Epithelium Expressing the E7 Oncoprotein of HPV16 Attracts Immune-Modulatory Dendritic Cells to the Skin and Suppresses Their Antigen-Processing Capacity

Antigen presenting cells (APCs) in skin can promote either antigen-specific effector functions or antigen tolerance, and thus determine clearance or persistence of cutaneous viral infections. Human papillomavirus (HPV) infections can persist in squamous epithelium in immunocompetent individuals, and some persisting HPV infections, particularly with HPV16, promote malignant epithelial transformation. Here, we investigate whether local expression of the HPV16 protein most associated with malignant transformation, HPV16-E7, affects the phenotype and function of APC subsets in the skin. We demonstrate an expanded population of Langerhans cells in HPV16-E7 transgenic skin with distinct cell surface markers which express immune-modulatory enzymes and cytokines not expressed by cells from non transgenic skin. Furthermore, HPV16-E7 transgene expression in keratinocytes attracts new APC subsets to the epidermis. In vivo migration and transport of antigen to the draining lymph node by these APCs is markedly enhanced in HPV16-E7 expressing skin, whereas antigen-processing, as measured by proteolytic cleavage of DQ-OVA and activation of T cells in vivo by APCs, is significantly impaired. These data suggest that local expression of HPV16-E7 in keratinocytes can contribute to persisting infection with this oncogenic virus, by altering the phenotype and function of local APCs.     

Characteristics of newt breeding sites

Breeding site characteristics have been studied for the three species of newt that occur in Britain, the Palmate ( Triturus helveticus (Razoumowski)), Smooth ( T. vulgaris (L.)) and Warty ( T. cristatus (Laurenti)). The Warty newt was seldom found in the absence of the much commoner Smooth newt, but seemed to prefer sites that were relatively large and deep and that had a high proportion of open water surface. All three species tended to breed in ponds having abundant aquatic vegetation. Smooth newts, unlike Palmate newts, were rarely encountered in water with pH <6. The Smooth newt tended to be found in water with relatively high concentrations of metals, while the reverse was true for the Palmate newt. Over Britain, Smooth and Warty newts are relatively less common in soft water areas, while the Palmate is less common in hard water areas. Possible reasons for these associations are discussed.     

Helical motion analysis of the knee--I. Methodology for studying kinematics during locomotion

A technique for investigating the three-dimensional kinematics of knee motion during dynamic functional tasks has been developed. It involves the combined usage of a six degree of freedom goniometer and helical motion analysis. A detailed procedure for coordinate system alignment and calibration must be followed. Once established this entire procedure is routinely implementable. Ensemble averages from multiple walking strides reveal that this technique is sensitive enough to differentiate between the kinematics of an uninjured and injured knee.     

Effect of serial intrauterine dimethyl sulfoxide infusions on the incidence of periglandular fibrosis in category II horse endometria

High concentrations of dimethyl sulfoxide (DMSO) have been shown to cause gross structural changes in collagen fibers in vitro. As endometrial fibrosis is a major cause of infertility in mares, the effect of serial DMSO infusions on periglandular fibrosis of Category II endometria was investigated. Initially, six mares with Category I endometria were serially infused with DMSO to determine whether such therapy would incite fibrosis. Four mares, serially infused with saline, served as controls in each experiment. One hundred milliliters of 75% DMSO was infused initially, followed by the same volume of 25% DMSO daily for 6 d. Endometrial biopsies obtained 21 d after the last infusion displayed no significant difference in the incidence of periglandular fibrosis when compared with the Day 0 biopsies. Examination of treated uteri at necropsy showed no gross evidence of adhesions or ulcerations.     

Differential survival of chaparral seedlings during the first summer drought after wildfire

Summary Big Pod Ceanothus (Ceanothus megacarpus) is an obligate seeder after fire; Laurel Sumac (Rhus laurina) is primarily a resprouter after fire. Both species commonly occur together in mixed stands and are dominant members of the coastal chaparral of southern California. We compared the mean survival of post-fire seedlings of each species during the first summer drought after fire and found C. megacarpus to have a mean survival of 54% while R. laurina had a mean survival of only 0.1%. Rooting dephs were similar between species but predawn water potentials and leaf temperatures were higher for R. laurina seedlings. Leaf temperatures for R. laurina reached a mean value of 46.8° C on hot, summer days, about 5° C higher than seedlings of C. megacarpus. By the end of the first growing season, 92% of all C. megacarpus seedlings had suffered herbivory compared to only 17% of all R. laurina seedlings. Herbivory did not appear to be the immediate cause of seedling mortality. Transect data indicated that full recovery of prefire species composition and density at our study site was likely but the mode of recovery was different for the species examined. R. laurina recovered primarily by sprouting, C. megacarpus totally by seedling establishment and a third species, Adenostoma fasciculatum (chamise), by a combination of sprouting and seedling establishment. We attribute the higher mortality of R. laurina seedlings to the greater sensitivity of its tissue to water stress. It may be that differential survival of shrub seedlings and differential modes of reestablishment after fire play an important role in maintaining species diversity in the chaparral communities of coastal, southern California.     

