Detergent protease exposure and respiratory disease: case-referent analysis of a retrospective cohort.

OBJECTIVES: To examine the relationship between protease exposure and respiratory disease in a cohort of detergent enzyme manufacturers. METHODS: Case-referent analysis of a cohort of employees working in a European detergent factory between 1989 and 2002. Cases with new lower or upper respiratory disease were ascertained by examination of occupational health records and matched to referents on date of first employment. Personal exposures to airborne detergent protease were estimated, using a job exposure matrix, from >12,000 measurements taken in the factory during the period of study. RESULTS: We found clear, monotonic relationships between estimated protease exposure and both lower and upper respiratory disease. After control for age, sex and smoking, the odds ratio of lower respiratory disease was significantly elevated (1.98, 95% CI 1.04 to 3.79) in those employees working in jobs in the highest quartile of protease exposure (geometric mean 7.9 ng x m(-3)). For employees with upper respiratory disease, the risk was significantly elevated at a lower level of estimated protease exposure (geometric mean 2.3 ng x m(-3)). CONCLUSIONS: These findings provide strong evidence of an association between detergent enzyme exposure and the development of respiratory disease in an occupational setting. Using the routinely collected information on specific sensitisation and the close attention to workplace exposures that are characteristic of this industry, it should be possible to derive meaningful occupational exposure standards for most detergent enzymes.

The antidiabetic activity of the herbal preparation ADD-199 in mice: a comparative study with two oral hypoglycaemic drugs.

The antidiabetic and antioxidant effects of the herbal preparation ADD-199 were investigated in STZ-induced diabetic C(3)H mice and results were compared with two allopathic hypoglycaemic drugs, glibenclamide and metformin. Plasma glucose, insulin and lipids as well as liver glycogen, lipids and lipid peroxidation were measured following treatment for 8 weeks. The results indicated that plasma insulin levels in normal controls at termination were about 76 micromol/L compared to trace levels in untreated diabetic mice. Glibenclamide and ADD-199 increased insulin levels in diabetic mice up to 70% of levels in untreated non-diabetic mice whilst metformin had no effect. Basal plasma glucose levels in diabetic controls (18.8 mM) were reduced to 14.0 mM by 100 mg/kg ADD-199 in <2 weeks compared to 4 and 6 weeks for glibenclamide and metformin, respectively. This hypoglycaemic effect of ADD-199 appeared to be associated with the alkaloidal content of the extract. Treatment with ADD-199 or the hypoglycaemic agents reversed the observed elevation in plasma lipids but increased hepatic glycogen, triacylglycerol and cholesterol levels. Treatment also increased glucose uptake by isolated diaphragms and attenuated hepatic lipid peroxidation. These antihyperglycaemic and antioxidant actions of ADD-199 at a dose of 100mg/kg/day are comparable to those of the maximum daily therapeutic doses of glibenclamide (0.25 mg/kg) and metformin (50 mg/kg). These could explain the basis for use of this plant extract to manage diabetes mellitus (DM).

Oocyte maturation.

The oocyte is dependent on granulosa cells to provide nutrients and regulatory signals. Granulosa cells must be at the appropriate stage of differentiation to initiate these signals and transmit them to the oocyte. Studies have shown that in vitro-matured oocytes from follicles in early stages of atresia are more competent to support embryonic development than those from actively growing follicles. The acquisition of developmental competence appears to occur prior to in vitro maturation and can be induced by gonadotropin-free coasting in vivo or postmortem ovary incubation in vitro. The acquisition of developmental competence is probably a common signaling or differentiation pathway that occurs in the oocyte and/or associated granulosa regardless of whether the oocyte is destined to ovulate or degenerate. Early follicle atresia is the visually discernible characteristic in vitro that is associated with increased developmental potential.

Production of embryos from in vitro-matured primary human oocytes.

