitária , Linfocinas/biossíntese , Células Cultivadas , Fatores Estimuladores de Colônias/biossíntese , Humanos , Interferon gama/biossíntese , Interleucina-2/biossíntese , Linfócitos/imunologia , Fatores Inibidores da Migração de Macrófagos/biossíntese , Macrófagos/imunologia , Oxigênio/farmacologia , Baço/citologiaRESUMO
The production of the lymphokines colony stimulating factor (CSF), interleukin-2 (IL-2), macrophage cytotoxicity factor (MCF), macrophage migration inhibitory factor (MIF) and T cell replacing factor (TRF) was optimized from mouse spleen cell cultures stimulated with concanavalin A (Con A). The cultivation was performed in bioreactors which allow regulation of dissolved oxygen concentration. The oxygen supply influenced the yields of individual lymphokines in different ways. High oxygen concentration was beneficial for high release of IL-2 and MCF, whereas MIF and TRF were better produced at low oxygen concentrations. CSF release did not depend on oxygen supply. Upon stimulation with Con A, lymphokine activities in the culture supernatant reached an optimum or plateau after 24 h Lymphocytes continued with lymphokine release, when they were restimulated for further 24 h. in fresh medium and mitogen. CSF could be induced for at least 4 times. MCF was released during the first 3 inductions, whereas IL-2 and TRF were only produced during the first and second induction. These findings led to specific culture conditions for selective and enhanced production of individual lymphokines.
Assuntos
Linfocinas/metabolismo , Animais , Células Cultivadas , Fatores Estimuladores de Colônias/metabolismo , Concanavalina A/farmacologia , Interleucina-2/biossíntese , Interleucina-5 , Cinética , Fatores Inibidores da Migração de Macrófagos/metabolismo , Camundongos , Oxigênio , Baço/imunologiaRESUMO
The lymphokine interleukin-2 (IL-2) was produced in murine or human lymphocyte cultures by stimulation with concanavalin A (Con A) or phytohemagglutinin (PHA). IL-2 activity in the culture supernatant usually reached an optimum after 20-24 h, followed by a decrease at later times. The duration of IL-2 release was found to be limited by low molecular weight inhibitory substances. These substances reversibly inhibited further IL-2 release at later times. It could be excluded that IL-2 yields in vitro were primarily limited by protease action, inactivation of the mitogen, consumption of nutrients, "product inhibition" by IL-2 or by suppression through cell/cell interaction with hypothetic suppressor cells. It was also excluded that IL-2 activity in the culture supernatant was more quickly consumed by its target cells, the T blasts, than released by its producing cells. To overcome the limitation of IL-2 release by inhibitors, a procedure of repeated induction was developed. Spleen cells or peripheral blood lymphocytes were restimulated several times for 20-24 h with fresh medium and mitogen. This procedure enhanced IL-2 production 2-5 fold compared to a single induction for 24 h.
Assuntos
Interleucina-2/biossíntese , Linfócitos/imunologia , Animais , Concanavalina A/farmacologia , Meios de Cultura , Humanos , Ativação Linfocitária , Camundongos , Fito-Hemaglutininas/farmacologia , Fatores de TempoRESUMO
Cells from different fetal and adult lymphoid organs were tested for the capacity to (a) react to the T-cell growth factor interleukin-2 (IL-2) and (b) to produce IL-2 under appropriate conditions. IL-2 was determined as growth promoting activity for mouse T blasts. Optimal conditions for IL-2 production were: 5 X 10(6) cells/ml; 4-6 micrograms mitogen, 24 h incubation time. Concanavalin A was preferable for fetal thymus, whereas phytohemagglutinin was the appropriate mitogen for lymphocytes from blood and lymph node. Fetal and adult spleen cells produced equal activities of IL-2 irrespective of the mitogen used. Cells from thymus and spleen of 17-21-week-old fetuses produced IL-2 activities like adult cells. Fetal liver cells from the same fetuses produced no IL-2. A comparison of IL-2 activities released from cells of 26 donors showed that within individual variations there was no decrease of the capacity to produce IL-2 with old age. An activity present in culture media of mitogen treated lymphocytes which increased thymidine uptake in fetal liver cells could be distinguished from IL-2 by its different molecular weight and by a heat treatment which abolished IL-2 but not fetal liver cell growth promoting activity. This latter activity is discussed to be colony-stimulating activity. It is concluded that IL-2 is produced by and reacts with T cells only after they have reached or passed the thymus.