Kinetics and mechanics of stem gravitropism in Coprinus cinereus

Using video recordings we have completed the first kinetic analysis of mushroom stem gravitropism. The stem became gravireceptive after completion of meiosis, beginning to bend within 30 minutes of being placed horizontal. Stem bending first occurred in the apical 15% of its length, then the position of the bend moved rapidly towards the base, traversing 40% of stem length in 2.5 h. Meanwhile, the stem elongated by 25%, mostly in its upper half but also in basal regions. If the apex was pinned horizontally the stem base was elevated but overshot the vertical, often curling through more than 300 degrees. When the base was pinned to the horizontal (considered analogous to the normal situation), 90% of the initial bend was compensated as the stem brought its apex accurately upright, rarely overshooting the vertical. The apex had to be free to move for this curvature compensation to occur. Stems transferred to a clinostat after some minutes gravistimulation showed curvature which increased with the length of initial gravistimulation, indicating that continued exposure to the unilateral gravity vector was necessary for continued bending. Such gravistimulated stems which bent on the clinostat subsequently relaxed back towards their original orientation. Reaction kinetics were unaffected by submergence in water, suggesting that mechanical events do not contribute, but submerged stems bent first at the base rather than apex. In air, the gravitropic bend appeared first near the apex and then moved towards the base, suggesting basipetal movement of a signal. In water, the pattern of initial bending was changed (from apex to base) without effect on kinetics. Taken together these results suggest that bending is induced by a diffusing chemical growth factor (whose extracellular propagation is enhanced under water) which emanates from the apical zone of the stem. The apex is also responsible for regulating compensation of the bend so as to bring the tip to the vertical. The nature of this latter stimulus is unknown but it is polarized (the apex must be free to move for the compensation to occur) and it may not require reference to the unilateral gravity vector.     

Anaerobic C1 Metabolism of the O-Methyl-14C-Labeled Substituent of Vanillate

The O-methyl substituents of aromatic compounds constitute a C₁ growth substrate for a number of taxonomically diverse anaerobic acetogens. In this study, strain TH-001, an O-demethylating obligate anaerobe, was chosen to represent this physiological group, and the carbon flow when cells were grown on O-methyl substituents as a C₁ substrate was determined by ¹⁴C radiotracer techniques. O-[methyl-¹⁴C]vanillate (4-hydroxy-3-methoxy-benzoate) was used as the labeled C₁ substrate. The data showed that for every O-methyl carbon converted to [¹⁴C]acetate, two were oxidized to ¹⁴CO₂. Quantitation of the carbon recovered in the two products, acetate and CO₂, indicated that acetate was formed in part by the fixation of unlabeled CO₂. The specific activity of ¹⁴C in acetate was 70% of that in the O-methyl substrate, suggesting that only one carbon of acetate was derived from the O-methyl group. Thus, it is postulated that the carboxyl carbon of the product acetate is derived from CO₂ and the methyl carbon is derived from the O-methyl substituent of vanillate. The metabolism of O-[methyl-¹⁴C]vanillate by strain TH-001 can be described as follows: 3¹⁴CH₃OC₇H₅O₃ + CO₂ + 4H₂O → ¹⁴CH₃COOH + 2¹⁴CO₂ + 10H⁺ + 10e^- + 3HOC₇H₅O₃.     

Dependence of magnetic resonance image (MRI) intensity values on relaxation times, pulse intervals and other signal attenuation factors

Magnetic resonance image (MRI) pixel intensities were investigated using a phantom containing several uniform size chambers filled with solutions of known relaxation times, as well as head scans of patients and volunteers. Intensities were measured with a variety of pulse intervals typically used for imaging with spin echo, (SE) and inversion recovery (IR) sequences at 0.15 Tesla using the back projection (R-THETA) method, and at 0.27 Tesla using the 2-dimensional Fourier transform (2DFT) technique. The results were compared with the calculated dependence of MRI signal intensity on relaxation times and pulse interval parameters using the well known functions containing exponential forms. The experimental and the calculated pixel intensity time dependence did not always agree. We infer that factors other than the conventional functions for T1 and T2 signal decay are important. These factors may include the attenuation of the radiofrequency (RF) signals through inhomogenious lossy dielectric materials (e.g., tissues and organs), the location (coordinate) of the portion of the sample to be imaged relative to the RF coils, and the timing and amplitude of gradient pulses relative to the RF input and the detected signals. The flow velocity and diffusions are also important determinants of the signal from blood vessels and body fluids. We point out the necessity for further investigation toward more comprehensive understanding of MRI intensities.     

Analysis of chromosome 22 deletions in neurofibromatosis type 2-related tumors.

The neurofibromatosis type 2 (NF2) gene has been hypothesized to be a recessive tumor suppressor, with mutations at the same locus on chromosome 22 that lead to NF2 also leading to sporadic tumors of the types seen in NF2. Flanking markers for this gene have previously been defined as D22S1 centromeric and D22S28 telomeric. Identification of subregions of this interval that are consistently rearranged in the NF2-related tumors would aid in better defining the disease locus. To this end, we have compared tumor and constitutional DNAs, isolated from 39 unrelated patients with sporadic and NF2-associated acoustic neuromas, meningiomas, schwannomas, and ependymomas, at eight polymorphic loci on chromosome 22. Two of the tumors studied revealed loss-of-heterozygosity patterns, which is consistent with the presence of chromosome 22 terminal deletions. By using additional polymorphic markers, the terminal deletion breakpoint found in one of the tumors, an acoustic neuroma from an NF2 patient, was mapped within the previously defined NF2 region. The breakpoint occurred between the haplotyped markers D22S41/D22S46 and D22S56. This finding redefines the proximal flanking marker and localizes the NF2 gene between markers D22S41/D22S46 and D22S28. In addition, we identified a sporadic acoustic neuroma that reveals a loss-of-heterozygosity pattern consistent with mitotic recombination or deletion and reduplication, which are mechanisms not previously seen in studies of these tumors. This finding, while inconsistent with models of tumorigenesis that invoke single deletions and their gene-dosage effects, lends further support to the recessive tumor-suppressor model.