Fecal Fraction Research Articles

Steroid ecretion in he neo-natal period: A comparative study of the excretion of steroids by human, ape and rhesus monkey infant

Analysis by gas chromatography-mass spectrometry of the steroid sulphates present in urine and faeces from a 3-week-old orang-utan indicated that, as in the human infant, 3β-hydroxy-5-ene steroids are the major urinary steroids. The production of these steroids in the neonatal period is carried out by the foetal zone of the adrenal gland which does not disappear in ape and human infants until several weeks after birth. The major steroids identified in urine were: 3β,16α-dihydroxy-5-androsten-17-one, 3β,17β-dihydroxy-5-androsten-16-one, 5-androstene-3β, 16α, 17β-triol, 5-androstene-3β, 16β, 17α-triol, 3β, 16α-dihydroxy-5-pregnen-20-one and 3β,21-dihydroxy-5-pregnen-20-one. A pregnenetriol, an androstenetetrol, an androstenetriolone, tetrahydrocortisone and cortolone were also detected. The major steroids identified in the steroid sulphate fraction of faeces were: 3β-hydroxy-5-androsten-17-one (DHA), 3β-hydroxy-5-pregnen-20-one (pregnenolone), 5-androstene-3β,16α,17β-triol, 5-androstene-3β, 16β, 17α-triol, 5-pregnene-3β,16α,20α-triol, 5-pregnene-3β,20α,21-triol and 24-hydroxy cholesterol. The total amounts of steroid sulphates excreted in urine and faeces were similar. The urinary steroids of newborn rhesus monkeys were all “saturated”, no 3β-hydroxy-5-ene-or 16-hydroxylated steroids were detected. The following were identified: androsterone, 5β-pregnane-3α,17α,20α-triol, 3α,21-dihydroxy-5β-pregnane-11,20-dione and tetrahydrocortisone. The lack of 3β-hydroxy-5-ene steroids from urine is due to the disappearance of the foetal zone prior to birth in this species.

Decrease of excretion of cholesterol during aging

Excretion of tritium was measured in feces after i.v. injection of 3H-cholesterol to young (2·5 months) and old (14 months) rats. Excretion of catabolic products of 3H-cholesterol was by about 70 per cent smaller in older animals which may partly explain their higher retention of 3H-cholesterol in the blood. Excretion of 3H-cholesterol in the steroid fraction of feces was decreased more than excretion of tritium in the rest of feces which is mainly bile acid fraction. After ligature of the common bile duct which excludes bile secretion the decrease of excretion of 3H-cholesterol in older animals remains, showing that also intestinal secretion of cholesterol is smaller in older rats. Possible effect of smaller spontaneous food intake of older animals on excretion of cholesterol was excluded by pair-feeding of the younger animals. A hypothesis is advanced that formation of bile acids from cholesterol is decreased in older animals. After parabiotic connection of young and old rats, excretion of 3H-cholesterol was increased in the old parabiotic partners and also turnover of 3H-cholesterol tissues was increased to the level observed in young animals. This points to a presence of humoral factors in the blood of young and old animals effecting metabolism of cholesterol.

Migratory behaviour of diapausing larvae of Trogoderma granarium (Coleoptera, Dermestidae) in relation to odours produced in their culture medium

The olfactory response of diapausing larvae of Trogoderma granarium to odours produced in their culture medium was investigated in order to clarify whether or not these odours influence the larval migratory behaviour. Tests were carried out with larval faeces and wheat, and extracts and fractions thereof, at 33°C and 40–45% r.h. The results show that diapausing larvae react differently to wheat or faeces odours according to the faecal content of the medium in which they are feeding. The faecal fatty acids were found very attractive, but they were less attractive than the intact faecal extract; the unsaponifiable faecal fraction was relatively ineffective. The migratory behaviour of diapausing larvae is controlled at least in part by odours of faeces or wheat prevailing in the environment.

