Inhibition of Phagocytosis-Associated Chemiluminescence by Superoxide Dismutase

During the process of phagocytosis, human leukocytes emit a burst of luminescence which can be measured in a liquid scintillation spectrometer. The enzyme superoxide dismutase, which removes superoxide anions (O(2.)), inhibited this chemiluminescence by 70% at a concentration of 100 mug/ml. The enzyme did not inhibit phagocytosis. These results support other studies indicating that O(2.) is elaborated by phagocytizing leukocytes. They also indicate that O(2.) plays a major role in phagocytosis-associated chemiluminescence, though not necessarily as the luminescing agent. Catalase and benzoate inhibited the chemiluminescence of phagocytosis to a slight extent, suggesting that hydrogen peroxide and hydroxyl radical, respectively, might also be involved in this phenomenon. The relationship between the mediators of chemiluminescence and those responsible for phagocytic bactericidal activity remains to be defined.     

Galaxy: a platform for interactive large-scale genome analysis

Accessing and analyzing the exponentially expanding genomic sequence and functional data pose a challenge for biomedical researchers. Here we describe an interactive system, Galaxy, that combines the power of existing genome annotation databases with a simple Web portal to enable users to search remote resources, combine data from independent queries, and visualize the results. The heart of Galaxy is a flexible history system that stores the queries from each user; performs operations such as intersections, unions, and subtractions; and links to other computational tools. Galaxy can be accessed at http://g2.bx.psu.edu.     

Composite articular cartilage engineered on a chondrocyte-seeded aliphatic polyurethane sponge

To circumvent the reconstructive disadvantages inherent in resorbable polyglycolic acid (PGA)/polylactic acid (PLA) used in cartilage engineering, a nonresorbable, and nonreactive polyurethane sponge (Tecoflex sponge, TS) was studied as both a cell delivery device and as an internal support scaffolding. The in vitro viability and proliferation of porcine articular chondrocytes (PACs) in TS, and the in vivo generation of new articular cartilage and long-term resorption, were examined. The initial cell attachment rate was 40%, and cell density increased more than 5-fold after 12 days of culture in vitro. PAC-loaded TS blocks were implanted into nude mice, became opalescent, and resembled native cartilage at weeks 12 and 24 postimplantation. The mass and volume of newly formed cartilage were not significantly different at week 24 from samples harvested at week 6 or week 12. Safranin O-fast green staining revealed that the specimens from cell-loaded TS groups at week 12 and week 24 consisted of mature cartilage. Collagen typing revealed that type II collagen was present in all groups of tissue-engineered cartilage. In conclusion, the implantation of PAC-TS resulted in composite tissue-engineered articular cartilage with TS as an internal support. Long-term observation (24 weeks) of mass and volume showed no evidence of resorption.     

Non-cholinergic effects of acetylcholinesterase in the substantia nigra: a possible role for an ATP-sensitive potassium channel

Summary Within the substantia nigra acetylcholinesterase has non-cholinergic actions that can be demonstrated at both behavioural and cellular levels: the aim of this study was, thus, to explore, in the in vitro guinea pig substantia nigra, the ionic mechanisms which mediate these non-classical phenomena. Acetylcholinesterase had a reversible hyperpolarizing action, via an opening of potassium channels, on a selective population of nigral neurons. These neurons could be identified by an ability to generate bursts of action potentials and by a sensitivity to either amphetamine or to a reduction of glucose in the perfusing medium. The acetylcholinesterase-induced hyperpolarization could not be attributed to a contaminant in the exogenous solution, since a highly purified preparation was even more potent. Furthermore, enzymatic action of any kind could be eliminated as boiled acetylcholinesterase was equally efficacious. The effect of acetylcholinesterase was not subject to tachyphylaxis and was resistant to blockade of potassium channels with tetraethylammonium: since both these phenomena are features of the D₂ autoreceptor for dopamine within the substantia nigra, it seems unlikely that acetylcholinesterase is operating on the same target as dendritically released local dopamine. On the other hand, the actions of acetylcholinesterase were enhanced by low glucose and blocked by the sulfonylurea, tolbutamide. These results strongly suggest that acetylcholinesterase can exert a nonenzymatic action and that this action, in the substantia nigra, is mediated by an ATP-sensitive potassium channel.     

Generating compensation designs for tangential breast irradiation with artificial neural networks

In this paper we discuss a study comparing an algorithm implemented clinically to design intensity-modulated fields with two artificial neural networks (ANNs) trained to design the same fields. The purpose of the algorithm is to produce compensation for tangential breast radiotherapy in order to improve dose homogeneity. This was achieved by creating intensity-modulated fields to supplement standard wedged fields. Portal image data were used to create thickness maps of the medial and lateral fields, which in turn were used to design the wedged and intensity-modulated fields. The ANNs were developed to design the intensity-modulated fields from the portal image data and corresponding fluence map alone. One used localized groups of portal image pixels related to the fluence map (method 2), and the other used a one-to-one mapping between spatially corresponding pixels (method 3). A dosimetric comparison of the methods was performed by calculating the overall dose distribution. The volume of tissue outside the dose range 95-105% was used to assess dose homogeneity. The average volume outside 95-105%, averaged over 80 cases, was shown to be 2.3% for the algorithm, whilst average values of 9.9% and 13.5% were obtained for methods 2 and 3, respectively. The results of this study demonstrate the ability of an ANN to learn the general shape of compensation required and explore the use of image-based ANNs in the design of intensity-modulated fields.     

