Ion transport across CF and normal murine olfactory and ciliated epithelium.

The nasal epithelium of the cystic fibrosis (CF) mouse has been used extensively in CF research because it exhibits ion transport defects similar to those of human CF airways. This tissue is composed of approximately 50% olfactory (OE) and approximately 50% ciliated epithelium (CE), and on the basis of previous observations, we hypothesized that a significant fraction of the bioelectric signals from murine nasal tissue may arise from OE rather than CE, while CE is the target tissue for CF gene therapy. We compared the bioelectric properties of isolated OE from the nasal cavity and CE from the nasopharynx in Ussing chamber studies. Hyperabsorption of Na(+) [amiloride response; CF vs. wild type (WT)] was approximately 7.5-fold greater in the OE compared with the CE. The forskolin response in native tissues did not reliably distinguish genotypes, likely due to a cyclic nucleotide-gated cation conductance in OE and a calcium-mediated Cl(-) conductance in CE. By potential difference assay, hyperabsorption of Na(+) (CF vs. WT) and the difference in response to apical 0 Cl(-) buffer (CF vs. WT) were approximately 2-fold greater in the nasal cavity compared with the nasopharynx. Our studies demonstrate that in the CF mouse, both the hyperabsorption of Na(+) and the Cl(-) transport defect are of larger magnitude in the OE than in the CE. Thus, while the murine CF nasal epithelium is a valuable model for CF studies, the bioelectrics are likely dominated by the signals from the OE, and assays of the nasopharynx may be more specific for studying the ciliated epithelium.

Expression of CFTR from a ciliated cell-specific promoter is ineffective at correcting nasal potential difference in CF mice.

Successful gene therapy will require that the therapeutic gene be expressed at a sufficient level in the correct cell type(s). To improve the specificity of gene transfer for cystic fibrosis (CF) and other airway diseases, we have begun to develop cell-type specific promoters to target the expression of transgenes to specific airway cell types. Using a FOXJ1 promoter construct previously shown to direct transgene expression specifically to ciliated cells, we have generated transgenic mice expressing human cystic fibrosis transmembrane conductance regulator (CFTR) in the murine tracheal and nasal epithelia. RNA analysis demonstrated levels of CFTR expression is greater than or equal to the level of endogenous mouse CFTR. Immunoprecipitation and western blotting demonstrated the production of human CFTR protein, and immunochemistry confirmed that CFTR was expressed in the apical region of ciliated cells. The transgenic animals were bred to CFTR null mice (Cftr(tm1Unc)) to determine if expression of CFTR from the FOXJ1 promoter is capable of correcting the airway defects in Cl(-) secretion and Na(+) absorption that accompany CF. Isolated trachea from neonatal CF mice expressing the FOXJ1/CFTR transgene demonstrated a correction of forskolin-stimulated Cl(-) secretion. However, expression of human CFTR in ciliated cells of the nasal epithelia failed to significantly change the nasal bioelectrics of the CF mice.

Culture of murine nasal epithelia: model for cystic fibrosis.

The ion transport defects reported for human cystic fibrosis (CF) airways are reproduced in nasal epithelia of the CF mouse. Although this tissue has been studied in vivo using the nasal potential difference technique and as a native tissue mounted in the Ussing chamber, little information is available on cultured murine nasal epithelia. We have developed a polarized cell culture model of primary murine nasal epithelia in which the CF tissue exhibits not only a defect in cAMP-mediated Cl- secretion but also the Na+ hyperabsorption and upregulation of the Ca2+-activated Cl- conductance observed in human airways. Both the wild-type and CF cultures were constituted predominantly of undifferentiated cuboidal columnar cells, with most cultures exhibiting a small number of ciliated cells. Although no goblet cells were observed, RT-PCR demonstrated the expression of Muc5ac RNA after approximately 22 days in culture. The CF tissue exhibited an adherent layer of mucus similar to the mucus plaques reported in the distal airways of human CF patients. Furthermore, we found that treatment of CF preparations with a Na+ channel blocker for 7 days prevented formation of mucus adherent to epithelial surfaces. The cultured murine nasal epithelial preparation should be an excellent model tissue for gene transfer studies and pharmacological studies of Na+ channel blockers and mucolytic agents as well as for further characterization of CF ion transport defects. Culture of nasal epithelia from DeltaF508 mice will be particularly useful in testing drugs that allow DeltaF508 CFTR to traffic to the membrane.

