Ethylene inhibits aflatoxin biosynthesis in Aspergillus parasiticus grown on peanuts.

The filamentous fungi Aspergillus parasiticus and Aspergillus flavus synthesize aflatoxins when they grow on a variety of susceptible food and feed crops. These mycotoxins are among the most carcinogenic naturally occurring compounds known and they pose significant health risks to humans and animals. We previously demonstrated that ethylene and CO2 act alone and together to reduce aflatoxin synthesis by A. parasiticus grown on laboratory media. To demonstrate the potential efficacy of treatment of stored seeds and grains with these gases, we tested ethylene and CO2 for ability to inhibit aflatoxin accumulation on Georgia Green peanuts stored for up to 5 days. We demonstrated an inverse relationship between A. parasiticus spore inoculum size and the level of toxin accumulation. We showed that ethylene inhibits aflatoxin synthesis in a dose-dependent manner on peanuts; CO2 also inhibits aflatoxin synthesis over a narrow dose range. Treatments had no discernable effect on mold growth. These observations support further exploration of this technology to reduce aflatoxin contamination of susceptible crops in the field and during storage.

Ethylene modulates development and toxin biosynthesis in aspergillus possibly via an ethylene sensor-mediated signaling pathway.

Ethylene, a biologically active natural compound, inhibited aflatoxin accumulation by Aspergillus parasiticus on a solid growth medium in a dose-dependent manner at concentrations of 0.1 to 150 ppm. The activity of the nor-1 promoter (an early aflatoxin gene) was reduced to nondetectable levels by similar quantities of ethylene, suggesting that the inhibitory effect on toxin synthesis occurred, at least in part, at the level of transcription. The inhibitory effect of ethylene on aflatoxin accumulation was also observed when A. parasiticus was grown on raw peanuts. Under similar growth conditions and doses, ethylene strongly inhibited development of asci and ascospores in Aspergillus nidulans, with no detectable effect on Hülle cell formation, conidiation, or sterigmatocystin accumulation. During early growth, A. parasiticus and A. nidulans produced ethylene with approximately twofold higher quantities measured in continuous light than in the dark. 1-Methylcyclopropene (an inhibitor of ethylene receptors in plants), light, CO2, temperature, and growth medium composition altered the effect of ethylene on A. nidulans and A. parasiticus. These observations are consistent with the existence of an ethylene sensor molecule that mediates the function of an ethylene-responsive signaling pathway(s) in Aspergillus.

Relationship between ethylene response manipulation and volatile production in Jonagold variety apples.

The ripening of Jonagored Jonagold apple fruit (Malus x domestica Borkh.) during development was manipulated with preharvest applications of ReTain or a combination of ReTain plus Ethrel. The fruits, harvested preclimacteric at approximately the same stages of maturity, were stored in refrigerated air (RA) for 45 days or in controlled atmosphere (CA) for 180 days at 0 degrees C. Volatile evolution, ethylene production, and respiration of stored fruit were studied during poststorage holding at 22 degrees C. ReTain reduced volatile production by 19%, but application of Ethrel to ReTain-treated fruit increased production to control levels. The inhibition of volatile production by ReTain appears to be independent of respiration but may be related to the ethylene-producing capacity of the fruit. Although ReTain reduced flavor-related volatile esters, it did not affect levels of the compound responsible for the typical spicy flavor in Jonagored Jonagold fruit, 4-methoxy-2-propenylbenzene. The CA-stored fruit had a much reduced production of volatile compounds compared to RA-stored fruit, with more discernible effects in ReTain-treated fruit. Ethrel application to ReTain-treated fruit improved the volatile production intermediate between the ReTain alone and control in CA-stored fruit. The data collectively suggest that ReTain may have some promise for better scheduling of harvest of apples with no appreciable loss in RA-stored fruit quality. Reduction in production of alpha-farnesene by ReTain may also reduce the potential for scald development in CA-stored fruit.

Effect of superficial scald suppression by diphenylamine application on volatile evolution by stored Cortland apple fruit.

Cortland apple fruit (Malus x domestica Borkh.) stored for 120-140 days in air at 0 degrees C were warmed to 22 degrees C and held for 8 days. A portion of the fruit was dipped in a solution of diphenylamine (DPA) at harvest to prevent scald development. Scald occurred only in those fruit not treated with DPA, and its development was accelerated after transfer to 22 degrees C. Ester production from apple fruit tended to increase from day 0 to day 6 of poststorage holding and declined thereafter in both treatments. However, ester production in scald-developing fruit was reduced by approximately 50%. The reduction in volatile production remained relatively constant during the rapid development of scald symptoms. Furthermore, the reduction in volatile production appeared to be independent of respiration and ethylene production. Production of esters derived from hexanol was most reduced in fruit developing scald, with hexyl 2-methylbutanoate production being reduced approximately 15-fold. Interestingly, the production of methyl butanoate was detected only in scalding fruit. alpha-Farnesene production in fruit developing scald was reduced 43% compared with DPA-treated fruit. In contrast, the primary volatile oxidation product of alpha-farnesene, 6-methyl-5-hepten-2-one (MHO), was present only in fruit developing scald. The data suggest that inhibition of ester production may occur as a result of the physiological changes associated with susceptibility to, rather than expression of, scald symptoms.