Assuntos
Feto/imunologia , Interleucina-2/imunologia , Tecido Linfoide/imunologia , Linfocinas/imunologia , Adulto , Concanavalina A/farmacologia , Substâncias de Crescimento/isolamento & purificação , Humanos , Técnicas In Vitro , Interleucina-2/biossíntese , Interleucina-2/farmacologia , Fígado/imunologia , Ativação Linfocitária , Linfócitos/imunologia , Linfócitos T/imunologiaRESUMO
Human spleen cells or peripheral blood lymphocytes (PBL) were stimulated with the T-cell mitogens concanavalin A or phytohemagglutinin to produce the lymphokine interleukin-2 (IL-2). Culture conditions were optimized using 250-1000 ml reactor vessels. Optimal IL-2 release was found at a low speed of stirring allowing circulation of the medium without suspending the agglutinated cells, and at an oxygen concentration of about 30-45% of saturation. Cell viability was suboptimal at these conditions.
Assuntos
Interleucina-2/biossíntese , Ativação Linfocitária , Linfócitos/fisiologia , Linfocinas/biossíntese , Fito-Hemaglutininas/farmacologia , Células Cultivadas , Concanavalina A/farmacologia , Humanos , Interleucina-2/isolamento & purificação , Linfócitos/efeitos dos fármacos , Baço/fisiologiaRESUMO
Partially purified interleukin 2 (IL-2) promotes proliferation of mouse spleen T, but not B cells, and of peanut-agglutinin-negative (PNA-), and cortisone-resistant "mature" thymocytes, but not of PNA+ "immature" thymocytes. Within the cortisone-resistant thymocyte population, IL-2-responsive cells were found in the blast cell fraction. Proliferation was measured by [3H] thymidine incorporation and subsequent increase in viable cells. The mitogenic effect of IL-2 could not be inhibited by 50 mM methyl-alpha-D-mannoside which excludes contaminating concanavalin A (Con A) as a cause of mitogenicity. The relative increase in viable cells in IL-2 vs. control cultures was abrogated by 1.5 mM hydroxyurea. A possible effect of IL-2 on cell survival is thus ruled out. IL-2, when acting as comitogen with Con A, affected only PNA- and cortisone-resistant thymocytes. These cells also showed high intrinsic IL-2 release when stimulated with Con A such that a comitogenic effect of externally added IL-2 was only seen at low cell concentrations. PNA+ thymocytes could neither be induced to release IL-2 nor did these cells become Con A-responsive under the influence of IL-2, thereby excluding an IL-2-mediated maturation.
Assuntos
Interleucina-2/isolamento & purificação , Linfocinas/isolamento & purificação , Mitógenos/farmacologia , Baço/imunologia , Linfócitos T/classificação , Aglutininas/imunologia , Animais , Arachis , Linfócitos B/imunologia , Separação Celular , Sobrevivência Celular , Interleucina-2/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos CBA , Receptores de Complemento , Glycine max , Linfócitos T/imunologia , Timidina/metabolismoRESUMO
CBA/J mouse splenocytes were induced by concanavalin A to produce the lymphokine costimulator. No serum was needed for this process. Culture conditions were optimized in a 250 ml reactor vessel. While variations of the pH or agitation by stirring had no influence, high dissolved oxygen was found to increase the yields of costimulator even at the cost of cell viability.