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Biomedical subjects

C J Knowles

Publications and source records attributed to C J Knowles.

At least 37 records · Page 2Linked to original sources

Effects of unilateral and bilateral auricular transcutaneous electrical nerve stimulation on cutaneous pain threshold.

This study compared the effects of unilateral and bilateral auricular transcutaneous electrical nerve stimulation on cutaneous pain threshold. Auricular acupuncture points were stimulated with low frequency, high intensity TENS for 45 seconds. Sixty healthy, adult subjects were assigned randomly to one of two treatment groups or to a control group. The two treatment groups received low frequency, high intensity TENS either unilaterally or bilaterally. The control group did not receive auricular stimulation. Experimental pain threshold at the left wrist was determined with a painful stimulus before and after auricular stimulation. Both unilateral and bilateral auricular stimulation groups exhibited a significant increase (p less than .05) in experimental pain threshold, but the control group did not. The mean change values between the unilateral and bilateral stimulation groups were not statistically different. These results suggest that both unilateral and bilateral auricular TENS can increase pain threshold.

Adult↗

Ethylene formation by cell-free extracts of Escherichia coli.

The pathway leading to the formation of ethylene as a secondary metabolite from methionine by Escherichia coli strain B SPAO has been investigated. Methionine was converted to 2-oxo-4-methylthiobutyric acid (KMBA) by a soluble transaminase enzyme. 2-Hydroxy-4-methylthiobutyric acid (HMBA) was also a product, but is probably not an intermediate in the ethylene-forming pathway. KMBA was converted to ethylene, methanethiol and probably carbon dioxide by a soluble enzyme system requiring the presence of NAD(P)H, Fe3+ chelated to EDTA, and oxygen. In the absence of added NAD(P)H, ethylene formation by cell-free extracts from KMBA was stimulated by glucose. The transaminase enzyme may allow the amino group to be salvaged from methionine as a source of nitrogen for growth. As in the plant system, ethylene produced by E. coli was derived from the C-3 and C-4 atoms of methionine, but the pathway of formation was different. It seems possible that ethylene production by bacteria might generally occur via the route seen in E. coli.

Escherichia coli↗

Ethylene formation by cultures of Escherichia coli.

Growth of Escherichia coli strain B SPAO on a medium containing glucose, NH4Cl and methionine resulted in production of ethylene into the culture headspace. When methionine was excluded from the medium there was little formation of ethylene. Ethylene formation in methionine-containing medium occurred for a brief period at the end of exponential growth. Ethylene formation was stimulated by increasing the medium concentration of Fe3+ when it was chelated to EDTA. Lowering the medium phosphate concentration also appeared to stimulate ethylene formation. Ethylene formation was inhibited in cultures where NH4Cl remained in the stationary phase. Synthesis of the ethylene-forming enzyme system was determined by harvesting bacteria at various stages of growth and assaying the capacity of the bacteria to form ethylene from methionine. Ethylene forming capacity was greatest in cultures harvested immediately before and during the period of optimal ethylene formation. It is concluded that ethylene production by E. coli exhibits the typical properties of secondary metabolism.

Ammonium Chloride↗

Synthesis of catalase by "Streptococcus faecalis subsp. zymogenes".

"Streptococcus faecalis subsp. zymogenes" was grown aerobically and anaerobically with glycerol as the source of carbon, and in the presence and absence of haematin. Catalase activity was found only during aerobic growth in the presence of haematin. The rate of appearance of catalase activity was measured on (a) addition of haematin to haematin-less aerobic cultures, and (b) aeration of haematin-containing anaerobic cultures in the presence or absence of chloramphenicol. These experiments suggested that apocatalase synthesis was induced by aeration and was not dependent on the presence of haematin in the growth medium. The binding of haem to apocatalase was oxygen dependent.

Aerobiosis↗

Isolation and growth of a Pseudomonas species that utilizes cyanide as a source of nitrogen.

A simple method of isolating bacteria that utilize cyanide as a source of nitrogen for growth has been developed. This involved supplying hydrogen cyanide as a vapour to glucose-containing minimal-salts agar plates. The bacteria isolated were Gram-negative, oxidase-positive rods producing a fluorescent green pigment and were tentatively identified as strains of Pseudomonas fluorescens. Three organisms were studied further and shown to be P. fluorescens biotype II. One of these (NCIB 11764) was grown in a glucose-containing fed-batch culture with either NH4Cl or KCN as the limiting nutrient. Cyanide-grown bacteria produced stoichiometric amounts of ammonia from cyanide when pulsed with cyanide under aerobic conditions. Stimulation of oxygen uptake was seen on addition of cyanide to suspensions of cyanide-grown but not ammonia-grown bacteria.

Ammonium Chloride↗

Growth of Streptococcus faecalis var. zymogenes on glycerol: the effect of aerobic and anaerobic growth in the presence and absence of haematin on enzyme synthesis.

Streptococcus faecalis var. zymogenes was grown aerobically and anaerobically in the presence and absence of haematin, with glycerol as the carbon and energy source. Aerobic growth was stimulated by the inclusion of haematin in the medium but fumarate had no effect on growth. The bacterium was unable to grow anaerobically on glycerol unless fumarate was present; haematin had no effect on growth. NADH oxidase activity, which catalysed the oxidation of NADH + H+ to form H2O rather than H2O2, was found in the soluble fraction and was induced by aerobic growth but partially repressed when haematin was present in the medium. In contrast, a particulate NADH oxidase, which was sensitive to inhibition by antimycin A and 2-heptyl-4-hydroxyquinoline N-oxide, was induced by aerobic growth in the presence of haematin. NADH peroxidase was massively induced by aerobic growth, whereas more lactate dehydrogenase activity was found in anaerobically grown bacteria. Catalase was formed only during aerobic growth in the presence of haematin.