Idiopathic ulcerative proctitis may be the initial manifestation of Crohn's disease.

Of 213 patients with proctitis of all etiologies seen between 1977 and 1987, we studied an original cohort of 96 patients with idiopathic ulcerative proctitis (mean follow-up of 62 months). The diagnosis was made according to strict inclusion criteria, and all cases with any initial feature suggestive of Crohn's disease were excluded. Follow-up showed that 13 patients (13.6%) eventually progressed to Crohn's disease, usually within the first 3 years of the initial diagnosis of idiopathic ulcerative proctitis. Their clinical, endoscopic and histological features at initial presentation were indistinguishable from those in whom the diagnosis remained that of idiopathic ulcerative proctitis. In nine of 13 of these patients (70.0%), the clinical course was characterized by a more protracted course and a poorer response to standard treatment. Such features were not found in those in whom the diagnosis of idiopathic ulcerative proctitis was maintained. Clinicians should be aware that Crohn's disease may present initially as apparent idiopathic ulcerative proctitis.

Banana ripening: implications of changes in glycolytic intermediate concentrations, glycolytic and gluconeogenic carbon flux, and fructose 2,6-bisphosphate concentration.

In ripening banana (Musa sp. [AAA group, Cavendish subgroup] cv Valery) fruit, the concentration of glycolytic intermediates increased in response to the rapid conversion of starch to sugars and CO(2). Glucose 6-phosphate (G-6-P), fructose 6-phosphate (Fru 6-P), and pyruvate (Pyr) levels changed in synchrony, increasing to a maximum one day past the peak in ethylene synthesis and declining rapidly thereafter. Fructose 1,6-bisphosphate (Fru 1,6-P(2)) and phosphoenolpyruvate (PEP) levels underwent changes dissimilar to those of G 6-P, Fru 6-P, and Pyr, indicating that carbon was regulated at the PEP/Pyr and Fru 6-P/Fru 1,6-P(2) interconversion sites. During the climacteric respiratory rise, gluconeogenic carbon flux increased 50- to 100-fold while glycolytic carbon flux increased only 4- to 5-fold. After the climacteric peak in CO(2) production, gluconeogenic carbon flux dropped dramatically while glycolytic carbon flux remained elevated. The steady-state fructose 2,6-bisphosphate (Fru 2,6-P(2)) concentration decreased to (1/2) that of preclimacteric fruit during the period coinciding with the rapid increase in gluconeogenesis. Fru 2,6-P(2) concentration increased thereafter as glycolytic carbon flux increased relative to gluconeogenic carbon flux. It appears likely that the initial increase in respiration in ripening banana fruit is due to the rapid influx of carbon into the cytosol as starch is degraded. As starch reserves are depleted and the levels of intermediates decline, the continued enhancement of respiration may, in part, be maintained by an increased steady-state Fru 2,6-P(2) concentration acting to promote glycolytic carbon flux at the step responsible for the interconversion of Fru 6-P and Fru 1,6-P(2).

Banana Ripening: Implications of Changes in Internal Ethylene and CO(2) Concentrations, Pulp Fructose 2,6-Bisphosphate Concentration, and Activity of Some Glycolytic Enzymes.

In ripening banana (Musa acuminata L. [AAA group, Cavandish subgroup] cv. Valery) fruit, the steady state concentration of the glycolytic regulator fructose 2,6-bisphosphate (Fru 2,6-P(2)) underwent a transient increase 2 to 3 hours before the respiratory rise, but coincident with the increase in ethylene synthesis. Fru 2,6-P(2) concentration subsequently decreased, but increased again approximately one day after initiation of the respiratory climacteric. This second rise in Fru 2,6-P(2) continued as ripening proceeded, reaching approximately five times preclimacteric concentration. Pyrophosphate-dependent phosphofructokinase glycolytic activity exhibited a transitory rise during the early stages of the respiratory climacteric, then declined slightly with further ripening. Cytosolic fructose 1,6-bisphosphatase activity did not change appreciably during ripening. The activity of ATP-dependent phosphofructokinase increased approximately 1.6-fold concurrent with the respiratory rise. A balance in the simultaneous glycolytic and gluconeogenic carbon flow in ripening banana fruit appears to be maintained through changes in substrate levels, relative activities of glycolytic enzymes and steady state levels of Fru 2,6-P(2).

Meningo-ependymitis in Whipple's disease.

Six years after apparent complete recovery from intestinal Whipple's disease, a 56 year old man developed insidious progressive somnolence and gait ataxia. Studies showed hydrocephalus with obstruction of the aqueduct and CSF leukocytosis and elevated protein. Arachnoid biopsy during craniotomy revealed chronic inflammatory infiltration with PAS-positive macrophages. The patient died 5 years later despite two courses of antibiotic therapy. This is the first report of histologically confirmed cerebral Whipple's disease during life. Whipple's disease is a systemic infectious disorder. Cerebral involvement even in neurologically asymptomatic patients should be sought with periodic CSF cytologic studies and a search for hydrocephalus. The possibility of cerebral Whipple's disease should be considered in the presence of unexplained hydrocephalus and/or chronic inflammatory changes in the spinal fluid, especially in those with past or active intestinal disease.