Modeling of contact-inhibited animal cell growth on flat surfaces and spheres

A model of contact-inhibited growth of cells on flat and spherical surfaces is presented. It shows that contact inhibition does not significantly affect the calculated growth rate of cells unless they are allowed to multiply a large amount from the original seeding density. Microcarriers seeded at low densities require long times to reach confluence because contact inhibition becomes important. In systems with both growth and separate cell death, the equilibrium fraction of holes in the confluent monolayer is below 8% if the death rate is less than half the growth rate, but increases rapidly as the death rate increases from that value.     

Transient shear stresses on a suspension cell in turbulence

A model flow field representative of Kolmogorov eddies in turbulence is proposed, and its two parameters are expressed in terms of the known bioreactor variables epsilon, the rate of turbulent power dissipation, and nu, the fluid kinematic viscosity. The trajectory through this flow field of a small sphere representing a cell is determined, and from that the time-varying local shear rate can be found. This allows calculation of the shear stress at any point on the sphere's surface as it rotates in and moves through the model eddy. The maximum shear stress imposed on the cell by the surrounding turbulence has a range of 0.5-5 dyn/cm(2), and can be estimated by 5.33rho(epsilonnu)(1/2). The shear stress has two major frequency components with ranges of 1-4 and 20-80 s(-1); the higher frequency component is estimated by 0.678(epsilon/nu)(1/2). The rotation of the direction of the shear stress vector at each point on the cell's surface is also discussed. Two ways in which external stresses might affect cell growth are proposed.     

Regional differences in capillary density of the normal human dermis

Quantitative study of the capillary density of normal dermis of 113 biopsy specimens taken from 20 body sites from 6 cadavers was carried out. Histologic quantification of dermal capillaries using the alkaline phosphatase method to delineate vascular endothelium and automated image analysis demonstrates statistically significantly greater capillary density in the head-face/neck region both in the papillary and reticular dermis than in the lower parts of the body. The smallest numbers of the dermal capillary density were noted in the calf and shin areas. These data support the hypothesis that random-pattern skin flaps can be safely raised longer in the face and neck than in other parts of the body.     

Evidence of structural and functional adaptation in epilithon exposed to zinc

Artificial streams were used to study the effect of chronic zinc dosing (0.00, 0.05 or 1.00 mg Zn l^–1) on epilithic communities during summer and early fall, 1984 and 1985. Following zinc addition, epilithon was periodically measured for ¹⁴C-glucose and ¹⁴C-glutamate respiration, ash-free dry weight (AFDW), protein, carbohydrate, chlorophyll a (Chl a), pheophytin a (pheo a), and colony forming units (CFU) on zinc amended and unamended media. Transient peaks in glucose respiration rates mg^–1 protein and AFDW occurred within 5 to 10 days in streams dosed with 1.0 mg Zn l^–1 and coincided with decreases in AFDW, protein, carbohydrate, Chl a, and pheo a. Respiration of glutamate did not show a significant response to dosing ( = 0.05). Epilithic plate counts demonstrated more total and zinc-tolerant CFU in dosed than in control epilithon after 20 days. After 30 days, greater biomass and lower protein: carbohydrate ratios were evident in epilithon dosed with 1.0 mg Zn l^–1 compared to unamended treatments. Peaks in glucose respiration suggested the presence of zinc-tolerant heterotrophs transiently capable of increased respiration of glucose but not glutamate. Concurrently, minimal but measurable biomass inferred the persistence of zinc-tolerant microorganisms in zinc-dosed epilithon. By the end of the studies, results indicated that metal-tolerant populations replaced metal-sensitive organisms under conditions of elevated zinc concentration. These zinc adapted communities had lower potential value as a food source to grazers (low protein: carbohydrate ratio) and were no longer distinctive with respect to glucose respiration.     

Colonization,variability, and the use of substratum-filled trays for biomonitoring benthic communities

Sampling variability and colonization rate of introduced substrates (plastic trays filled with pebble and cobble) in two southwestern Virginia streams are described. Substrates were rapidly colonized by aquatic macroinvertebrates, but colonization rates differed between years, possibly due to annual variability in macroinvertebrate abundance. To examine the applicability of using these substrates for biomonitoring benthic communities, trays were placed at several locations in a river receiving power plant discharges. Only six samples were necessary to detect a 15%reduction in macroinvertebrate density and a 12% reduction in number of taxa at effluent sites. Benthic communities established on rock-filled trays and multiplate samplers collected from the same stations during the same period were compared. Although multiplate samplers were more variable than rock trays and were selective for different taxa, both substrate types showed significant differences in community parameters among locations. Rock trays at all sites were dominated by Cheumatopsyche sp., whereas chironomids were more abundant on multiplate samplers. The relative abundance of mayflies was reduced at the effluent site on both substrate types.     

Mutational analysis of conserved nucleotides in a self-splicing group I intron.

