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Diagnóstico y tratamiento de las exacerbaciones

T RATAMIENTO INTEGRAL DE LA EEPOC

(molar %)

40

40

20

30

20

10

- 1.0

40

o.o

/r---,r---,?---,r---�7

6.0

7. 0

8.0

A

(pH)

1.0

FIGURE 4 . 14 Predi cted 7-Ketodeoxycholate Yield

74

- 1.0,

N2-co2

N2

(Gas)

The model from which this figure is drawn i s pre s ented in Appendix 2 , Tab le A2 . 1

L. M.

C.

t

8. 6

8.2

1.0

o.o

/�---,�---�,�---,r---�7

6.0

--!$

8.0

A

(pH)

(Gas)

1 . 0

FIGURE 4 . 15 Predicted Ce l l Yield ( as L . M . C . , of maximum observed number o f ce lls mi llili tre o f culture )

7 5

The model from which this figure i s drawn i s presented in Appendix 2 , Table A2 . 5 .

TABLE 4 . 4

c DC L . M. C .

Matrix o f Correlation Coe f fi cients for the P airs of

Vari ab les in 1

7KD c D

- 0 . 876 * * *

0 . 266 - 0 . 6 9 8 *

0 . 772 * * * - 0 . 8 84 * * * 0 . 6 20 * * * * - s tatistically s igni fi cant at the 1% leve l

* - s tatistically s i gni fi cant at the 5 % level

4 . 5 DI SCUSS ION OF EXPERIMENT l

The expe rimental de sign approach adopted i s empi ri cal , and as s uch i t is s ui ted to the proj e ct aim. Th is me ans that any mechanistic dis cus s ion indulged in i s speculative , but may ass is t in the i dentifi­ cation of areas worthy o f further res earch .

The regress i on mode l s con fi rm what i s intui tive ly obvious from the raw data ; the pre fe rred conditions for dehydroxylation lie very c lose to pH 7 with an atmosphe re of N 2 - co2 , 9 : 1 , whe re as for de­ hydrogenation , an atmo sphe re o f nitrogen , close to pH 7 is optimum.

7 6

I f an experimental des i gn with only two levels ( i . e . a 2 2 factorial ) had been s e lecte d , the dehydroxylation optimum might not have been dete cte d , since the s igni fi cant negative main e ffe cts for both vari ables would le ad to the con c lus i on that greatest deoxycholate yields lay in the dire ction o f low pH and hydrogen gas . The three­

leve l design adopted allowed the e l ucidation of much curvature in b oth variables , reflected in the s i z e of quadratic terms in all mode ls , and e vi denced by .the ability o f a logari thmic trans formation to des cribe deoxycholate yield. The h i gh pe ak for dehydroxylation at pH 7/Nz-COz i s mani fest mai n ly in inte ractions involving quadratic terms . Thi s p eak i s o f great infl uence i n the mode l , yet comprises only a s ingle data point . The regression techniques us ed tend to weight s uch large outlying points , and in this case the apparent statistical signi fi -

cance is enhanced by c lose agreement of the repl icates . Moreove r , this high value arose i n the very fi rs t run , whe re di ffe rent random in fluences may have been acting .

To check this value , and t o sub stantiate the predicted maximum deoxycholate yield (wh i ch corresponded clos e ly to run 1 condi tions ) , run 1 was repeated. The results are depi cted in Figure 4 . 16 . The total degree o f t ran s formation was as predic te d ( 50 . 6 % , p rediction 50 . 0 % ) and growth yield on ly a little high ( L . M. C . 8 . 80 , predi ction 8 . 60 ) . Howeve r , the yield of deoxycholate was lowe r than predicted

via

the logari thmi c t rans formation ( 10 . 9 % , predi ction 20 . 3 % ) and

7-ketodeoxycholate yield correspondingly higher ( 38 . 5 % , predicted 3 1 . 0 % ) . The diffe rence between the two pH 7/N 2 -c02 runs may reflect

" a change in the random in fluence s affecting any experiment . Thi s i s parti cularly like ly s ince the 1 1- run des ign was performed a s a s i ngle

randomi sed block ove r 3 months , and the repe at some weeks late r on its own . De spite the di ffe rence , the repeat run s till showed a highe r deoxychol ate yield than any of the othe r ten t rials . Thus the bas i c conclus ion still holds : dehydroxylation was gre atest at pH 7 unde r the N2 -C02 mixture .

Most lite rature reports concerning dehydroxylation as a function of pH have de alt with rates o f trans formation . The current work has

' been concerned with yie lds , which are more relevant to the e arly

\stages of proce ss deve lopment , and requi red fewer data points for adequate calculation . Only when deoxychol i c acid yields be come fav­ ourable will rates become of intere s t from a p rocess viewpoint . This di fference between yie ld optima and rate optima mus t be borne in mind when cons ide ring p revi ous ly reported pH " optima" .

7 7

The l iterature s hows some dis agreement concerning the dependence of dehydroxylation on pH . Dras ar and Hi l l ( 19 74 ) reviewed thei r own extensi ve faecal-isolate screening experiments , and simi lar expe riments of thei r col le ague s , all of whi ch emp loye d T . L . C . to detect 7a-dehyd­ roxylation by harvested and res uspended ce l l s of anae robically grown organisms . They conc luded that i f the pH o f the culture medium fe l l below p H 6 . 5 , very l i ttle dehydroxylation could b e detected. Midtvedt and Norman ( 1968) investigated growing c ultures of thei r lactobacillus-

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