Sulphur metabolism and excretion studies in ruminants. IV. Cystine and sulphate effects upon the flow of sulphur from the rumen and upon sulphur excretion by sheep

In a 4 x 4 Latin square design experiment, sheep were fed on a basal ration which supplied 0.61 g sulphur per day, or the basal ration supplemented with 1.4 g inorganic sulphate sulphur, or 1.4 g cystine sulphur, or 1.4 g sulphate sulphur plus 1.4 g cystine sulphur per day. All forms of sulphur supplements increased the daily flow of protein sulphur from the rumen to the omasum (P < 0.05) above basal. Only the cystine treatments increased the flow of sulphide (P < 0.005), ester sulphate (P < 0.005), soluble organic sulphur (P < 0.05), and cystine (P < 0.05), and increased the ruminal sulphide concentrations above basal (P < 0.005). From 7.0 to 12.6% of the added cystine was not degraded in the rumen. In three animals less than 3 % of the added sulphate, alone or in combination with cystine, passed unchanged to the omasum. The fourth animal (67) differed from the others (P < 0.05): the added sulphate was poorly reduced in the rumen, and 62% reached the omasum unchanged. The sulphur composition of the digesta collected from the omasum was: protein sulphur, 55-70%; soluble organic sulphur, 13-22%; inorganic sulphate sulphur (excluding sheep 67), 2.2-2.6 %; ester sulphate sulphur, 1.8-3.4 %; and sulphide, 0.4-28%. Excluding sheep 67, 36% of the dietary sulphur was absorbed from the rumen when the sulphur was added as sodium sulphate or as cystine, and 50 % when added in both forms together. There was a net addition of 0.45 g sulphur per day to the rumen on the basal diet. The influence of the amount and the form of the dietary sulphur on the excretion of faecal and urinary sulphur fractions is discussed.

The function of fatty acids in the diapause of the khapra beetle Trogoderma granarium everts

1. 1. Dietary fatty acid fraction of larval faeces of Trogoderma granarium, containing palmitic, stearic, oleic and linoleic acids, induces and maintains the diapause in the larvae of the same species; linoleic acid is highly efficient either by feeding or by injection. 2. 2. A comparative examination of the fatty acid composition of diapause and nondiapause larvae of T. granarium showed that the diapause insects have an increased concentration of the bound myristoleic, palmitoleic and linoleic acids accompanied by a decrease in palmitic acid content. 3. 3. Mass spectra, infra-red and R f values for fatty acid esters of larval faeces are given.

Steroids in germfree and conventional rats. Steroids in the mono- and disulphate fractions of faeces from female rats.

Gas‐liquid chromatography, radio‐gas chromatography and gas chromatography‐mass spectrometry were used to identify steroid metabolites in the monosulphate and disulphate fractions of faeces from germfree female rats and in the monosulphate fraction of faeces from conventional female rats after intraperitoneal administration of [4‐14C]pregnenolone and [4‐14C]‐corticosterone. The germfree rats excreted the following steroids: monosulphate fraction: 3β, 17α‐dihydroxy‐5α‐pregnan‐20‐one, 5α‐pregnane‐3α,16α,20α‐triol, 2α,3β,16α‐trihydroxy‐5α‐pregnan‐20‐one and 3α,15β,16α‐trihydroxy‐5α‐pregnan‐20‐one; disulphate fraction: 3α‐ and 3β,11β,21‐trihydroxy‐5α‐pregnan‐20‐one. The following steroids were identified in faeces from conventional rats: oestriol, 3α,15α(and 15β)‐dihydroxy‐5α,17α(and 17β)‐pregnan‐20‐one,3α,19‐dihydroxy‐5α,17α‐pregnan‐20‐one, 5α‐pregnane‐3α,16α,20‐triol, 3β,11α‐dihydroxy‐5α‐pregnan‐20‐one. 2α, 3α‐dihydroxy‐5α,17α‐pregnan‐20‐one (tentatively identified), and 3α,17α‐dihydroxy‐5α‐pregnan‐20‐one. The different steroid patterns in faeces from germfree and conventional rats are explained by the metabolic reactions carried out by the intestinal flora. Radio‐gas chromatography showed that 3α‐ and 3β,11β,21‐trihydroxy‐5α‐pregnan‐20‐one and 3β,11α‐dihydroxy‐5α‐pregnan‐20‐one were metabolites of corticosterone. The formation of 3β,11α‐dihydroxy‐5α‐pregnan‐20‐one and of the 17α‐pregnane derivatives by intestinal microorganisms is discussed.