Aflatoxicosis in turkey poults is prevented by treatment of naturally contaminated corn with ozone generated by electrolysis.

Previous studies have demonstrated that a novel source of ozone gas (O3) maybe used to chemically degrade numerous mycotoxins, including aflatoxin (AF) B1. Subsequent in vitro analyses demonstrated detoxification of AFB1, suggesting a potential method of remediate AF-contaminated grain. The objective of this study was to evaluate the capability of electrochemically produced ozone to degrade AFB1 in naturally contaminated whole kernel corn and confirm detoxification in turkey poults. Corn was procured from the southern coastal areas of Texas and HPLC revealed 1,220 +/- 73.3 ppb AFB1. Control and contaminated corn were treated for 92 h with O3 at 200 mg/min in 30 kg batches; greater than 95% reduction of AFB1 in contaminated corn was achieved. One-day-old female turkey poults were fed 1) control corn, 2) control corn + O3, 3) AFB1 corn, or 4) AFB1 corn + O3 mixed in rations (46% by wt.) and consumed ad libitum for 3 wk. When compared with controls, turkeys fed AFB1 corn had reduced body weight gain and relative liver weight, whereas turkeys fed control corn + O3 or AFB1 corn + O3 did not differ from controls. Furthermore, alterations in the majority of relative organ weight, liver discoloration, serum enzyme activity, hematological parameters, and blood chemistry caused by AFB1 were eliminated (no difference from controls) by treatment with O3. These data demonstrate that treatment of contaminated corn with electrochemically produced O3 provided protection against AFB1 in young turkey poults. It is important to note that treatment of control corn with O3 did not alter the performance of the turkey poults.

Oxidative degradation and detoxification of mycotoxins using a novel source of ozone.

Practical methods to degrade mycotoxins using ozone gas (O3) have been limited due to low O3 production capabilities of conventional systems and their associated costs. Recent advances in electrochemistry (i.e. proton-exchange membrane and electrolysis technologies) have made available a novel and continuous source of O3 gas up to 20% by weight. It is possible that the rapid delivery of high concentrations of O3 will result in mycotoxin degradation in contaminated grains--with minimal destruction of nutrients. The major objectives of this study were to investigate the degradation and detoxification of common mycotoxins in the presence of high concentrations of O3. In this study, aqueous equimolar (32 microM) solutions of aflatoxins B1 (AfB1), B2 (AfB2), G1 (AfG1), G2 (AfG2), cyclopiazonic acid (CPA), fumonisin B1 (FB1), ochratoxin A (OA), patulin, secalonic acid D (SAD) and zearalenone (ZEN) were treated with 2, 10 and/or 20 weight% O3 over a period of 5.0 min and analysed by HPLC. Results indicated that AfB1 and AfG1 were rapidly degraded using 2% O3, while AfB2 and AfG2 were more resistant to oxidation and required higher levels of O3 (20%) for rapid degradation. In other studies, patulin, CPA, OA, SAD and ZEN were degraded at 15 sec, with no by-products detectable by HPLC. Additionally, the toxicity of these compounds (measured by a mycotoxin-sensitive bioassay) was significantly decreased following treatment with O3 for 15 sec. In another study, FB1 (following reaction with O3) was rapidly degraded at 15 sec, with the formation of new products. One of these appeared to be a 3-keto derivative of FB1. Importantly, degradation of FB1 did not correlate with detoxification, since FB1 solutions treated with O3 were still positive in two bioassay systems.

Parasuicide and the lunar cycle.

Admissions to hospital following parasuicide in one city over eighteen years exhibit a cyclical variation apparently synchronized with the lunar quarters. The effect would only account for approximately 0.7 out of the average of 46 parasuicides per 100,000 adults per lunar cycle (95% CI 0.6-0.8), and fails to reach statistical significance.

The P300 component of the auditory event-related potential in monozygotic and dizygotic twins.

The P300 component of the auditory event-related potential was recorded from 10 pairs of monozygotic (MZ) twins and 11 pairs of dizygotic (DZ) twins during an auditory discrimination task. The within-pair similarity of P300 latency was significantly greater in MZ than in DZ twins. The within-pair similarity of P300 amplitude was not significantly greater in MZ than in DZ twins. These results support previous findings suggesting that P300 latency is largely genetically determined.