Aerobiosis↗

The effect of haematin and catalase on Streptococcus faecalis var. zymogenes growing on glycerol.

Streptococcus faecalis var. zymogenes was grown aerobically on a complex medium containing glycerol as the carbon source. Addition of hematin or bovine liver catalase to the growth medium resulted in a small increment in growth yield. Suspensions of bacteria that had been grown in the presence of haematin or catalase, respectively, translocated 0.83 to 1.98 and 1.33 to 2.53 protons per oxygen atom consumed in glycerol oxidation. Bacterial grown without haematin or catalase had nil or little respiratory-induced proton translocation during glycerol oxidation. Inclusion of haematin in the growth medium caused the bacterium to form a cyanide- and azide-sensitive catalase. Superoxide dismutase activity was similar whether or not haematin was added to the growth medium.

Catalase↗

AMP metabolism in the marine bacterium Beneckea natriegens.

The catabolism of AMP by preparations from Beneckea natriegens has been reexamined. In the absence of ATP, cell-free extracts catabolized AMP via adenosine to inosine. When ATP was present, adenylate kinase converted AMP to ADP, lowering the rate of AMP catabolism. Particle-free supernatants (225,000 x g) metabolized AMP alone slowly, but adenylate kinase was active when ATP was added. Washed particulate fractions contained AMP nucleotidase activity which converted AMP to adenosine; in the presence of ATP, adenosine formation was reduced by residual adenylate kinase associated with the particulate fraction. IMP was not detected as a metabolite in these experiments.

5'-Nucleotidase↗

The regulatory effects of growth rate and cyclic AMP levels on carbon catabolism and respiration in Escherichia coli K-12.

Cyclic AMP levels in glucose and succinate-limited and ammonia-limited glucose-containing continuous cultures of Escherichia coli were measured at different bacterial growth rates. Intracellular cyclic AMP concentrations were fairly constant (about 5 micrometer) at all dilution rates used when glucose was limiting. In ammonia-limited glucose cultures the cyclic AMP content was much lower (about 0.3 micrometer). In succinate-limited cultures cyclic AMP levels fell from 2.7 to 0.8 micrometer as dilution rate increased from 0.05 to 0.4 h-1. The effects of cyclic AMP on respiratory and carbon catabolic enzyme levels were studied. There was no indication of a direct cyclic AMP involvement in the regulation of these cellular functions. It seems more likely that the variations in enzyme levels observed resulted from variation of the specific growth rate of cultures.

Ammonia↗

Transhydrogenase activity in the marine bacterium Beneckea natriegens.

The marine bacterium, Beneckea natriegens, which has previously been reported not to form transhydrogenase, has been shown to synthesize a soluble energy-independent transhydrogenase (NADPH:NADP+ oxidoreductase, EC 1.6.1.1), though no energy-linked activity could be detected. The transhydrogenase is induced maximally in stationary phase cells and its formation is 70-90% repressed by raising the medium phosphate level from 0.33 to 3.3 mM. The enzyme is inhibited by arsenate, inorganic ortho- and pyrophosphate and by a range of organic phosphate-containing compounds, including 2'-AMP, which is an activator of several bacterial transhydrogenases.

Bacteria↗

Adenylate energy charge during batch culture of Beneckea natriegens.

The value of the adenylate energy charge, i.e. ([ATP] + 1/2[ADP])/([ATP] + [ADP] + [AMP]), during batch culture of Beneckea natriegens remained relatively constant during the exponential and early stationary phases of the growth cycle. During exponential growth the intracellular ATP content remained constant, the amount of ATP in the culture increasing proportionally with growth; these conditions were unaltered during growth in the presence of added cyclic AMP. On cessation of growth, significant variation in bacterial ATP content was observed depending on whether growth of the cultures terminated due to exhaustion of carbon or nitrogen from the medium, and on the presence or absence of added cyclic AMP.

Adenine Nucleotides↗

The respiratory system of Chromobacterium violaceum grown under conditions of high and low cyanide evolution.

The particulate fraction of disrupted Chromobacterium violaceum grown under cyanide-evolving conditions was unable to oxidize ascorbate plus N,N,N',N'-tetra-methyl-p-phenylenediamine (TMPD), but oxidized NADH and succinate by a linear respiratory pathway which was very resistant to inhibition by cyanide. When the bacteria were grown under conditions where little cyanide evolution occurred, particulate fractions developed the ability to oxidize ascorbate-TMPD by a pathway highly sensitive to cyanide inhibition; respiratory activity with NADH and succinate proceeded via both the cyanide-sensitive and-resistant pathways. Studies with respiratory inhibitors, and the cytochrome compositions of the fractions derived from cultures grown under both conditions, are presented. A soluble, carbon monoxide-binding cytochrome c was found, and this appears similar to those found recently in Beneckea natiegens, methylotrophic bacteria and the marine pseudomonad B16.

Antimycin A↗