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HPF = high-power field.
Figure 3.
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Figure 3.

Inhibition of chemotaxis by various preparations and the effect of clarification. Three separate preparations of chicken soup were tested either in the completed stage (solid bars) or after clarification by centrifugation (15 min, 12,000g, stipple bars). All samples were diluted 1:40 into HBSS and were added together with the neutrophils. ZAS, diluted 1:4, was used as the chemoattractant. HBSS and ZAS alone are shown as negative and positive controls, respectively. Vertical axis: migrated neutrophils. Horizontal axis: conditions. * = p < 0.05 compared to ZAS control. See Figure 2 for other abbreviations.

In an attempt to partially determine which components of chicken soup had activity, two experimental approaches were undertaken. First, samples of chicken soup were harvested at various times during the preparation. As the soup preparation was exceedingly inhomogeneous, samples were taken from various parts of the pot at various times. Samples containing the initial stages of the soup with early chicken broth alone were not active (Fig 4 ). All samples harvested after the addition of the first group of vegetables had inhibitory activity. In the final stages of the preparation, slightly less inhibition of chemotaxis was observed. Second, an analysis of inspanidual soup components was performed by boiling inspanidual components. All ingredients were found to be inhibitory, including the boiled extract of chicken alone (Fig 5 , top). The effect on the inhibition of the whole soup was not due to effects on neutrophil viability. The viability of neutrophils exposed to each of the complete chicken soup preparations was always > 95%. The viability of the neutrophils incubated in the boiled chicken stock was 98%. The isolated vegetable components, interestingly, demonstrated a slight, but statistically significant, loss of neutrophil viability, as assessed by trypan blue exclusion (Fig 5, bottom).
Figure 4.
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Figure 4.

Acquisition of chemotactic inhibitory activity during preparation of the soup. Samples were collected at various stages during the preparation of chicken soup. Each aliquot then was diluted 1:100 into HBSS and was added together with neutrophils in the top portion of the chemotaxis chamber. Chemotaxis then was measured using ZAS diluted 1:4 as the chemoattractant. The major stages of the soup preparation are indicated. See Figure 2 for other abbreviations.
Figure 5.


Figure 5.

The effect of specific vegetable ingredients on the inhibition of chemotaxis and neutrophil viability. Top: the component ingredients of chicken soup were boiled in water after which they were sonicated, diluted 1:100 in HBSS, and added to neutrophils in the top portion of a chemotaxis chamber. ZAS diluted 1:4 was used as the chemoattractant. Bottom: viability was assessed by trypan blue dye exclusion after incubating the neutrophils in HBSS with the diluted soup components for 30 min. See Figure 2 for other abbreviations.

In a modest attempt to determine whether commercially available preparations of chicken soup also inhibited neutrophil chemotaxis, 13 different soups were purchased at a local supermarket and were tested against Grandma’s soup (Fig 6). Many of the soups inhibited neutrophil chemotaxis. Five inhibited more potently (at an identical dilution) than did Grandma’s traditional soup. Two soups were without activity, and one slightly augmented chemotaxis. Omaha tap water had no activity.


The effect of various commercial soups on neutrophil chemotaxis. A number of preparations of soup were purchased at a local supermarket and were prepared according to the instructions on the label. All samples then were diluted 1:40 in HBSS added to neutrophils, and chemotaxis to ZAS (diluted 1:4) was tested. See Figure 2 for other abbreviations.
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Discussion

The current study demonstrates that chicken soup inhibits neutrophil migration to standard stimuli as assessed by the modified Boyden blindwell chamber method. The effect appears to be due to an effect on the neutrophils rather than on the chemoattractant, as addition of the soup directly to the neutrophils appears to be most effective. The inhibitory effect was observed clearly at concentrations without cytotoxicity, as determined by trypan blue dye exclusion. Finally, a variety of soup preparations was evaluated and found to be variably, but generally, able to inhibit neutrophil chemotaxis. The current study, therefore, presents evidence that chicken soup might have an anti-inflammatory activity, namely, the inhibition of neutrophil migration.

The identity of the active ingredient or ingredients present in the soup remains unknown. The vegetables that are used to prepare the soup, however, are known to contain a large number of chemical species, many of which have medicinal activities.252627 A number of fats and substances with antioxidant activity are also likely to be present. Extracts of each vegetable, as well as of the chicken, all were able to inhibit neutrophil chemotaxis, suggesting that many inhibitory substances may be present. Interestingly, the vegetable extracts also demonstrated some neutrophil cytotoxicity that was not observed either in the completed soup or in the chicken extract. No attempt was made to control for concentration of various extracted components, and the toxicity could be due to a concentration-dependent effect

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