Successful scale-up of human embryonic stem cell production in a stirred microcarrier culture system

Future clinical applications of human embryonic stem (hES) cells will require high-yield culture protocols. Currently, hES cells are mainly cultured in static tissue plates, which offer a limited surface and require repeated sub-culturing. Here we describe a stirred system with commercial dextran-based microcarriers coated with denatured collagen to scale-up hES cell production. Maintenance of pluripotency in the microcarrier-based stirred system was shown by immunocytochemical and flow cytometry analyses for pluripotency-associated markers. The formation of cavitated embryoid bodies expressing markers of endoderm, ectoderm and mesoderm was further evidence of maintenance of differentiation capability. Cell yield per volume of medium spent was more than 2-fold higher than in static plates, resulting in a significant decrease in cultivation costs. A total of 10(8) karyotypically stable hES cells were obtained from a unitary small vessel that needed virtually no manipulation during cell proliferation, decreasing risks of contamination. Spinner flasks are available up to working volumes in the range of several liters. If desired, samples from the homogenous suspension can be withdrawn to allow process validation needed in the last expansion steps prior to transplantation. Especially when thinking about clinical trials involving from dozens to hundreds of patients, the use of a small number of larger spinners instead of hundreds of plates or flasks will be beneficial. To our knowledge, this is the first description of successful scale-up of feeder- and Matrigel™-free production of undifferentiated hES cells under continuous agitation, which makes this system a promising alternative for both therapy and research needs.

Desenvolvimento de tecnologia para confecção de eletrodos e conjuntos eletrodo-membrana-eletrodo (MEA) por impressão à tela para aplicação em módulos de potência de células PEMFC

Significant functions in the Proton Exchange Membrane Fuel Cells (PEMFCs) rely on Gas Diffusion Layers (GDLs), such as control the water balance in the membrane electrode assembly (MEA), allow suitable gas permeability and porosity, etc. Aware of the GDL importance in the cell performance and its great demand in scale-up projects, the fuel cell research group at Instituto de Pesquisas Energéticas e Nucleares (IPEN) has developed a Sieve Printing method (innovative in Brazil) as a strategic solution for producing GDL and electrodes used in high power PEMFC stacks. The method has shown to be adequate to fabricate low cost electrodes, GDLs of different dimensions and to produce any amount of MEAs for power stacks.

Payment for performance in the Family Health Programme: lessons from the UK Quality and Outcomes Framework

OBJECTIVE: Payment for performance financial incentive schemes reward doctors based on the quality and the outcomes of their treatment. In Brazil, the Ministry of Health is looking to scale up its use in public hospitals and some municipalities are developing payment for performance schemes even for the Family Health Programme. In this article the Quality and Outcomes Framework used in the UK since 2004 is discussed, as well as its experience to elaborate some important lessons that Brazilian municipalities should consider before embarking on payment for performance scheme in primary care settings.

r-Sm14 - pRSETA efficacy in experimental animals

Previous studies carried out with Sm14 in experimental vaccination against Schistosoma mansoni or Fasciola hepatica infections were performed with recombinant Sm14 (rSm14) produced in Escherichia coli by the pGEMEX system (Promega). The rSm14 was expressed as a 40 kDa fusion protein with the major bacteriophage T7 capsid protein. Vaccination experiments with this rSm14 in animal models resulted in consistent high protective activity against S. mansoni cercariae challenge and enabled rSm14 to be included among the vaccine antigens endorsed by the World Health Organization for phase I/II clinical trials. Since the preparation of pGEMEX based rSm14 is time consuming and results in low yield for large scale production, we have tested other E. coli expression systems which would be more suitable for scale up and downstream processing. We expressed two different 6XHis-tagged Sm14 fusion proteins in a T7 promoter based plasmids. The 6XHis-tag fusions allowed rapid purification of the recombinant proteins through a Ni+2-charged resin. The resulted recombinant 18 and 16 kDa proteins were recognized by anti-Sm14 antibodies and also by antiserum against adult S. mansoni soluble secreted/excreted proteins in Western-Blot. Both proteins were also protective against S. mansoni cercariae infection to the same extent as the rSm14 expressed by the pGEMEX system.

