Three-dimensional perfused cell culture

Compelling evidence suggests the limitation and shortcomings of the current and well established cell culture method using multi-well plates, flasks and Petri dishes. These are particularly important when cell functions are sensitive to the local microenvironment, cell–cell and cell–extracellular matrix interactions. There is a clear need for advanced cell culture systems which mimic in vivo and more physiological conditions. This review summarises and analyses recent progress in three dimensional (3D) cell culture with perfusion as the next generation cell culture tools, while excluding engineered tissue culture where three dimensional scaffold has to be used for structural support and perfusion for overcoming mass transfer control. Apart from research activities in academic community, product development in industry is also included in this review.     

The effects of maturation on self-induced dynamic body sway frequencies of girls performing acrobatics or classical dance

We investigated the effects of maturation on the dynamic body sways of healthy girls. Prepubertal and postpubertal girls practising professional physical activities requiring a good ability to maintain equilibrium (acrobats and dancers) were asked to stand on a free seesaw platform and the results compared to those for untrained age-matched girls. This platform (stabilometer) allows self-induced body sways. Stabilograms were obtained by a double integration of the angular acceleration from the recordings of the platform sways made with an accelerometer. Fast Fourier transform processing of stabilograms allowed spectral frequency analysis. The total spectrum energy and the energies of three frequency bands (0–0.5 Hz, 0.5–2 Hz, 2–20 Hz) were determined. ANOVA showed that, for all groups of different equilibrium activity and independent of visual input, prepubertal girls had higher energy values than postpubertal girls in the 0- to 0.5-Hz band whereas the opposite was true for 0.5- to 2-Hz band. Ballet dancers were more dependent than acrobats on visual inputs for the regulation of their postural control but were less dependent than untrained girls at both ages. Maturation seemed to shift body sways towards higher frequencies and the utilization of the cues of postural control was different according to the type of equilibrium activity practised by the subjects. Accepted: 7 February 1997     

Volatile emission of different plant parts and fruit development from Italian cherry plums (Prunus cerasifera and P. cerasifera ‘Pissardii’)

The chemical composition of volatiles emitted in vivo from different plant parts of P. cerasifera and P. cerasifera ‘Pissardii’ were collected during the entire biological cycle of the plant growth and analyzed by gas chromatography mass spectrometry (GC-MS) after solid phase micro-extraction (SPME). All the data were submitted to multivariate statistical analysis evidencing many differences amongst the selected plant parts and growth stages. A total of 136 compounds were identified corresponding to 90.1–99.6% of the whole aroma profile of cherry plum samples. Non-terpenes were the most abundant class of constituents present in the volatile emission of all analyzed samples. In particular, the aroma of both fruit stages was mainly characterized by alcohols and esters in different amounts.     

Electrophoretic profiles of cyanobacterial membrane polypeptides showing heme-dependent peroxidase activity

The photosynthetic membranes of Anacystis nidulans R2 were examined electrophoretically following solubilization with lithium dodecyl sulfate. Electrophoresis yielded six prominent chlorophyll-containing bands. In addition, five polypeptides were observed which possessed heme-dependent peroxidase activity, monitored by incubating gels with 3,3′,5,5′-tetramethylbenzidine plus hydrogen peroxide. One such polypeptide, at 105 kdaltons, was removed by repeated washing of the membranes. Four remaining peroxidase-active polypeptides were observed at 7.2, 13.5, 18.5 and 33 kdaltons. Further examination of these four polypeptides yielded the following results. (1) The mobility of the 33 kdalton polypeptide was altered from 29 to 33 kdaltons upon heating (70°C) during membrane solubilization. (2) All four polypeptides showed stable heme-protein associations in the presence of 8 M urea; however, in the presence of urea, alterations in protein mobility were observed for each poly-peptide and only two (at 13.5 and 33 kdaltons) showed peroxidase activity following heating (70°C) during membrane solubilization. (3) The presence of thiols during membrane solubilization at 0°C was required to observe peroxidase activity at 7.2 kdaltons. These results, when compared to known properties of isolated cytochromes, suggest that the four polypeptides characterized here correspond to the subunits of photosynthetic cytochromes. Electrophoretic assessment of maize mutants lacking cytochrome f and b₆ activity supports this suggestion.     

