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PubMed1968

Affinity site labeling of a mouse myeloma protein which binds dinitrophenyl ligands.

A mouse myeloma protein of the immunoglobulin A class, which specifically binds dinitrophenyl ligands, has been successfully affinity labeled (site-directed labeling) with several diazonium reagents, leading to inactivation of the combining sites. The labeling reagents reacted only with tyrosine residues of light chain origin. The 7S subunit of the myeloma protein appears to contain only one reactive site.

Animals↗
PubMed1973

Oxidative activation of Bacillus cereus spores.

A study was made of the activation of Bacillus cereus strain T spores by using the oxidizing agent sodium perborate. The degree of activation was measured with constant germination conditions by using L-alanine, inosine, adenosine, and L-alanine plus adenosine as germination stimulants. The germinal response following the various treatments was compared with the responses obtained with heat activation. It was concluded that the optimal time for activation with 30 mM sodium perborate at room temperature was about 4 hr. If the exposure time was greatly extended, the spores would germinate spontaneously. When the perborate treatment followed heat activation, the germinal response to L-alanine was stimulated, to inosine retarded and without apparent effect for adenosine or L-alanine plus adenosine. Results of experiments designed to demonstrate deactivation by slow oxidation showed that spores activated with sodium perborate were not deactivated by slow oxidation, whereas those activated by heat were. A deactivation study using mercaptoethanol as the deactivation agent showed that both methods of activation could be deactivated after a 24-hr exposure, but this deactivation was reversible by extending the exposure to mercaptoethanol. The results of heat-sensitivity studies revealed that about 70% of the sodium perborate-activated spores were heat sensitive after 60 min in a germination menstruum of L-alanine plus adenosine, whereas similarly treated heat-activated and nonactivated spores were about 99.99% heat sensitive, respectively.

Adenosine↗
PubMed1977

Effect of zinc ion on the hemolytic activity of thermostable direct hemolysin from Vibrio parahaemolyticus, streptolysin O, and Triton X-100.

The effect of Zn2+ on hemolysis induced by the thermostable direct hemolysin from Vibrio parahaemolyticus (vibriolysin) streptolysin O, and Triton X-100 was studied. We found that in certain buffers, such as tris(hydroxymethyl)-aminomethane-hydrochloride, boric acid-borax, and N-hydroxyethyl piperazine-N'-2-ethanesulfonic acid-sodium hydroxide, hemoglobins released from erythrocytes were easily precipitated by addition of Zn2+, thus resulting in a false inhibition of hemolysin by Zn2+ when hemolysis was assayed by measuring absorbance at 540 nm of released hemoglobins. Under experimental conditions in which hemoglobin was not precipitated, hemolysis induced by streptolysin O was inhibited by Zn2+, whereas that induced by vibriolysin and Triton X-100 was not. Thus, we concluded that the mode of inhibitory action of Zn2+ on hemolysis was not due to a reversible alteration in the state of the lipid bilayer of erythrocytes membranes as proposed by Avigad and Bernheimer (Infect. Immun. 13:1378-1381, 1976).

Buffers↗
PubMed1973

New method for the isolation of membranes from Mycoplasma gallisepticum.

Mycoplasma gallisepticum lysed readily in carbonate bicarbonate buffer at pH 9.2 to 10.5. The hemagglutination titer of the lysates was 2- to 16-fold greater than a cell suspension at the same protein concentration in buffered saline. Membranes prepared from cells lysed by this method at pH 10 were relatively free from cytoplasmic contaminants as shown by electron microscopy of thin sections. The membranes retained their hemagglutination activity, gave reactions in immunodiffusion tests identical to those obtained by osmotic lysis and sonic treatment, and showed a similar pattern of protein bands by polyacrylamide disk electrophoresis. When inoculated into rabbits, the membranes gave rise to antibodies active in growth-, metabolic- and hemagglutination-inhibition tests. On the average, membranes obtained by lysis at pH 10 contained 44% of the original cell protein. The method is simple, giving high yields of membranes, and may be adaptable to other mycoplasmas.

Animals↗
PubMed1981

Evaluation of the B-D urine culture kit.

