Basophilic “Dark Cells:” Optimal Conditions for Toluidine Blue Staining

Differential staining of basophilic “dark cells” of epithelia in thick epoxy sections with toluidine blue is critically affected by fixation and embedding conditions. Optimal conditions are described.     

The z-model — A proposal for spatial and temporal modeling of visual threshold perception

By considering only the modulation transfer functions of stationary, uniformly moved, and time modulated sinusoidal gratings it is possible to derive a simple model, the z-model, for the spatio-temporal frequency behaviour of one-dimensional patterns. The transmission function of this model is a band pass function of a single coordinate z, which is a quadratic form of the spatial and temporal frequencies (rotational symmetry with respect to space and time). The model is determined by only three constants. Optionally a time phase which accounts for delay and phase distortion can be added. This model can also be derived from reaction time measurements for switched on sinusoidal gratings. With this model the response of a wide variety of spatio-temporal patterns have been calculated and compared with measured threshold data. For two-dimensional patterns orientational filtering has to be added to the model leading to a further parameter. This model predicts satisfactorily the threshold modulation for a great variety of arbitrary spatio-temporal patterns. However the absolute threshold value for aperiodic transient patterns differs slightly in direction of smaller sensitivity as compared with periodic stationary patterns. This suggests that the peak detection scheme usually used in threshold detection modeling should be replaced by an integrative mechanism.     

ENZYME ACTIVITY AND CELL DIFFERENTIATION IN NEUROSPORA

Zalokar , Marko . (Yale U., New Haven, Conn.) Enzyme activity and cell differentiation in Neurospora. Amer. Jour. Bot, 46(7): 555–559. Illus. 1959.—Morphological differences were observed in vegetative cells of Neurospora of different ages and in different parts of the mycelium. The surface layer of mycelium grown in standing cultures could be separated from the deep layer. The first contained most of the growing hyphae rich in protoplasm, while the second contained heavily vacuolated hyphae laden with fat droplets. Specific activities of several enzymes were studied in conidia, young hyphae, and the surface and deep layers of mature mycelium. Succinic dehydrogenase was low in conidia and about 10 times more active in mature mycelium. The surface layer had twice the activity of the deep layer. Aldolase increased about 3 times after the germination of conidia; it was slightly lower in the surface than in the deep layer of mycelium. Tryptophan synthetase exhibited only small differences between conidia and mycelium and was slightly lower in the surface than in the deep layer. β-galactosidase was formed in appreciable amounts only after prolonged growth and had a much higher specific activity in the deep layer. The results were discussed in connection with cell differentiation and aging.     

GROWTH AND DIFFERENTIATION OF NEUROSPORA HYPHAE

Zalokar , Marko . (Yale U., New Haven, Conn.) Growth and differentiation of Neurospora hyphae. Amer. Jour. Bot. 46(8): 602–610. Illus. 1959.—Neurospora hyphae grow only at their tips. New protoplasm is formed throughout the hyphae and transported to the tips by active cytoplasmic streaming. This enables the fungus to grow at a prodigious rate. The use of radioactive precursors demonstrated that the rate of protein and ribonucleic acid formation does not decrease substantially in hyphal regions distant from the growing point. Apical growth of hyphae results in a morphological differentiation into young regions filled with cytoplasm and dense with nuclei and old regions, vacuolated and laden with fat droplets. Cytochemical methods were used to study the biochemical differentiation of hyphae and attention was focused on the growing region. It was found that hyphal tips, i.e., the first 50–100 μ of the growing end of the hypha, differed substantially from the rest of the young hyphae. Hyphal tips were slightly richer in proteins and significantly richer in protein-bound sulfhydryl groups and ribonucleic acid. Glycogen was absent at the tips. Peroxidase increased at the tips, while cytochrome oxidase and succinic dehydrogenase decreased. Alkaline phosphatase exhibited strong activity in a short region behind the extreme tip. β-galactosidase, in a few cases, was more active at the tips than in the rest of the hyphae. The results of cytological and cytochemical investigations are discussed in connection with the problem of cellular differentiation.     

