Regulation of the alpha-glucuronidase-encoding gene ( aguA) from Aspergillus niger.

The alpha-glucuronidase gene aguA from Aspergillus niger was cloned and characterised. Analysis of the promoter region of aguA revealed the presence of four putative binding sites for the major carbon catabolite repressor protein CREA and one putative binding site for the transcriptional activator XLNR. In addition, a sequence motif was detected which differed only in the last nucleotide from the XLNR consensus site. A construct in which part of the aguA coding region was deleted still resulted in production of a stable mRNA upon transformation of A. niger. The putative XLNR binding sites and two of the putative CREA binding sites were mutated individually in this construct and the effects on expression were examined in A. niger transformants. Northern analysis of the transformants revealed that the consensus XLNR site is not actually functional in the aguA promoter, whereas the sequence that diverges from the consensus at a single position is functional. This indicates that XLNR is also able to bind to the sequence GGCTAG, and the XLNR binding site consensus should therefore be changed to GGCTAR. Both CREA sites are functional, indicating that CREA has a strong influence on aguA expression. A detailed expression analysis of aguA in four genetic backgrounds revealed a second regulatory system involved in activation of aguA gene expression. This system responds to the presence of glucuronic and galacturonic acids, and is not dependent on XLNR.

Hyperactivity caused by a nitric oxide synthase inhibitor is countered by ultra-wideband pulses.

Potential action of ultra-wideband (UWB) electromagnetic field pulses on effects of N(G)-nitro- L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase (NOS), on nociception and locomotor activity was investigated in CF-1 mice. Animals were injected IP with saline or 50 mg/kg L-NAME and exposed for 30 min to no pulses (sham exposure) or UWB pulses with electric field parameters of 102+/-1 kV/m peak amplitude, 0.90+/-0.05 ns duration, and 160+/-5 ps rise time (mean+/-S.D.) at 600/s. Animals were tested for thermal nociceptive responses on a 50 degrees C surface and for spontaneous locomotor activity for 5 min. L-NAME by itself increased mean first-response (paw lift, shake, or lick; jump) and back-paw-lick response latencies and mean locomotor activity. Exposure to UWB pulses reduced the L-NAME-induced increase in back-paw-lick latency by 22%, but this change was not statistically significant. The L-NAME-induced hyperactivity was not present after UWB exposure. Reduction and cancellation of effects of L-NAME suggest activation of opposing mechanism(s) by the UWB pulses, possibly including increase of nitric oxide production by NOS. The action, or actions, of UWB pulses appears to be more effective on locomotor activity than on thermal nociception in CF-1 mice.

Shwachman syndrome: phenotypic manifestations of sibling sets and isolated cases in a large patient cohort are similar.

OBJECTIVES: With the use of clinical data from a large international cohort, we evaluated and compared affected siblings and isolated cases. STUDY DESIGN: Data from 116 families were collected, and patients conforming to our predetermined diagnostic criteria were analyzed. Phenotypic manifestations of affected siblings and singletons were compared with the use of t tests, Wilcoxon scores, and chi2 analysis. RESULTS: Eighty-eight patients (33 female, 55 male; median age 5.20 years) fulfilled our predetermined diagnostic criteria for Shwachman syndrome; 63 patients were isolated cases, and 25 affected siblings were from 12 multiplex families. Steatorrhea was present in 86% (57 of 66), and 91% (78 of 86) displayed a low serum trypsinogen concentration. Patients older than 4 years more often had pancreatic sufficiency. Neutropenia occurred in 98%, anemia in 42%, and thrombocytopenia in 34%. Myelodysplasia or cytogenetic abnormalities were reported in 7 patients. Short stature with normal nutritional status was a prominent feature. CONCLUSIONS: Clinical features among patients with Shwachman syndrome varied between patients and with age. Similarities in phenotype between isolated cases and affected sibling sets support the hypothesis that Shwachman syndrome is a single disease entity.

Frequency of micronuclei in the blood and bone marrow cells of mice exposed to ultra-wideband electromagnetic radiation.