OBJECTIVE: To determine the factors that influence the number and quality of embryos produced from primary oocytes collected from untreated regularly ovulating and irregular or anovulatory polycystic women. DESIGN: A direct comparison between two patient groups whose oocytes were matured in vitro and a comparison of the embryo development of in vitro-matured oocytes from untreated patients with in vivo-matured oocytes of superovulated IVF-ET patients obtained during the same period. SETTING: The Monash IVF Clinic, involving patients who expressed the desire to avoid super-ovulation with fertility drugs. MAIN OUTCOME MEASURES: The completion of nuclear maturation of oocytes after 36 or 48 hours culture, fertilization in vitro, and embryo development ratio. RESULTS: Oocytes from regular cycling patients matured and fertilized at significantly higher rates than irregular cycling and anovulatory women and their embryos had significantly higher mean embryo development ratio. The mean embryo development ratio of embryos of regular cycling patients was similar to superovulated IVF patients but irregular cycling and anovulatory patients had a significantly lower embryo development ratio. Culture of oocytes for 48 hours increased maturation of oocytes from 57% to 82% but did not affect fertilization or cleavage rates. Embryo development was not affected significantly by the grade of follicular cell cover of oocytes. CONCLUSIONS: The developmental capability of primary oocytes is higher in regular cycling women than in irregular cycling and anovulatory women with polycystic ovary disease.

Temporally-specific modification of myelinated axon excitability in vitro following a single ultrasound pulse.

Single, short-duration, low-energy pulses of ultrasound were found to elicit distinct modifications of the electrical excitability of myelinated frog sciatic nerve in vitro in a window extending 40-50 ms after pulse termination. These modifications include both enhancement and suppression of relative excitability, the sequence of which generally follows one of two distinct temporal response patterns. The ultrasound pulses were focused, 2-7 MHz, of 500-microseconds duration, and of peak intensities of 100-800 W/cm2. Total absorbed pulse energies were generally less than 100 mJ/g, corresponding to local temperature rises of the nerve trunk of no more than 0.025 degrees C per pulse, thereby precluding bulk heating as a basis of this effect. The observed effects cannot be elicited using either a subthreshold square wave or RF electrical prestimulus, suggesting a unique form of receptivity of the nerve trunk to mechanical perturbation. We present evidence that the low-frequency radiation pressure transient accompanying the envelope of the acoustic pulse is the active parameter in this phenomenon, and postulate that it may act by the gating of stretch-sensitive channels, which have been recently reported in a variety of cell membranes. These results may demonstrate that stretch-sensitive channels in neural membrane can serve to functionally modulate neuro-electric signals normally mediated by voltage-dependent channels, a finding which could suggest new clinical applications of high peak-power, low-total-energy pulsed ultrasound.

Real-time multichannel computerized electrogastrograph.

The purpose of this study was to develop a real-time multichannel computerized electrogastrograph (EGG) to measure and analyze electrical signals from the human abdominal surface. A soft-contact matrix composed of 25 cutaneous electrodes is embedded evenly in a latex mat. The mat can be firmly attached to the abdominal surface by drawing a vacuum between the matrix and the stomach. Twenty-five high-amplification filter/amplifiers provide a high signal-to-noise ratio and flat amplitude response for a signal between 0.02 and 0.12 Hz (1.2-7.2 cpm). The computer program provides waveform and frequency analysis for any chosen channel and mapping analyses for all 25 channels. A two-dimensional propagation exploration program was also developed. Using four different mapping analysis program subroutines, the optimal points for analyzing the EGG signals can be reliably found and variability of these locations can be observed easily. Results show differences in the EGG mappings of normal and abnormal subjects.

Formate species in the low-temperature oxidation of dimethyl ether.

The oxidation of dimethyl ether (DME, 340 ppm in 10% O2) has been studied experimentally in an atmospheric pressure laminar flow reactor in the temperature range from 240 degrees C to 700 degrees C for residence times in the range 2-4 s. The influence of nitric oxide additions up to 620 ppm to the feed gases has also been investigated. Products of reaction were determined by FTIR. In the absence of NO, reaction is first detected at about 260 degrees C. The products in the low-temperature region include formaldehyde (HCHO), and formic acid (HCOOH). The addition of NO leads to the appearance of methyl formate (CH3OCHO). While the overall behaviour of the system can be explained qualitatively in terms of typical low-temperature hydrocarbon ignition, recently published chemical kinetic models for DME ignition do not allow for the formation of these formate species. We find no experimental evidence for the formation of hydroperoxymethyl formate (HPMF, HOOCH2OCHO) which is predicted by the models to be a significant stable intermediate at temperatures below 350 degrees C. Since both formic acid and methyl formate have potentially harmful health effects, these observations may have significant implications for use of DME as a diesel fuel.

The effects of the early uterine environment on the subsequent development of embryo and fetus.