Physiological integration in Cassia fasciculata Michx.: inflorescence removal and defoliation experiments

Summary In the annual herb Cassia fasciculata virtually every leaf subtends an axillary inflorescence. We examined the degree to which these leaf-inflorescence units (reproductive nodes) were physiologically independent of each other in the production of flowers, fruits, and seeds. Removal of up to 4 of every 5 inflorescences resulted in substantial increases in fruit and seed production by remaining, intact reproductive nodes. These increases nearly compensated for and manipulated reproductive nodes were associated with different vascular strands. When 2 of every 3 leaves were removed, fruit and seed production were reduced at both intact and defoliated reproductive nodes. Taken together, these results suggest that neighboring reproductive nodes in C. fasciculata are not physiologically independent of one another, and that competition among fruits and seeds for parental resources occurs over several reproductive nodes.Scientific contribution no. 1595 from the New Hampshire Agricultural Experiment Station     

Clone bank and physical and genetic map of potato chloroplast DNA

Summary Clone banks of PvuII, BamHI and XhoI fragments were generated of the Solanum tuberosum cv Katahdin plastome. These clone banks, in conjunction with molecular hybridization to tobacco ctDNA probes, were used to construct a physical map of potato ctDNA. The potato plastome was found to be a circular molecule of 155–156 Kbp containing two inverted repeat regions of 23–27 Kbp. The arrangement of restriction sites is very similar to that of other Solanaceae plastomes. Heterologous hybridization to known ctDNA encoded gene probes from tobacco allowed us to establish a genetic map of the potato chloroplast genome. The arrangement of these genes on the potato plastome resembles that on most higher plant ctDNAs.     

酸雨对翅荚木的生理反应及其机理的研究

本文指出了酸雨对翅荚木细胞透性、pH值、等电点、耐酸力均有显著的影响。其影响是随酸雨pH值的下降和浸没时间的延长而增加。发芽种子的抗坏血酸氧化酶、多酚氧化酶的活性,呼吸强度和可溶性糖、粗脂肪和蛋白质的含量,也是随酸雨pH值的下降而降低。叶绿素总量、叶绿素a,b和a/b的比值,pH 5处理与对照无差异,低于pH4以下的处理叶绿素a含量则大幅度减少。叶绿素b含量则较稳定。根系的活力也是随酸雨pH值下降而急剧减少。酸雨对翅荚木种子的发芽率、出苗率及胚和幼苗的生长,都表现有不同程度的抑制效应,酸雨对叶片的伤害,也是随酸雨pH值的降低、喷洒次数的增多和浸没时间的延长而加重。     

Scaling physiological measurements for individuals of different body size

This paper examines how selected physiological performance variables, such as maximal oxygen uptake, strength and power, might best be scaled for subject differences in body size. The apparent dilemma between using either ratio standards or a linear adjustment method to scale was investigated by considering how maximal oxygen uptake (l.min-1), peak and mean power output (W) might best be adjusted for differences in body mass (kg). A curvilinear power function model was shown to be theoretically, physiologically and empirically superior to the linear models. Based on the fitted power functions, the best method of scaling maximum oxygen uptake, peak and mean power output, required these variables to be divided by body mass, recorded in the units kg 2/3. Hence, the power function ratio standards (ml.kg-2/3.min-1) and (W.kg-2/3) were best able to describe a wide range of subjects in terms of their physiological capacity, i.e. their ability to utilise oxygen or record power maximally, independent of body size. The simple ratio standards (ml.kg-1.min-1) and (W.kg-1) were found to best describe the same subjects according to their performance capacities or ability to run which are highly dependent on body size. The appropriate model to explain the experimental design effects on such ratio standards was shown to be log-normal rather than normal. Simply by taking logarithms of the power function ratio standard, identical solutions for the design effects are obtained using either ANOVA or, by taking the unscaled physiological variable as the dependent variable and the body size variable as the covariate, ANCOVA methods.