Pre-Columbian land use and its modern legacy in the Purus-Madeira Interfluve, Central Amazonia

To combat environmental degradation and change, it is imperative that the rainforests are protected and sustainable land use practices are developed in Amazonia. A better understanding of the role of humans in shaping Amazonian environments and the extent to which the forests have been resilient to anthropogenic disturbance is critical to determining the current state of these ecosystems. This research provides the first reconstruction of late pre-Columbian to early post-Columbian land use and its environmental legacy in the Purus-Madeira Interfluve, Central Amazonia. Soil profile samples were collected across a transect approximately 600 km in length between Manaus and Humaitá, covering a large ecological gradient from dense canopy forests to open canopy forests, as well as dry, upland areas (terra firme) and small riverine settings. Archaeobotanical phytolith and terrestrial palaeoecological samples were analysed from four contexts: (i) primary forests; (ii) oligarchic forests dominated by economically useful trees in the terra firme rainforest on natural soils; (iii) an anthropogenic forest with Brazil nut trees on anthropogenic soil; and (iv) a previously undocumented archaeological site next to the Brazil nut stand. The outcome of this study provides evidence that the extent of the preColumbian environmental impact was larger than previously thought, and this shows that humans managed these forests in various ways to varying intensities. The data, therefore, helps to identify the long-term role of human-environment interactions in Central Amazonia and provides valuable information for future environmental and land use regulation policies

Freeform Bioprinting of Liver Encapsulated in Alginate Hydrogels Tissue Constructs for Pharmacokinetic Study

An in vitro model that can be realistically and inexpensively used to predict human response to various drug administration and toxic chemical exposure is needed. By fabricating a microscale 3D physiological tissue construct consisting of an array of channels and tissue-embedded chambers, one can selectively develop various biomimicking mammalian tissues for a number of pharmaceutical applications, for example, experimental pharmaceutical screening for drug efficacy and toxicity along with apprehending the disposition and metabolic profile of a candidate drug. This paper addresses issues relating to the development and implementation of a bioprinting process for freeform fabrication of a 3D cell-encapsulated hydrogel-based tissue construct, the direct integration onto a microfluidic device for pharmacokinetic study, and the underlying engineering science for the fabrication of a 3D microscale tissue chamber as well as its application in pharmacokinetic study. To this end, a prototype 3D microfluidic tissue chamber embedded with liver cells encapsulated within a hydrogel matrix construct is bioprinted as a physiological in vitro model for pharmacokinetic study. The developed fabrication processes are further validated and parameters optimized by assessing cell viability and liver cell phenotype, in which metabolic and synthetic liver functions are quantitated.Mechanical Engineerin

Hydrofocusing Bioreactor Produces Anti-Cancer Alkaloids

A methodology for growing three-dimensional plant tissue models in a hydrodynamic focusing bioreactor (HFB) has been developed. The methodology is expected to be widely applicable, both on Earth and in outer space, as a means of growing plant cells and aggregates thereof under controlled conditions for diverse purposes, including research on effects of gravitation and other environmental factors upon plant growth and utilization of plant tissue cultures to produce drugs in quantities greater and at costs lower than those of conventional methodologies. The HFB was described in Hydro focus - ing Bioreactor for Three-Dimensional Cell Culture (MSC-22358), NASA Tech Briefs, Vol. 27, No. 3 (March 2003), page 66. To recapitulate: The HFB offers a unique hydrofocusing capability that enables the creation of a low-shear liquid culture environment simultaneously with the herding of suspended cells and tissue assemblies and removal of unwanted air bubbles. The HFB includes a rotating cell-culture vessel with a centrally located sampling port and an internal rotating viscous spinner attached to a rotating base. The vessel and viscous spinner can be made to rotate at the same speed and direction or different speeds and directions to tailor the flow field and the associated hydrodynamic forces in the vessel in order to obtain low-shear suspension of cells and control of the locations of cells and air bubbles. For research and pharmaceutical-production applications, the HFB offers two major benefits: low shear stress, which promotes the assembly of cells into tissue-like three-dimensional constructs; and randomization of gravitational vectors relative to cells, which affects production of medicinal compounds. Presumably, apposition of plant cells in the absence of shear forces promotes cell-cell contacts, cell aggregation, and cell differentiation. Only gentle mixing is necessary for distributing nutrients and oxygen. It has been postulated that inasmuch as cells in the simulated microgravitation of an HFB do not need to maintain the same surface forces as in normal Earth gravitation, they can divert more energy sources to growth and differentiation and, perhaps, to biosynthesis of greater quantities of desired medicinal compounds. Because one can adjust the HFB to vary effective gravitation, one can also test the effects of intermediate levels of gravitation on biosynthesis of various products. The potential utility of this methodology for producing drugs was demonstrated in experiments in which sandalwood and Madagascar periwinkle cells were grown in an HFB. The conditions in the HFB were chosen to induce the cells to form into aggregate cultures that produced anti-cancer indole alkaloids in amounts greater than do comparable numbers of cells of the same species cultured according to previously known methodologies. The observations made in these experiments were interpreted as suggesting that the aggregation of the cells might be responsible for the enhancement of production of alkaloids

