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Biomedical subjects

A J Carlisle

Publications and source records attributed to A J Carlisle.

11 recordsLinked to original sources

Exercise and outdoor ambient air pollution.

OBJECTIVES: To establish by literature survey: (a) levels at which air pollutants are considered damaging to human health and to exercisers in particular; (b) the current ambient levels experienced in the United Kingdom; (c) whether athletes are especially at risk. METHODS: Six major urban air pollutants were examined: carbon monoxide (CO); nitrogen oxides (NO(X)); ozone (O(3)); particulate matter (PM(10)); sulphur dioxide (SO(2)); volatile organic compounds (VOCs). RESULTS: CO is detrimental to athletic performance. NO(2) is of concern to human health, but outdoor levels are low. O(3) poses a potentially serious risk to exercising athletes. Decrements in lung function result from exposure, and there is evidence that athletic performance may be affected. Detrimental effects may occur at low ambient levels, but there is no scientific consensus on this matter. PM(10) is causing concern in the scientific community. Blood lead accumulation during exercise indicates that personal exposure to toxic compounds associated with PM(10) may be magnified. Generally, outdoor ambient levels of SO(2) are too low to cause a problem to the athlete, except the asthmatic athlete. The few studies on exposure of exercisers to VOCs are reviewed. CONCLUSIONS: Athletes and exercisers should avoid exercising by the road side even though levels of the more noxious air pollutants have been controlled in the United Kingdom. O(3) is particularly damaging to athletes; it reaches its highest concentrations on hot bright days in rural areas.

Air Pollutants↗

Development of a prostate cDNA microarray and statistical gene expression analysis package.

A cDNA microarray comprising 5184 different cDNAs spotted onto nylon membrane filters was developed for prostate gene expression studies. The clones used for arraying were identified by cluster analysis of > 35 000 prostate cDNA library-derived expressed sequence tags (ESTs) present in the dbEST database maintained by the National Center for Biotechnology Information. Total RNA from two cell lines, prostate line 8.4 and melanoma line UACC903, was used to make radiolabeled probe for filter hybridizations. The absolute intensity of each individual cDNA spot was determined by phosphorimager scanning and evaluated by a bioinformatics package developed specifically for analysis of cDNA microarray experimentation. Results indicated 89% of the genes showed intensity levels above background in prostate cells compared with only 28% in melanoma cells. Replicate probe preparations yielded results with correlation values ranging from r = 0.90 to 0.93 and coefficient of variation ranging from 16 to 28%. Findings indicate that among others, the keratin 5 and vimentin genes were differentially expressed between these two divergent cell lines. Follow-up northern blot analysis verified these two expression changes, thereby demonstrating the reliability of this system. We report the development of a cDNA microarray system that is sensitive and reliable, demonstrates a low degree of variability, and is capable of determining verifiable gene expression differences between two distinct human cell lines. This system will prove useful for differential gene expression analysis in prostate-derived cells and tissue.

DNA, Complementary↗

Skeletal muscle damage following halothane anaesthesia in the domestic fowl: plasma biochemical responses.

The effects of a short period (10 minutes) of halothane anaesthesia upon skeletal muscle have been examined in broiler chickens. Integrity of the muscle membrane was assessed by measurement of plasma activities of intracellular enzymes. Creatine kinase activity was greatly increased during the first 12 hours post-anaesthesia and remained elevated for at least 48 hours. Lactate dehydrogenase activity exhibited a similar pattern which was less pronounced. Halothane anaesthesia induced transient hypercapnic acidosis and hypokalemia and increased corticosterone secretion but there was no significant effect upon deep body temperature. It is proposed that halothane may act directly upon skeletal muscle, perhaps influencing intracellular calcium homeostasis, to alter membrane permeability and increase enzyme efflux reflecting a degree of post-anaesthetic muscle damage. The transient changes in electrolyte and acid-base balance may contribute to these effects. Precautions should be taken to minimise the incidence or extent of halothane induced myopathy in birds particularly in rapidly growing broiler chickens where susceptibility may be increased.

Anesthesia↗

An improved method for construction of directionally cloned cDNA libraries from microdissected cells.

Here, we developed an improved method for constructing microdissected cDNA libraries, based on strand-switching properties of reverse transcriptase, followed by PCR amplification with primers to mediate unidirectional insert cloning. Using RNA from microdissected ovarian carcinoma cells, we constructed a cDNA library consisting of 1.3 x 10(6) unidirectional recombinants with an average insert size of 500 bp. Single-pass sequencing of 100 clones with the T7 primer revealed 89 inserts derived from known genes, anonymous expressed sequence tags (ESTs), or novel sequences. Among these clones were known genes and ESTs previously found in cDNA libraries from bulk ovarian tissue RNA, sequences seen for the first time in an ovarian-derived library, and novel sequences not previously seen in any cDNA library. These results demonstrate a methodology for constructing quality cDNA libraries that are cloned in a unidirectional fashion, are complex and diverse, and reflect the tissue of origin.

Actins↗

The effects of chronic exposure to elevated environmental temperature on intestinal morphology and nutrient absorption in the domestic fowl (Gallus domesticus).

1. Exposure of growing broiler chickens to elevated environmental temperature (35 degrees C) for two weeks, markedly reduced food intake (29%) and growth rate (37%) compared to birds maintained at 22 degrees C. 2. These changes in growth were accompanied by increased in vivo jejunal uptakes of galactose (36%) and methionine (50%) measured per unit intestinal dry weight. 3. Both the electrogenic (phloridzin sensitive) and non-electrogenic (phloridzin insensitive) components of galactose absorption were increased by 24 and 52% respectively during the chronic heat stress. 4. The size of the absorptive compartment may be reduced by the heat stress as reflected by decreased villus heights (19%) and wet (26%) and dry (31%) weights per unit length of jejunum. 5. It is suggested that the changes in hexose and amino acid during chronic exposure to elevated ambient temperature may reflect adaptations to optimise nutrient absorption in the face of reduced nutrition and decreases in the size of the absorptive compartment. A functional hypothyroidism (plasma luminal T3 decreased by 66%) associated with heat stress may contribute to the observed alterations in jejunal structure and function.

Animals↗

Plasma zinc as an index of vitellogenin production and reproductive status in the domestic fowl.

1. A technique is described for the accurate determination of circulating vitellogenin concentrations by measurement of plasma zinc concentrations before and after selective precipitation of lipoproteins in the domestic fowl. 2. Vitellogenin zinc concentrations exhibit a high degree of correlation with those of another major egg yolk precursor, very low density lipoprotein (VLDL) in immature female birds, during developing and maximum egg production in mature birds and following cessation of egg laying. 3. Mature female patterns of plasma vitellogenin and VLDL were induced by injection of oestradiol 17 beta (5 mg/kg body weight per day for 3 days) into adult cockerels. 4. It is suggested that measurement of plasma zinc provides a simple and accurate technique for the estimation of vitellogenin production and reproductive status in the domestic fowl and that this may be applied to other oviparous vertebrates.

Aging↗

Comparison of the stressfulness of harvesting broiler chickens by machine and by hand.

The procedures of manual and machine catching acted as short-term stressors, and induced a state of fear in 8-week-old broiler chickens. The heart rate of birds caught by both methods rose to similar high values but that of birds caught by machine returned to near normal rates more quickly, suggesting that they were less stressed. The duration of tonic immobility, a response which increases with fearfulness, was much longer in manually-caught birds. These results suggest that stress could be reduced and welfare improved by catching and picking up broiler chickens by a carefully designed machine, rather than by hand.

Animals↗