Efeito da adição de precursores na produção de alcaloide anticancerígeno usando a técnica de planejamento experimental

The effect of precursors on the anticancer alkaloid production by submerged fermentation using M. anisopliae 3935 was studied, according to complete experimental design 2² with three central points. The results showed that lysine was the most important variable, however, when both lysine and glucose were added to the fermentation medium, the alkaloid production reached, approximately, 17 mg L-1 after 120 hours of fermentation. Then, the scale-up of the process was carried out and these results were confirmed. Finally, 35 mg L-1 of alkaloid at 192 h were attained after increment of added aminoacid lysine.

Nanopartículas de lipídios sólidos: métodos clássicos de produção laboratorial

Solid lipid nanoparticles have been extensively investigated as drug delivery systems. These colloidal systems have major advantages compared to others more traditional. Reported advantages include sustained release, ability to solubilize lipophilic drugs, increased physical and chemical stability of labile molecules, decreased unwanted side-effects showing lower toxicity, and scale up facilities. This paper aims at reviewing the traditional methods of solid lipid nanoparticles production, such as fusion-emulsification (hot and cold), solvent evaporation-emulsification and microemulsion, dealing with the main technological parameters that influence the quality properties of solid lipid nanoparticles.

Processo de transferência de tecnologia da universidade para a indústria: estudo de caso envolvendo a conversão de glicerol

Currently, public policy has encouraged innovation in universities and also transference of technology to the industry. Another important stage to be considered would be the registration or filing of a patent and the economical viability study. Government programs, such as the innovation incentive program, among others, should facilitate popularization and promote interest by industry. In this work we described the steps, from the conception of the idea to the scale up going through its interest by the industry. The case study is about the glycerol conversion utilizing modified niobia as catalysts.

A utilização de etanol em célula a combustível de óxido sólido

This work reports a review on the status and technical feasibility of the application of ethanol as fuel for Solid Oxide Fuel Cells (SOFC), presenting both external reform and cell with direct utilization of ethanol. Based on this survey, both experimental results and mathematical modeling indicated the technical feasibility of power generation by ethanol SOFC, with cell units producing 450 mW/cm², sufficient for scale up to large stationary plants. The quantitative assessments in the literature show this field to be promising for researchers and private sector investment as well being a strategic technology for government policy in the short and long term.

Impactos da nanotecnologia na saúde: produção de medicamentos

Several studies have described the benefits of nanoscience and nanotechnology (N&N) in different sectors such as agriculture, energy, environmental preservation, and public health. The rapid evolution of N&N can be shown through a panoramic analysis of scientific papers and patents. In the area of public health, it is estimated that the global market for nanotechnology products will expand to 160 billion U.S. dollars in 2015. The Brazilian government has also strengthened its innovative potential in N&N through economic subsidies, as observed for other countries. This review is focused on the current landscape of N&N in a therapeutic context, highlighting the development of nanotech-products produced with biocompatible and biodegradable materials that are already commercially available or under investigation. Most studies under investigation are focused on the development of nanotechnology-based formulations intended for treatment of cancer, inflammatory, cardiovascular, and neurological diseases. Although there are several advantages of N&N in healthcare, many challenges have to be conquered to increase the availability of nanotechnology products in toxicological, preclinical and clinical studies, scale-up, regulatory, and private investments.

