Higher production of rabies virus in serum-free medium cell cultures on microcarriers.

Rabies virus suspensions were obtained from VERO cells cultivated on solid microcarriers in a bioreactor after infection with the Pasteur rabies virus strain (PV). Virus production-serum free medium (VP-SFM) or Leibovitz 15 (L15) medium supplemented or not with fetal calf serum (FCS) were used to cultivate the VERO cells, before and after virus infection. The cell growth was shown to reach higher densities (1.6 x 10(6) cellsmol(-l)), when VP-SFM supplemented with 1% of FCS was used during the cell growth phase of culture, and then replaced by VP-SFM alone for the virus multiplication phase. In the cultures performed from the beginning with VP-SFM, lower densities accompanied by an altered cell morphology and detachment from the microcarriers were always observed. In rabies virus infected cultures, kinetic studies showed that higher virus yields (10(4.7) FFD(50) per 0.05 ml) were always obtained in cultures performed initially on VP-SFM supplemented with 1% FCS and after infection on VP-SFM alone. In agreement with that, rabies virus production, as measured by the average of virus titers in harvests obtained at different times after infection were shown to be 5.5 times higher in the cell cultures using initially VP-SFM+1%FCS and, following infection, VP-SFM alone. Besides the advantages of using media with a well-controlled composition, these data indicate the usefulness of serum free media also in terms of virus productivity.

Long-term accumulation and microdistribution of uranium in the bone and marrow of beagle dog.

The accumulation and microdistribution of uranium in the bone and marrow of Beagle dogs were determined by both neutron activation and neutron-fission analysis. The experiment started immediately after the weaning period, lasting till maturity. Two animal groups were fed daily with uranyl nitrate at concentrations of 20 and 100 microg g(-1) food. Of the two measuring techniques, uranium accumulated along the marrow as much as in the bone, contrary to the results obtained with single, acute doses. The role played by this finding for the evaluation of radiobiological long-term risks is discussed. It was demonstrated, by means of a biokinetical approach, that the long-term accumulation of uranium in bone and marrow could be described by a piling up of single dose daily incorporation.

Evaluation of a program to reduce back pain in nursing personnel.

OBJECTIVE: To evaluate the effectiveness of a program designed to reduce back pain in nursing aides. METHODS: Female nursing aides from a university hospital who had suffered episodes of back pain for at least six months were included in the study. Participants were randomly divided into a control group and an intervention group. The intervention program involved a set of exercises and an educational component stressing the ergonomic aspect, administered twice a week during working hours for four months. All subjects answered a structured questionnaire and the intensity of pain was assessed before and after the program using a visual analogue scale (VAS). Student's t-test or the Wilcoxon Rank Sum Test for independent samples, and Chi-square test or the Exact Fisher test for categorical analysis, were used. The McNemar test and the Wilcoxon matched pairs test were used to compare the periods before and after the program. RESULTS: There was a statistically significant decrease in the frequency of cervical pain in the last two months and in the last seven days in the intervention group. There was also a reduction in cervical pain intensity in the two periods (2 months, 7 days) and lumbar pain intensity in the last 7 days. CONCLUSIONS: The results suggest that a program of regular exercise with an emphasis on ergonomics can reduce musculoskeletal symptoms in nursing personnel.

[Invasive procedures and complications in patients with craniocerebral trauma hospitalized in an intensive therapy unit]. / Procedimentos invasivos e complicações em pacientes com traumatismo crâneo-encefálico internados em unidade de terapia intensiva.

The present study realized with 50 patients from an Intensive Care Unit, due to head injury and considering that the majority of them presented a score equal or below six (6) in the Glasgow Coma Scale, has shown that the utilization of invasive procedures is high. Regarding complications, there was predominance of fever, upper airway secretions, hematomas at the arterial puncture localization and diarrhea. Proportionally, the patients under a clinical treatment presented a lower number of invasive procedures and problems and the reverse occurred with those under surgery. The analysis of the association of the invasive procedures and problems compared to the level of consciousness has been affected.

[Health and education: alternative courses for the development of nursing personnel]. / Saúde e educação: cursos alternativos para desenvolvimento do pessoal de enfermagem.

To promote therapeutic educational activities for nursing personnel in order to decrease stress, to improve interpersonal relations and the search for self-knowledge are the objectives of the courses promoted by the Departament of Nursing of FCM and by The Continued Education Nursing Service of the University Hospital of UNICAMP. Respiration, relaxation, body sensibilization and awareness, and theater interpretation techniques were taught in the following courses: The Hospital and Human Relations, Dance and Creativity, Yoga and Mental Relaxation. The estrategy employed was "group experience", with the participation of nurse's aides, nurses technicians, practical nurses, and registered nurses during working hours in 15 to 20 meetings per course. "Individual statements" written by the participants were used as a research tool, and the method employed was content analysis. The evaluation demonstrated that the "experience" facilitated relations among the members of the work team, opened the space for effective communication, favored self-knowledge, and helped with the problem solving. The analysis demonstrated the importance of the continuity of alternative courses to help employes improve their relations with themselves, with others and with their work, and to properly value their health/learning.

Kinetic studies of recombinant rabies virus glycoprotein (RVGP) cDNA transcription and mRNA translation in Drosophila melanogaster S2 cell populations.

Recombinant rabies virus glycoprotein (RVGP) was expressed in cell membranes of stably transfected Drosophila S2 cells using constitutive and inducible promoters. Although with quantitative differences of RVGP expression in both systems, the cDNA transcription, as evaluated by relative RVGP mRNA levels measured by qRT-PCR, sustained the amount of RVGP producing cells and the RVGP volumetric (ΠRVGP) productivity. At the transition to the stationary cell growth phase, once the cell culture slowed down its rate of multiplication, an accumulation of RVGP mRNA and RVGP was clearly observed in both cell populations. Nevertheless, cell cultures performed under sub-optimal temperatures indicated that an envisaged increase in the RVGP production is not only dependent on cell growth rate, but essentially on optimal cell metabolic state.