Split samples of urine transported to the laboratory at 5 degrees C and in a boric acid-glycerol-sodium formate preservative (B-D Urine Culture Kit; Becton, Dickinson & Co.) were cultured immediately and, in the case of preserved urine, after 24 and 48 h of storage at 25 degrees C. Agreement between the results for cultures of specimens originally yielding greater than 10(5) colony-forming units (CFU) per ml and the results for urine preserved for 24 and 48 h was 85 and 71%, respectively. One-third of the specimens originally yielding 10(4) to 10(5) CFU per ml yielded less than 10(4) CFU per ml after 24 h of storage in preservative. Provided greater than 10(4) CFU per ml in specimens preserved for up to 24 h is regarded as equivalent to greater than 10(5) CFU per ml in original urine specimens, agreement of results was greater than 90%.

Bacteriological Techniques↗
PubMed1982

Urine culture transport tubes: effect of sample volume on bacterial toxicity of the preservative.

Stable bacterial counts in urine specimens before culture are necessary to assure the accurate diagnosis of urinary tract infections. Preservative-containing tubes are commercially available for urine transport. As these tube containers are not always filled to the manufacturer's specifications, we studied the effects of stabilizer formation with low urine volumes. The Sage Urine Culture Tube and the Becton-Dickinson Urine Culture Kit were evaluated by using 30 cultures diluted in urine to 10(5) colony-forming units per ml. Both tube types were injected with 1, 2, 3, and 4 to 5 ml (tube capacity) of urine containing each culture. Specimens were held at 22 degrees C and cultured at 0, 4, and 24 h. Colony counts were corrected for the dilution due to the preservative. The Becton-Dickinson Urine Culture Kits were toxic to Escherichia coli and Klebsiella pneumoniae in specimens containing up to 2 ml of urine, and the minimum usable amount of urine for reliable results was 3 ml. The Sage Urine Culture Tube maintained the number of bacteria in 1 to 4.5 ml of urine in 83% of the specimens. However, the Sage tube was toxic to E. coli when held for 24 h. Quantitative counts of enterococci tended to significantly increase in specimens that contained 2 ml or more of urine with either system. The limitation of preservative-containing tubes for urine transport need to be recognized in order to avoid false-positive and false-negative results.

Bacteria↗
PubMed1970

Deadly diapers.

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Boric Acids↗
PubMed1994

Limitations to the effectiveness of subcutaneous calcium solutions as a treatment for cows with milk fever.

Serum calcium concentrations were determined in 164 cases of milk fever (mostly Jersey cows) attended in the course of practice in a cold, wet winter rainfall area where the cows calved at pasture. The mean calcium concentration of the cows receiving no calcium therapy subcutaneously before sampling (1.0 mmol/litre) was significantly (P < 0.01) less than the mean of 1.4 mmol/litre for cows receiving 300 ml of 33.3 or 40 per cent calcium borogluconate solution, which was significantly (P < 0.01) less than the mean of 2.1 mmol/litre for those receiving 600 ml subcutaneously. Neither the rectal temperature of the cows nor the average time interval between the injection of the two bottles and blood sampling had any effect on the serum calcium of the latter cows, whereas the rectal temperature but not the time interval had a significant (P < 0.01) effect on the serum calcium of the cows receiving 300 ml; the lower their temperature, the lower the calcium concentration. The mean (+/- sd) interval between treatment by the herdsman and veterinary attention for the cows receiving 300 ml (4.8 +/- 4.9 hours was significantly (P < 0.01) shorter than for the cows receiving 600 ml (12.0 +/- 9.4 hours). At the time of sampling, 48 per cent of the cows receiving 600 ml of calcium borogluconate solution had a serum calcium concentration below 2.0 mmol/litre.

Animals↗
PubMed1997

Comparison of fluid and flunixin meglumine therapy in combination and individually in the treatment of toxic mastitis.

During a three-year study, 54 cows with toxic mastitis were allocated randomly to one of three treatment groups (A, B and C). Each cow was re-examined within 24 hours of the initial examination, and, during this time, group A received fluid therapy (45 liters of intravenous isotonic electrolyte solution) and flunixin meglumine (2000 mg), group B received fluid therapy only, and group C received flunixin meglumine only. In addition all the cases were treated with parenteral and intramammary tetracyclines, oxytocin and calcium boroglucoanate. There was no significant difference in the rate of survival between the treatment groups and 29 of the cows (53.7 per cent, 95 per cent confidence interval of 39 to 67 per cent) survived.

Animals↗