An optimization algorithm that incorporates IMRT delivery constraints

An intensity-modulated beam optimization algorithm is presented which incorporates the delivery constraints into the optimization cycle. The optimization algorithm is based on the quasi-Newton method of iteratively solving minimization problems. The developed algorithm iteratively corrects the incident, pencil-beam-like, fluence to incorporate the delivery constraints. In the present study, the goal of the optimization algorithm is to achieve the best deliverable radiotherapy plan, subject to the constraints of the delivery technique described by a leaf-sequencing algorithm being applied concurrently. In general, if they are applied after, rather than during, the optimization cycle, the delivery constraints associated with the IMRT technique can produce local variations up to 6% in the 'optimized' dose (i.e., distribution without applied constraints) and reduce the degree of conformity, of the dose, to the PTV region. The optimization method has been applied to three IMRT delivery techniques: dynamic multileaf (DMLC), multiple-static-field (MSF) and slice-by-slice tomotherapy (NOMOS MIMiC). The beam profiles were generated for a prostate tumour with organs at risk being the rectum, bladder and femoral heads. The optimization method described was shown to generate optimum and deliverable IMRT plans for these three delivery techniques. In the case of the DMLC and MSF the optimization converged within 3-5 iterations to a mean PTV dose of 69.60 +/- 1.34 Gy and 69.71 +/- 1.34 Gy, respectively, while for NOMOS MIMiC approximately 10 iterations were needed to obtain 69.68 +/- 1.55 Gy. In addition to this, the IMRT optimization also yielded optimum fluence profiles when clustering was performed concurrently with the leaf-sequencer. An optimum between 8 and 15 clusters of equal fluence 'intensity' was shown to establish the best compromise between the number of fluence levels and the PTV dose coverage.     

Concepts for shuttling multileaf collimators for intensity-modulated radiation therapy

Shuttling multileaf collimators (SMLCs) can increase the MU efficiency of intensity-modulated radiation therapy compared with the multiple-static-field (MSF-MLC) technique or dynamic MLC (DMLC) technique with conventional MLCs. In a previous paper (Phys. Med. Biol. 45 3343-58) a particular SMLC was shown, for highly modulated intensity distributions, to increase the MU efficiency compared with the MSF-MLC technique. In this companion paper, two new arrangements similar to that described in the earlier paper, but with less mechanical complexity, are shown to be constructionally simpler but less MU efficient. Additionally another new concept of SMLC is shown which also increases the MU efficiency compared with the MSF-MLC technique and often improves the MU efficiency compared with the previously reported SMLC for highly modulated intensity distributions. It also leads to zero tongue-and-groove underdose in the direction orthogonal to that of the shuttling elements (so-called across-the-rows).     

Assessing lymphocyte functions in neonates for revealing abnormal prenatal development of the immune system

Because it is difficult to assess prenatally induced functional deficits of the human immune system, we developed an ex vivo method for differentiation and maturation of peripheral lymphocytes of newborn, preferentially using umbilical cord blood. Many lymphocyte subsets of newborn infants are "immature" with respect to defined surface receptors. An example of such an immaturity is the almost complete lack of "memory"-type helper T cells (also designated as helper-inducer cells), characterized by expressing the surface receptors: CD4(+)CD45R0(+)CD45RA(-)CD29(high). On the other hand, umbilical cord blood contains many "naive"-type helper T cells (often designated as suppressor-inducer cells), with the receptors: CD4(+)CD45R0(-)CD45RA(+)CD29(low). In this report, we demonstrate that the immature helper lymphocyte population of umbilical cord blood is capable of differentiating to mature cells following stimulation with pokeweed mitogen (PWM) and other stimulants ex vivo. The obtained receptor pattern is virtually indistinguishable from the one observed on the mature cells of adults. Such an extensive differentiation can only be achieved with cells of newborns. As intermediates during differentiation in culture, CD45R0(+)CD45RA(+) cells may be observed which are rather rare in vivo. Additionally, the appearance of several activation (CD25, CD69, HLA-DR) and adhesion (CD11a, CD11b, CD11c, CD18, CD49b, CD49d, CD54) receptors on CD4 cells were analyzed. With this model system evidence for the sequence of events during differentiation and maturation may be obtained. This ex vivo-model is capable of studying the capacity of lymphocytes for differentiation and activation processes barely accessible in vivo. It may also be expected to represent an interesting tool for measuring the capacity for maturation and differentiation in the blood of children of different ages under normal and pathological conditions ex vivo. In addition, substance-induced effects may be studied in vitro with this approach on immature cells from newborn, or infants during culturing. Teratogenesis Carcinog. Mutagen. 20:171-193, 2000.