PURPOSE: To investigate the extent of genetic damage in the peripheral blood and bone marrow cells of mice exposed to ultra-wideband electromagnetic radiation (UWBR). MATERIALS AND METHODS: CF-1 male mice were exposed to UWBR for 15 min at an estimated whole-body average specific absorption rate of 37 mW x kg(-1). Groups of untreated control and positive control mice injected with mitomycin C were also included in the study. After various treatments, half of the mice were killed at 18 h, and the other half at 24 h. Peripheral blood and bone marrow smears were examined to determine the extent of genotoxicity, as assessed by the presence of micronuclei (MN) in polychromatic erythrocytes (PCE). RESULTS: The percentages of PCE and the incidence of MN per 2000 PCE in both tissues in mice killed at 18 h were similar to the frequencies observed in mice terminated at 24 h. There were no significant differences in the percentage of PCE between control and the mice with or without UWBR exposure; the group mean values (+/- standard deviation) were in the range of 3.1+/-0.14 to 3.2+/-0.23 in peripheral blood, and 49.0+/-3.56 to 52.3+/-4.02 in bone marrow. The mean incidence of MN per 2000 PCE in control and in mice with or without UWBR exposure ranged from 7.7+/-2.00 to 9.7+/-2.54 in peripheral blood and 7.4+/-2.32 to 10.0+/-3.27 in bone marrow. Pairwise comparison of the data did not reveal statistically significant differences between the control and mice with or without UWBR exposure groups (excluding positive controls). CONCLUSION: Under the experimental conditions tested, there was no evidence for excess genotoxicity in peripheral blood or bone marrow cells of mice exposed to UWBR.

Ultra-wideband electromagnetic pulses and morphine-induced changes in nociception and activity in mice.

Mice were exposed to ultra-wideband (UWB) electromagnetic pulses averaging 99-105 kV/m peak amplitude, 0.97-1.03 ns duration, and 155-174 ps rise time, after intraperitoneal administration of saline or morphine sulfate. They were then tested for thermal nociception on a 50 degrees C surface and for spontaneous locomotor activity and its time profile over 5 min. Analysis of results showed no effect of UWB exposure on nociception and activity measures in CF-1 mice after 15-, 30-, or 45-min exposure to pulses at 600/s or after 30-min exposure to UWB pulses at 60/s. Similarly, no effect was seen in C57BL/6 mice after 30-min exposure to pulses at 60/s or 600/s. Although trends in morphine-modified measures seen with UWB pulse repetition frequency could be expected because of increased levels of low-frequency energy, no significant change was seen in normal or morphine-modified nociception or activity after UWB exposure. This indicated lack of effect of the UWB pulses used in these experiments on nervous system components, including endogenous opioids, involved in these behaviors.

Principles for Sustainable Governance of the Oceans

Pressures being exerted on the ocean ecosystems through overfishing, pollution, and environmental and climate change are increasing. Six core principles are proposed to guide governance and use of ocean resources and to promote sustainability. Examples of governance structures that embody these principles are given.

Ultra-wide band electromagnetic radiation does not affect UV-induced recombination and mutagenesis in yeast.

Cell samples of the yeast Saccharomyces cerevisiae were exposed to 100 J/m2 of 254 nm ultraviolet (UV) radiation followed by a 30 min treatment with ultra-wide band (UWB) electromagnetic pulses. The UWB pulses (101-104 kV/m, 1.0 ns width, 165 ps rise time) were applied at the repetition rates of 0 Hz (sham), 16 Hz, or 600 Hz. The effect of exposures was evaluated from the colony-forming ability of the cells on complete and selective media and the number of aberrant colonies. The experiments established no effect of UWB exposure on the UV-induced reciprocal and non-reciprocal recombination, mutagenesis, or cell survival.

Temporal bisection in rats: the effects of high-peak-power pulsed microwave irradiation.

