Trichloroacetic acid infections

A 47-year-old woman undergoing face peeling with 35% trichloroacetic acid developed left eye burning sensation, excessive tearing, marked conjunctival injection, conjunctival infection, and mild inferior superficial punctuate keratitis involving 25% of the cornea [32],... 

To allow for Inevitable differences In Infection levels between experiments, transfer was expressed relative to the incorporation of HJ methionine into trichloroacetic acid insoluble material (= protein), which provided a measure of mildew growth. This technique only accounted for radioactivity In conidla and surface mycelium, not haustorla, but significant differences In the transfer of triadlmenol were apparent between strains (Table III). These differences were not, however, correlated with triadlmenol sensitivity, so that if reduced accumulation is involved in DMI resistance in barley powdery mildew, it must not operate in all resistant strains. In the same experiments we looked at the possibility that detoxification of triadlmenol might account for resistance, but were unable to find any evidence of triadlmenol metabolites in the six strains examined. 

Protein synthesis was determined in the non-infected cells used as control and the HSV-1 and/or VSV infected cells, in order to obtain quantitative data on the toxicity and antiviral activity of the compounds assayed. Thus, 0.5 free medium methionine and 1 pCi/ml of methinione 32S were added to die cells and incubated at 37°C for 1 h. Thereafter the medium was siphoned off and the cells were washed with saline phosphate buffer and precipitated with 5% trichloroacetic acid (TCA). After 5 min, the acid was eliminated and the cells were washed twice with 96% ethanol,dried under an infrared light and dissolved in NaOH 0.1N with DSS. A liquid scintillation counter was used for the recount. The results showed no antiviral activity for die dihydro-f -agarofuran sesquiterpenes. 

Figure 1. Time course of protein and viral RHA synthesis in HeLa cells infected with EMC. A culture at 4x10 cells/ml in MEM medium, 59 fetal calf serum, 2mM glutamine and 14 mM HEPES-KOH, pH 7 2, was infected with JOOpfu/cell of EMC. To measure protein synthesis, 0.1 ml of the culture were added at the indicated times to 0.9 ml of methionine-free medium containing 10 0 /ml of 35S )-methionine and incubated 20 min. To measure EHA synthesis, 0.2 ml of the culture were added to 0.8 ml of medium containing 6.25 M g /ml of actinomycin D and incubated 5 min 10 ixCi of ( 3H )-uridine were then added for 1 5 min. The incubations were stopped by adding 10 ml of cold spinner salts solution and the cells collected by centrifugation. The pellet was dissolved in 1% Ha-dodecyl sulphate and precipitated with 5% trichloroacetic acid for counting.

Figure 6. Electrophoretic analysis of proteins associated with native 40 S ribosomal subunits of control and EMC-infected HeLa cells. Extracts were prepared from mock-infected and infected cells as described in Figure 2 0.5 ml of extract from control cells (b) or from cells infected 2 hr (a) or 3 hr (c) with EMC were layered on 15-50% sucrose gradients in 30 mM KC), 2mM Mg(OAc)p, 20 mM HHPES-KOH, pH 7 4i 1 mM dithiothreitol, and centrifuged 17 hr at 50f000 rpm. Fractions corresponding to the 40 S peak were combined, precipitated with 10% trichloroacetic acid, washed with acetone/ether (2/3) and ether, dissolved in sample buffer and fractionated on 12,5% polyacrylamide gels as previously described (14) The gels were stained with Goomassie Blue. Markers were run in parallel to assign M to the bands indicated by arrows.

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