Massively parallel sequencing of single cells by epicPCR links functional genes with phylogenetic markers

Many microbial communities are characterized by high genetic diversity. 16S ribosomal RNA sequencing can determine community members, and metagenomics can determine the functional diversity, but resolving the functional role of individual cells in high throughput remains an unsolved challenge. Here, we describe epicPCR (Emulsion, Paired Isolation and Concatenation PCR), a new technique that links functional genes and phylogenetic markers in uncultured single cells, providing a throughput of hundreds of thousands of cells with costs comparable to one genomic library preparation. We demonstrate the utility of our technique in a natural environment by profiling a sulfate-reducing community in a freshwater lake, revealing both known sulfate reducers and discovering new putative sulfate reducers. Our method is adaptable to any conserved genetic trait and translates genetic associations from diverse microbial samples into a sequencing library that answers targeted ecological questions. Potential applications include identifying functional community members, tracing horizontal gene transfer networks and mapping ecological interactions between microbial cells.     

Numerical simulation of iontophoresis in the drug delivery system

The architecture and composition of stratum corneum act as barriers and limit the diffusion of most drug molecules and ions. Much effort has been made to overcome this barrier and it can be seen that iontophoresis has shown a good effect. Iontophoresis represents the application of low electrical potential to increase the transport of drugs into and across the skin or tissue. Iontophoresis is a noninvasive drug delivery system, and therefore, it is a useful alternative to drug transportation by injection. In this study, we present a numerical model and effects of electrical potential on the drug diffusion in the buccal tissue and the stratum corneum. The initial numerical results are in good comparison with experimental observation. We demonstrate that the application of an applied voltage can greatly improve the efficacy of localized drug delivery as compared to diffusion alone.     

Isolation and characterization of cDNA clones corresponding with mRNAs that accumulate during auxin-induced lateral root formation

Lateral root formation in root cultures of Arabidopsis thaliana can be initiated by exogenous addition of auxin. In order to find cDNA clones of which the corresponding mRNAs accumulate during this process, a cDNA library was constructed from root cultures treated with the active auxin 1-naphthaleneacetic acid (1- NAA). Differential screening of this library with cDNA probes derived from mRNA populations isolated from root cultures treated with 1-NAA and the inactive analogue 2-naphthaleneacetic acid (2-NAA) led to the isolation of four cDNA clones, designated AIR1, AIR3, AIR9 and AIR12. Accumulation of the mRNAs starts between 4 and 8 h and continues till at least 24 h after addition of an active auxin. Sequence analysis revealed that AIR1 encodes a protein that is related to a large family of proteins that consist of a proline-rich or glycine- rich N-terminus and a hydrophobic, possibly membrane spanning C- terminus. The putative function of these proteins is coupling of the cell wall to the plasma membrane. Surprisingly, AIR1 lacks the proline-rich or glycine-rich N-terminus which is thought to be important for interaction with the cell wall. AIR3 encodes a subtilisin-like serine protease which is believed to be active outside the plant cell. Although AIR9 and AIR12 do not show any significant homology to sequences in the database, they are also predicted to function outside the cell. Our screening thus indicates that a variety of genes encoding extracellular proteins are activated during auxin-induced lateral root formation.     

Effect of a permanent magnetic field on wing size parameters in Drosophila melanogaster.

The width and length variability of both D. melanogaster wings were measured in samples of flies after two and six generations under a permanent magnetic field of 35 mT. While in earlier generations under exposure sexes differ in the size response, later they both show a decrease in the wing size under the magnetic field exposure if compared to the control. The bilateral asymmetry in wings as a potential indicator of developmental instability does not increase significantly under exposure. The ecological and adaptive implications of the change of the magnetic intensity as an environmental factor generating stress in populations is discussed.