Synchrony between the embryo and the uterine endometrium is essential for the establishment of pregnancy and birth in people and livestock. When asynchronous conditions occur a variety of complication result that include failure of the embryo to implant, early embryonic mortality, retarded development and growth, and accelerated development and growth. These complications all appear to be induced within the first week of embryo development and not withstanding the immediate endpoint of large or small size at birth, may alter the course of development throughout the life of the animal. Progesterone appears to play a causative role in establishing the abnormal growth of the fetus by decelerating or accelerating embryonic development. This may act through increasing the transport of blood born growth factors into the uterine lumen or by stimulating the release of growth factors from the endometrium directly. It can not be ruled out that progesterone mediated abundance of, or absence of, appropriate nutrition may bring about the same lifelong outcome. In vitro culture situations that include serum and/or co-culture can also bring about these abnormalities of growth. It is hypothesized that exposure to growth factors "out of phase" may result in an irreversible induction of abnormal development. The described abnormalities that occur in sheep and cattle have not yet been described for children resulting from IVF.

Reducing the amount of cytoplasm available for early embryonic development decreases the quality but not quantity of embryos produced by in vitro fertilization and nuclear transplantation.

The effect of reducing the amount of cytoplasm available for early embryonic development was investigated in embryos produced by in vitro fertilization (IVF) and nuclear transplantation. In Experiment 1, approximately 1/2 or 1/20 of the cytoplasm was removed from bovine embryos at the pronuclear-stage of development. The percentage of embryos developing to the compact morula or blastocyst stage was significantly higher in non-manipulated controls (26%) than in embryos with 1/20 of the cytoplasm removed (16%), and those with 1/2 of the cytoplasm removed (10%; P < 0.05). There was also a significant difference in the average number of cells between blastocysts in which 1/20 of their cytoplasm was removed (67), those with 1/2 of their cytoplasm removed (55), and nonmanipulated controls (77; P < 0.05). In Experiment 2, nuclear transfer embryos were produced in which approximately 1/2 or 1/20 of the cytoplasm was removed during oocyte enucleation. The percentage of embryos developing to the blastocyst stage was 17% for both groups of nuclear transfer embryos compared to 44% for control embryos (P < 0.05). The mean number of cells in blastocysts produced by nuclear transfer in which 1/20 of the cytoplasm was removed during oocyte enucleation (61) was no different than that in control embryos (66), but significantly higher than the mean number of cells in blastocysts produced by nuclear transfer in which 1/2 of the cytoplasm was removed (42; P < 0.05). There was no indication that altering the amount of cytoplasm available for early embryonic development of IVF embryos affected the timing of differentiation events, including those of embryo compaction and blastocyst formation.

Nuclear transplantation in bovine embryos.

This study was conducted to develop a method for transplanting nuclei in bovine embryos and to test the development of several stages of donor nuclei transplanted to enucleated pronuclear recipient embryos. Pronuclear embryos were centrifuged to reveal nuclei. Nuclei were removed without penetrating the plasma membrane as membrane-bound karyoplasts, and were inserted into enucleated zygotes by electrically induced cell fusion. The highest rate of fusion (79%) occurred in Zimmerman Cell Fusion medium at 100 V for 20 to 40 microseconds with the fusion membranes oriented parallel to the electrodes. The effect of nuclear transplantation on development was tested in pronuclear embryos in which nuclei were removed and reinserted and the embryos were then transferred to sheep oviducts for 5 d. Of the intact nuclear transplant embryos recovered, 5/29 (17%) developed to morulae or blastocysts compared with 11/30 (37%) of the non-manipulated embryos. Two nuclear transplant embryos were transferred to a recipient cow, and both developed to normal offspring. When nuclei from two-, four-, or eight-cell embryos were transplanted to pronuclear recipient embryos, no development was observed.

Isolation of mastitis pathogens from quarters of cows recently treated with antimicrobial agents.

Milk samples for aerobic culturing were collected from 72 quarters of cows with clinical mastitis that had been nonresponsive to antimicrobial intramammary treatment. All samples were collected within 24 hours of the most recent treatment. Mastitis pathogens were isolated from 56 samples (78%).

Radio-microwave interactions with biological materials.

There are a variety of ways in which a microwave field can affect a biological system. The best understood are those associated with heating, which, in turn, lead to changes in chemical reaction rates and current flows. At high levels, forces associated with the field gradients have been observed, as well as nonlinearities, including the destruction of membranes. At low levels, we discuss the possibility of quantum effects and some limits set by noise.