Química Sem Fronteiras: o desafio da energia

Coal, natural gas and petroleum-based liquid fuels are still the most widely used energy sources in modern society. The current scenario contrasts with the foreseen shortage of petroleum that was spread out in the beginning of the XXI century, when the concept of "energy security" emerged as an urgent agenda to ensure a good balance between energy supply and demand. Much beyond protecting refineries and oil ducts from terrorist attacks, these issues soon developed to a portfolio of measures related to process sustainability, involving at least three fundamental dimensions: (a) the need for technological breakthroughs to improve energy production worldwide; (b) the improvement of energy efficiency in all sectors of modern society; and (c) the increase of the social perception that education is a key-word towards a better use of our energy resources. Together with these technological, economic or social issues, "energy security" is also strongly influenced by environmental issues involving greenhouse gas emissions, loss of biodiversity in environmentally sensitive areas, pollution and poor solid waste management. For these and other reasons, the implementation of more sustainable practices in our currently available industrial facilities and the search for alternative energy sources that could partly replace the fossil fuels became a major priority throughout the world. Regarding fossil fuels, the main technological bottlenecks are related to the exploitation of less accessible petroleum resources such as those in the pre-salt layer, ranging from the proper characterization of these deep-water oil reservoirs, the development of lighter and more efficient equipment for both exploration and exploitation, the optimization of the drilling techniques, the achievement of further improvements in production yields and the establishment of specialized training programs for the technical staff. The production of natural gas from shale is also emerging in several countries but its production in large scale has several problems ranging from the unavoidable environmental impact of shale mining as well as to the bad consequences of its large scale exploitation in the past. The large scale use of coal has similar environmental problems, which are aggravated by difficulties in its proper characterization. Also, the mitigation of harmful gases and particulate matter that are released as a result of combustion is still depending on the development of new gas cleaning technologies including more efficient catalysts to improve its emission profile. On the other hand, biofuels are still struggling to fulfill their role in reducing our high dependence on fossil fuels. Fatty acid alkyl esters (biodiesel) from vegetable oils and ethanol from cane sucrose and corn starch are mature technologies whose market share is partially limited by the availability of their raw materials. For this reason, there has been a great effort to develop "second-generation" technologies to produce methanol, ethanol, butanol, biodiesel, biogas (methane), bio-oils, syngas and synthetic fuels from lower grade renewable feedstocks such as lignocellulosic materials whose consumption would not interfere with the rather sensitive issues of food security. Advanced fermentation processes are envisaged as "third generation" technologies and these are primarily linked to the use of algae feedstocks as well as other organisms that could produce biofuels or simply provide microbial biomass for the processes listed above. Due to the complexity and cost of their production chain, "third generation" technologies usually aim at high value added biofuels such as biojet fuel, biohydrogen and hydrocarbons with a fuel performance similar to diesel or gasoline, situations in which the use of genetically modified organisms is usually required. In general, the main challenges in this field could be summarized as follows: (a) the need for prospecting alternative sources of biomass that are not linked to the food chain; (b) the intensive use of green chemistry principles in our current industrial activities; (c) the development of mature technologies for the production of second and third generation biofuels; (d) the development of safe bioprocesses that are based on environmentally benign microorganisms; (e) the scale-up of potential technologies to a suitable demonstration scale; and (f) the full understanding of the technological and environmental implications of the food vs. fuel debate. On the basis of these, the main objective of this article is to stimulate the discussion and help the decision making regarding "energy security" issues and their challenges for modern society, in such a way to encourage the participation of the Brazilian Chemistry community in the design of a road map for a safer, sustainable and prosper future for our nation.

SEPARATION OF SATURED AND UNSATURATED FATTY ACIDS FROM PALM FATTY ACIDS DISTILLATES IN CONTINUOUS MULTISTAGE COUNTERCURRENT COLUMNS WITH SUPERCRITICAL CARBON DIOXIDE AS SOLVENT: A PROCESS DESIGN METHODOLOGY

In this work the separation of multicomponent mixtures in counter-current columns with supercritical carbon dioxide has been investigated using a process design methodology. First the separation task must be defined, then phase equilibria experiments are carried out, and the data obtained are correlated with thermodynamic models or empirical functions. Mutual solubilities, Ki-values, and separation factors aij are determined. Based on this data possible operating conditions for further extraction experiments can be determined. Separation analysis using graphical methods are performed to optimize the process parameters. Hydrodynamic experiments are carried out to determine the flow capacity diagram. Extraction experiments in laboratory scale are planned and carried out in order to determine HETP values, to validate the simulation results, and to provide new materials for additional phase equilibria experiments, needed to determine the dependence of separation factors on concetration. Numerical simulation of the separation process and auxiliary systems is carried out to optimize the number of stages, solvent-to-feed ratio, product purity, yield, and energy consumption. Scale-up and cost analysis close the process design. The separation of palmitic acid and (oleic+linoleic) acids from PFAD-Palm Fatty Acids Distillates was used as a case study.