We have constructed all single base substitutions in almost all of the highly conserved residues of the Tetrahymena self-splicing intron. Mutation of highly conserved residues almost invariably leads to loss of enzymatic activity. In many cases, activity could be regained by making additional mutations that restored predicted base-pairings; these second site suppressors in general confirm the secondary structure derived from phylogenetic data. At several positions, our suppression data can be most readily explained by assuming non-Watson-Crick base-pairings. In addition to the requirements imposed by the secondary structure, the sequence of the intron is constrained by "negative interactions", the exclusion of particular nucleotide sequences that would form undesirable secondary structures. A comparison of genetic and phylogenetic data suggests sites that may be involved in tertiary structural interactions.     

Ontogeny of the sexually dimorphic male nucleus in the preoptic/anterior hypothalamus of ferrets and its manipulation by gonadal steroids

A sexually dimorphic nucleus exists in the dorsal region of the ferret preoptic/anterior hypothalamic area (POA/AH), and is called the male nucleus of the POA/AH (MN-POA/AH) because it is found only in males. Development of the MN-POA/AH was studied in male ferrets, and for comparison a sexually nondimorphic ventral POA/AH nucleus was studied in both sexes. The MN-POA/AH was conspicuous in males as early as embryonic day 37 (E37) of a 41-day gestation, and its volume increased until postnatal day 56 (P56). No nucleus was present in the dorsal POA/AH of females at any age. The densities and average somal areas of cells in the dorsal POA/AH were similar in males and females at E33, before the MN-POA/AH could be visualized. However, at E37 and E41 dorsal cells were greater in density and/or somal area in males than in females, accounting for the appearance of a nucleus in males at these ages. To insure that the dorsal POA/AH nucleus seen in males at E37 and E41 was the presumptive MN-POA/AH present in adult males, pregnant ferrets were given progesterone and either implanted subcutaneously (s.c.) with testosterone (T) or ovariectomized and implanted s.c. with the aromatase inhibitor, 1,4,6-androstatriene-3,17-dione (ATD), on day 30 of gestation. As predicted from previous studies in which subjects were sacrificed in adulthood, formation of a dorsal POA/AH nucleus was promoted in female ferrets by T, and blocked in males by maternal ovariectomy and ATD treatment for animals sacrificed at E41. Much evidence suggests that behavioral sexual differentiation is accomplished in the male ferret between age E28 and P20. The MN-POA/AH is present and potentially functional in males during a considerable portion of this perinatal period.     

Revealing pancrustacean relationships: Phylogenetic analysis of ribosomal protein genes places Collembola (springtails) in a monophyletic Hexapoda and reinforces the discrepancy between mitochondrial and nuclear DNA markers

Background  

In recent years, several new hypotheses on phylogenetic relations among arthropods have been proposed on the basis of DNA sequences. One of the challenged hypotheses is the monophyly of hexapods. This discussion originated from analyses based on mitochondrial DNA datasets that, due to an unusual positioning of Collembola, suggested that the hexapod body plan evolved at least twice. Here, we re-evaluate the position of Collembola using ribosomal protein gene sequences.     

Genomic RNAi screening in Drosophila S2 cells: what have we learned about host-pathogen interactions?

The détente between pathogen and host has been of keen interest to researchers in spite of being exceedingly difficult to probe. Recently, new RNA interference (RNAi) technologies, in particular in Drosophila tissue culture cells, have made it possible to interrogate the genetics of host organisms rapidly, with nearly complete genomic coverage and high fidelity. Therefore, it is not surprising that the applications of RNAi to the study of host-pathogen interactions were among the first to be published and have already revealed many new insights into the hosts' role in infection. This review will highlight the application of RNAi screening to pathogen-host interactions in Drosophila cells and will reveal some of the lessons learned from this approach.     

Burn trauma alters calcium transporter protein expression in the heart.

We have shown previously that burn trauma produces significant cardiac dysfunction, which is first evident 8 h postburn and is maximal 24 h postburn. Because calcium handling by the cardiomyocyte is essential for cardiac function, one mechanism by which burn injury may cause cardiac abnormalities is via calcium dyshomeostasis. We hypothesized that major burn injury alters cardiomyocyte calcium handling through changes in calcium transporter expression. Sprague-Dawley rats were given either burn injury or no burn injury (controls). Cardiomyocyte intracellular calcium and sodium were quantified at various times postburn by fura 2-AM or sodium-binding benzofuran isophthalate fluorescent indicators, respectively. In addition, hearts freeze-clamped at various times postburn (2, 4, 8, and 24 h) were used for Western blot analysis using antibodies against the sarcoplasmic reticulum calcium-ATPase (SERCA), the L-type calcium-channel, the ryanodine receptor, the sodium/calcium exchanger, or the sodium-potassium-ATPase. Intracellular calcium levels were elevated significantly 8-24 h postburn, and intracellular sodium was increased significantly 4 through 24 h postburn. Expression of SERCA was significantly reduced 1-8 h postburn, whereas L-type calcium-channel expression was diminished 1 and 2 h postburn (P < 0.05) but returned toward control levels 4 h postburn. Ryanodine receptor protein was significantly reduced at 1 and 2 h postburn, returning to baseline by 4 h postburn. Sodium/calcium exchanger expression was significantly elevated 2 h postburn but was significantly reduced 24 h postburn. An increase in sodium-potassium-ATPase expression occurred 2-24 h postburn. These data confirm that burn trauma alters calcium transporter expression, likely contributing to cardiomyocyte calcium loading and cardiac contractile dysfunction.