Fluid bubble eliminator

A gas-liquid separator uses a helical passageway to impart a spiral motion to a fluid passing therethrough. The centrifugal fore generated by the spiraling motion urges the liquid component of the fluid radially outward which forces the gas component radially inward. The gas component is then filtered through a gas-permeable, liquid-impervious membrane and discharged through a central passageway

Miniature Bioreactor System for Long-Term Cell Culture

A prototype miniature bioreactor system is designed to serve as a laboratory benchtop cell-culturing system that minimizes the need for relatively expensive equipment and reagents and can be operated under computer control, thereby reducing the time and effort required of human investigators and reducing uncertainty in results. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere at a controlled temperature, and associated control subsystems. The system can be used to culture both anchorage-dependent and suspension cells, which can be either prokaryotic or eukaryotic. Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. By integrating this system with one or more microanalytical instrument(s), one can construct a complete automated analytical system that can be tailored to perform one or more of a large variety of assays

Skeletal muscle satellite cells cultured in simulated microgravity

Satellite cells are postnatal myoblasts responsible for providing additional nuclei to growing or regenerating muscle cells. Satellite cells retain the capacity to proliferate and differentiate in vitro and therefore provide a useful model to study postnatal muscle development. Most culture systems used to study postnatal muscle development are limited by the two-dimensional (2-D) confines of the culture dish. Limiting proliferation and differentiation of satellite cells in 2-D could potentially limit cell-cell contacts important for developing the level of organization in skeletal muscle obtained in vivo. Culturing satellite cells on microcarrier beads suspended in the High-Aspect-Ratio-Vessel (HARV) designed by NASA provides a low shear, three-dimensional (3-D) environment to study muscle development. Primary cultures established from anterior tibialis muscles of growing rats (approximately 200 gm) were used for all studies and were composed of greater than 75 % satellite cells. Different inoculation densities did not affect the proliferative potential of satellite cells in the HARV. Plating efficiency, proliferation, and glucose utilization were compared between 2-D flat culture and 3-D HARV culture. Plating efficiency (cells attached - cells plated x 100) was similar between the two culture systems. Proliferation was reduced in HARV cultures and this reduction was apparent for both satellite cells and non-satellite cells. Furthermore, reduction in proliferation within the HARV could not be attributed to reduced substrate availability since glucose levels in media from HARV and 2-D cell culture were similar. Morphologically, microcarrier beads within the HARVS were joined together by cells into three-dimensional aggregates composed of greater than 10 beads/aggregate. Aggregation of beads did not occur in the absence of cells. Myotubes were often seen on individual beads or spanning the surface of two beads. In summary, proliferation and differentiation of satellite cells on microcarrier beads within the HARV bioreactor results in a three dimensional level of organization that could provide a more suitable model to study postnatal muscle development