The effects of high-peak-power, pulsed microwaves on a time perception and discrimination task were studied in rats. Exposures were performed with the TEMPO exposure system, which produces an 80 nanosecond pulse with peak-power levels in excess of 700 megawatts. The ability to expose animals to such fields within a controlled environment is unique. As determined by calorimetry, a maximal, whole-body-averaged, specific-absorption rate of 0.072 W/kg was produced. Thus exposures were well below a recommended SAR limit of 0.4 W/kg. Power levels of transmitted microwaves were varied over a 50 dB range to obtain ascending and descending dose-response functions for each of the behavioral measures. Measures of time perception, response bias, and total trials did not change with power level. Dose-response effects were observed for discriminability (ability to distinguish between durations), session time, and trial completions (null responses, failures to respond on a trial). Covarying sound and X-ray exposures produced by TEMPO did not reliably correlate with the observed microwave effects. The observation of repeatable dose-response effects on discriminability and null responses indicates that the microwave exposures were affecting cognitive function in the rats, particularly the decision-making process.

An allometric-autoregressive approach to poultry development.

1. The relative rates of development in a commercial layer and a broiler strain of chicken fed 180 or 230 g crude protein/kg were investigated by the use of the allometric-autoregressive model. Development was divided into pre-pubertal (0 to 8 weeks) and pubertal phases (8 to 22 weeks). 2. Significant strain effects were observed, at all ages, in rates of gain of live (L) body mass, carcass (C) mass, non-carcase (N) mass and empty-body (EB) mass as well as C, N and EB moisture, protein and fat content. 3. Significant dietary effects were observed in C and EB fat in the pre-pubertal phase and L mass, N protein and C fat in the subsequent pubertal period.

A microwave-transparent operant chamber for rats.

To study the operant behavior of rats in a microwave field, an operant chamber equipped with a response lever, stimulus lights, and a feeder was constructed using polycarbonate and fiber optic material. The chamber was connected to a personal computer located outside of the microwave chamber to program the schedules of reinforcement and to collect data. The operant chamber did not interfere with the microwave field and kept the subject's body parallel to the E-field for maximum absorption.

Calcium and magnesium content of skeletal muscle. Studies in subjects undergoing diagnostic testing for malignant hyperthermia.

Ca2+ and Mg2+ contents were measured and compared among three different malignant hyperthermia susceptible (MHS) diagnostic groups. No difference was found among mean values for Ca2+ content, whereas Mg2+ content was greater in MHS muscle. Variance of measured values was unequal and greatest among MHS muscles, suggesting a possible abnormal distribution associated with MHS. Although more muscle fibres with Ca2+ less than or equal to 12 mumol g-1 were observed in MHS muscle, this difference was not statistically significant. In contrast, non-parametric analysis showed that the population of Mg2+ values was significantly greater in the MHS muscle. This study suggests that the distribution of Ca2+ and Mg2+ values is different in MHS muscle as a result of unknown genetic factors associated with the disease.

Comparison of Ca++ uptake and spontaneous Ca++ release from sarcoplasmic reticulum vesicles isolated from muscle of malignant hyperthermia diagnostic patients.

Inward Ca++ transport and spontaneous Ca++ release activities were compared among sarcoplasmic reticulum (SR) membrane fractions isolated from human skeletal muscle in patients undergoing malignant hyperthermia (MH) diagnostic contracture testing. Two different membrane fractions were isolated, a heavy (8-12,000 X g) and light (12-48,000 X g) fraction, from each diagnostic subject. The rates of inward Ca++ transport were faster in light SR compared to heavy SR, but no statistically significant difference was observed among MH diagnostic groups. Spontaneous Ca++ release occurred at optimum Ca++ preload in all SR fractions and this preload did not differ among MH diagnostic groups. Optimal Ca++ preload for rate of spontaneously released Ca++ was greater in light SR compared to heavy SR. Similarly, rate of Ca++ release was faster in light SR than in heavy SR, but no difference in rate of spontaneously released Ca++ was observed among MH diagnostic groups. Amount of Ca++ released did not differ among SR fractions and it did not differ among diagnostic groups. In contrast to previous studies showing a defect in Ca++-induced Ca++ release, the mechanisms related to spontaneous Ca++ release and to oxalate-facilitated inward Ca++ transport, as measured in this study, appear to be normal in SR from human MH skeletal muscle.