Kinetic response of a Drosophila melanogaster cell line to different medium formulations and culture conditions.

In the past few years, Drosophila melanogaster cells have been employed for recombinant protein production purposes, and a comprehensive knowledge of their metabolism is essential for process optimization. In this work, the kinetic response of a Schneider S2 cell line, grown in shake flasks, in two different culture media, the serum-free SF900-II((R)) and the serum-supplemented TC-100, was evaluated. Cell growth, amino acids and glucose uptake, and lactate synthesis were measured allowing the calculation of kinetic parameters. The results show that S2 cells metabolism was able to adjust to different environmental situations, as determined by medium formulation, as well as by the particular situation resulting from the culture conditions. Cells attained a 163% higher final cell concentration (1.4 x 10(7) cells mL(-1)) in SF900 II((R)) medium, when compared to serum-supplemented TC-100 medium. Also, a maximum specific cell growth rate 52% higher in SF900 II((R) )medium, when compared to serum-supplemented TC-100 one, was observed. Glutamine was the growth limiting factor in SF900 II((R)) medium, while glucose, sometimes associated with glutamine, controlled growth in serum-supplemented TC-100 medium based formulation. The different pattern of lactate production is an example of the versatility of the metabolism of these cells. This by-product was produced only in glutamine limitation, but the amount synthesized depended not only on the excess glucose, but on other medium components. Therefore, in serum-supplemented TC-100 medium a much smaller lactate amount was generated. Besides, glucose was identified not only as a growth limiting factor, but also as a viability limiting factor, since its depletion accelerated cell death.

Nó-de-cachorro (Heteropterys tomentosa A. Juss.): espécie de uso medicinal em Mato Grosso, Brasil / Nó-de-cachorro (Heteropterys tomentosa A. Juss.): a medicinal plant from Mato Grosso State, Brazil

Neste estudo é apresentada uma revisão sobre nó-de-cachorro (Heteropterys tomentosa A. Juss.), espécie de uso medicinal em Mato Grosso. São abordadas as características da espécie com a descrição da planta, hábitat, distribuição geográfica, ecologia, fenologia e biologia reprodutiva, propriedades químicas e farmacológicas. Os aspectos como a etnobotânica (uso e partes usadas), formas de exploração atual e informações sobre cultivo são discutidos.

This study presents a review about "Nó-de-cachorro" (Heteropterys tomentosa A. Juss.), a medicinal species from Mato Grosso State, Brazil. We outlined the characteristics of the species including the plant description, habitat, geographic distribution, ecology, phenology and reproductive biology, chemical and pharmacological properties. Aspects such as ethnobotany (use and used parts), current ways of exploiting and information on cultivation are discussed.

Rabies virus production in high vero cell density cultures on macroporous microcarriers.

The purpose of the study was to investigate the rabies virus multiplication in Vero cell cultures performed on porous microcarriers, MCs (cellulose-Cytopore and gelatin-Cultispher G), which provide higher available surface area compared with solid (nonporous) MCs (DEAE-Cytodex 1). In a set of experiments performed at the same MC concentration (MCs per milliliter), cell densities regularly obtained in porous MC cultures were comparable, but almost twice as high as those in solid MC cultures. In addition, 41.1 +/- 3.9-, 35.2 +/- 2-, and 19.6 +/- 5.8-fold increases in cell concentration, relative to the initial cell number, along with maximum rabies virus titers of 6.3 +/- 0.3 x 10(4), 5 +/- 0.1 x 10(4), and 4.3 +/- 0.2 x 10(4) FFD(50)/mL were observed in Cytopore, Cultispher G, and Cytodex 1 MC cultures, respectively. When higher concentrations of MCs were employed, lower performances of virus production and MC-cell occupation (cells per MC or cells per square millimeter) were observed. Cell attachment to MCs was shown to be faster for Cytopore MCs and Cytodex 1 MCs than for Cultispher G MCs. Concerning the kinetics of cell multiplication on MCs, exponential cell growth, at similar specific cell growth rates, took place on Cytopore, Cultispher G, and Cytodex 1 MCs. In addition, cell densities as high as 2.1 +/- 0.2 x 10(6) cells/mL on Cytopore MCs, 1.8 +/- 0.1 x 10(6) cells/mL on Cultispher G MCs, and 1 +/- 0.3 x 10(6) cells/mL on Cytodex 1 MCs were regularly obtained in batch cultures. Optical as well as scanning and transmission electron microscopy studies carried out to analyze MC structure, MC cell occupation, and cell permissivity to virus infection demonstrated that there was uniform cell distribution in the external and internal areas of the MCs, suggesting an efficiency of virus synthesis. Our results indicate the usefulness of these supports for rabies virus antigen production, as well as possibilities for further optimization.

Expression of the hepatitis B virus surface antigen in mammalian cells using an Epstein-barr-virus-derived vector.

The hepatitis B virus surface antigen (HBsAg) gene, under control of the inducible mouse metallothionein I gene promoter, was inserted in an expression vector based on the Epstein-Barr virus (EBV). This vector was introduced into human cells by DNA transfection and clones were selected for their resistance to hygromycin B. The recombinant EBV vector replicates efficiently as an episome in human cells and approximately six copies per cell were found in one clone of hygromycin-B-resistant cells. These cells produce high levels of HBsAg in the presence of metals. The protein is mainly found in the cell medium, suggesting that the HBsAg is secreted from the cells.