Scientific authorships and collaboration network analysis on Chagas disease: papers indexed in PubMed (1940-2009)

Chagas disease is a chronic, tropical, parasitic disease, endemic throughout Latin America. The large-scale migration of populations has increased the geographic distribution of the disease and cases have been observed in many other countries around the world. To strengthen the critical mass of knowledge generated in different countries, it is essential to promote cooperative and translational research initiatives. We analyzed authorship of scientific documents on Chagas disease indexed in the Medline database from 1940 to 2009. Bibliometrics was used to analyze the evolution of collaboration patterns. A Social Network Analysis was carried out to identify the main research groups in the area by applying clustering methods. We then analyzed 13,989 papers produced by 21,350 authors. Collaboration among authors dramatically increased over the study period, reaching an average of 6.2 authors per paper in the last five-year period. Applying a threshold of collaboration of five or more papers signed in co-authorship, we identified 148 consolidated research groups made up of 1,750 authors. The Chagas disease network identified constitutes a "small world," characterized by a high degree of clustering and a notably high number of Brazilian researchers.

Breeding of Biomphalaria tenagophila in mass scale

An efficient method for breeding Biomphalaria tenagophila (Taim lineage/RS) was developed over a 5-year-period (2005-2010). Special facilities were provided which consisted of four cement tanks (9.4 x 0.6 x 0.22 m), with their bottom covered with a layer of sterilized red earth and calcium carbonate. Standard measures were adopted, as follows: each tank should contain an average of 3000 specimens, and would be provided with a daily ration of 35,000 mg complemented with lettuce. A green-house effect heating system was developed which constituted of movable dark canvas covers, which allowed the temperature to be controlled between 20 - 24 ºC. This system was essential, especially during the coldest months of the year. Approximately 27,000 specimens with a diameter of 12 mm or more were produced during a 14-month-period. The mortality rates of the newly-hatched and adult snails were 77% and 37%, respectively. The follow-up of the development system related to 310 specimens of B. tenagophila demonstrated that 70-day-old snails reached an average of 17.0 ± 0.9 mm diameter. The mortality rates and the development performance of B. tenagophila snails can be considered as highly satisfactory, when compared with other results in literature related to works carried out with different species of the genus Biomphalaria, under controlled laboratory conditions.

A five year follow-up study of 11 patients with bipolar disorder

OBJECTIVE: To study the long-term follow-up of patients with bipolar disorder (BPD). METHOD: Eleven outpatients with BPD type I were followed up naturalistically for five years at a university teaching hospital. The Clinical Global Impression Scale (BPD version) was used to evaluate the occurrence of affective episodes, and the Strauss-Carpenter Outcome Scale was used to evaluate social and occupational functioning. RESULTS: The majority of patients were symptomatic most of the time, with predominantly depressive episodes. Overall, patients remained euthymic a mean of 47.7% of the time. Despite a low rate of hospitalization, social and occupational functioning was poor in the majority of patients. A poor disease course with respect to work-related functioning was associated with fewer months of euthymia with a longer duration of depressive episodes. The total number of months of euthymia negatively correlated with the patient's age and disease duration. CONCLUSION: Despite the small sample size, the present findings appear to corroborate previous studies on the evolution of BPD. Most of the patients had a poor disease course, with long symptomatic periods, particularly depressive episodes, and significantly impaired social and occupational functioning.

Set-up of a method using LC-UV to assay mometasone furoate in pharmaceutical dosage forms

A reliable method using LC-UV to assay mometasone furoate (MF) in creams or nasal sprays using the same chromatographic conditions was set up. Methanol:water 80:20 (v/v) (1.0 mL min-1) was used as mobile phase. MF was detected at 248 nm and analyzed in a concentration range from 1.0 to 20.0 µg mL-1. The method provided acceptable theoretical plates, peak simmetry, peak tailing factor and peak resolution a short run (5 min). The method showed specificity, good linearity (r = 0.9999) and the quantification limit was 0.379 µg mL-1. Furthermore, the method was precise (RSD < 2.0%), accurate (recovery > 97%) and robust.