A new photocatalytic material based on algal cells.

Metallic platinum was deposited at surfaces of intracellular photosynthetic membranes of whole cells of a cyanobacterium (blue-green alga). The deposited platinum particles acted as a catalyst for generation of hydrogen from photosynthetic decomposition of water in the absence of other exogenous electron transfer agents. This technique represents a means of placing metal catalysts in contact with intracellular structures of microorganisms.

The discrete dynamics of symmetric competition in the plane.

We consider the generalized Lotka-Volterra two-species system xn + 1 = xn exp(r1(1 - xn) - s1yn) yn + 1 = yn exp(r2(1 - yn) - s2xn) originally proposed by R. M. May as a model for competitive interaction. In the symmetric case that r1 = r2 and s1 = s2, a region of ultimate confinement is found and the dynamics therein are described in some detail. The bifurcations of periodic points of low period are studied, and a cascade of period-doubling bifurcations is indicated. Within the confinement region, a parameter region is determined for the stable Hopf bifurcation of a pair of symmetrically placed period-two points, which imposes a second component of oscillation near the stable cycles. It is suggested that the symmetric competitive model contains much of the dynamical complexity to be expected in any discrete two-dimensional competitive model.

Complete chaos in a simple epidemiological model.

A first-order difference equation which constitutes a simple model for a lethal parasite-host interaction is studied. Completing a study initiated by May and Anderson, the dynamics are shown to be completely chaotic.

Image analysis of atypical bronchial squamous epithelial cells. A quantitative and qualitative examination of squamous cell carcinogenesis.

Atypical bronchial squamous epithelial cells from cigarette smokers were digitized using a high-resolution microphotometric scanning system operating at a wavelength of 530 nm. All cells used for the purpose of developing a training set as well as those used as a test set were classified by a team of three cytopathologists. Reduction in the dimensionality of the feature space and cell classification were performed using a new system known as the atypia status index (ASI); this dual-purpose system will specifically classify cells into five distinct classes of progressive atypia as well as quantitate the degree of atypicality of each individual cell. The distribution of cellular ASI values was arranged from lowest to highest and plotted for each subject as a profile. The ASI range has well-defined intervals, representing the five different atypia classes used in the study (squamous metaplasia, mild atypia, moderate atypia, severe atypia and carcinoma). Subjects at different stages of carcinogenesis were found to have significantly different profiles (P less than 0.01). Changes in visual morphologic characteristics and their relation to changes in ASI values for the five different classes of atypia are presented.

A random walk model of cellular kinetics.

The sorting out of biological cell mixtures into clusters of one cell type is modelled as a consequence of random cell and cluster motion, during which cells of like type cohere upon collision. After a description of the model and its motility rules, the results of several computer simulation studies are analysed and compared with both laboratory data and certain theoretical predictions. The model is found to be more biologically realistic than previous models with similar results. Suggestions for further reserach are discussed. An Appendix contains details about the data structures and algorithms employed in the simulation.

Growth-rate periodicity of Streptomyces levoris during space flight.

Streptomyces levoris Kras was used is an experimental test micro-organism during the Apollo Soyuz Test Project to study alternating vegetative mycelial and spore ring periodicity during space flight. Four cultures were launched in each of the spacecrafts (Apollo and Soyuz). During the joint space-flight activities, two cultures from each spacecraft were exchanged. Selected duplicate cultures were maintained as controls in both the USA and the USSR. Spore ring morphology was periodically documented by photographing the specimens at approximately 12-hr intervals during the pre-, in-, and post-flight periods of the experiment. A decreased growth-rate periodicity in all but one of the eight space-flight cultures was in part attributed to the reduced temperature in the spacecraft. One of the eight cultures grew at a faster rate in the reduced temperature environment of Apollo than did the ground controls. Three of the space-flight cultures developed double spore rings during the immediate post-flight period. This anomaly was attributed to re-entry into the earth's gravity. The absence of spores in portions of one ring formed during space flight may have been caused by nutritional defects or media abnormality. Extensive studies will be required to elucidate the cause of this detect with certainty. There was no visible evidence of wedges in the cultures which would suggest naturally occurring or radiation-induced mutagenic alteration during space flight.