Identification of C19O2 and C21O2 steroids in the mono- and disulphate fractions of human faeces.

Steroid monosulphate and disulphate fractions were obtained from human faeces by chromatography on Sephadex LH‐20 and solvolysed. The steroids liberated were fractionated on silicic acid. Fractions containing C19O2 and C21O2 steroids were studied by thin‐layer chromatography, gas‐liquid chromatography and gas chromatography‐mass spectrometry.Dehydroepiandrosterone, 5α‐androstane‐3α,17α‐diol, 5α‐androstane‐3α,17β‐diol, 5α‐androstane‐3β,17α‐diol, 5α‐androstane‐3β,17β‐diol, 5β‐androstane‐3α,17β‐diol, 5‐androstene‐3α,17β‐diol, 5‐androstene‐3β,17α‐diol, 5‐androstene‐3β,17β‐diol, 5α‐pregnane‐3α,20α‐diol, 5α‐pregnane‐3β,20α‐diol, 5β‐pregnane‐3α,20α‐diol, 5β‐pregnane‐3β,20α‐diol,pregnenolone, 5‐pregnene‐3α,20α‐diol and 5‐pregnene‐3β,20α‐diol were found in the monosulphate fraction and 5‐androstene‐3β,17α‐diol as well as 5‐androstene‐3β,17β‐diol in the disulphate fraction of human faeces.Several steroids identified earlier in the disulphate fraction of human bile were found in the monosulphate fraction of human faeces, indicating partial hydrolysis of diconjugates in the intestine. The androstanediols and pregnanediols found in the faeces were primarily of a 3β‐hydroxy‐5α‐ or 3β‐hydroxy‐5β‐structure, whereas those in the bile were of a 3α‐hydroxy‐5α‐ or 3α‐hydroxy‐5β‐structure. This suggests conversion of a 3α‐hydroxyl to a 3β‐hydroxyl in the intestine in vivo.

Steroids in newborns and infants. C19 and C21 steroids in faeces from infants.

The steroids in faeces from infants less than one year of age have been studied. Two fractions, monosulphates and “diconjugates”, were separated by chromatography on Sephadex LH‐20. The steroids were liberated by solvolysis and were studied by thin‐layer chromatography and gas‐liquid chromatography‐mass spectrometry. Eighteen steroids, most of them having a 3β,16‐dihydroxy‐Δ5 structure, were identified in the monosulphate fraction of pooled faeces from infants 1–4 months of age. Of these, androst‐5‐ene‐3β,16β,17α(and)17β‐triols, 3β‐hydroxypregna‐5,16‐dien‐20‐one, 3β,16β‐dihydroxypregn‐5‐en‐20‐one, 3α,16α‐dihydroxy‐5β‐pregnan‐20‐one, pregn‐5‐ene‐3β,16α,20α‐triol, pregn‐5‐ene‐3β,20α,21‐triol and pregn‐5‐ene‐3β,16α,20α,21‐tetrol have not been reported as occurring in urine from infants. In addition, eight steroids were found, the stereochemistry of which was not confirmed. Four of the 16‐hydroxylated steroids were also present in the “Diconjugate” fraction.In contrast to faeces from the younger infants, pooled faeces from infants 6–12 months of age contained only small amounts of steroids, mainly pregn‐5‐ene‐3β,20α‐diol and pregn‐5‐ene‐3β,17α,20α‐triol. The results indicate that significant amounts of steroids are excreted in faeces during the first few months of life, and that metabolism of these steroids by the intestinal flora is of minor importance in this period.

Identification of 22-, 24- and 26-hydroxycholesterol in the steroid sulphage fraction of faeces from infants.

A fraction of solvolyzable sterol conjugates with the chromatographic properties of steryl sulphates was isolated from pooled faeces collected from infants 1–4 and 6–12 months old. The major sterols in this fraction were analyzed by gas chromatography‐mass spectrometry. The pool from the younger infants was shown to contain cholesterol and 22α‐, 24χ‐, and 26‐hydroxycholesterol and the pool from the older infants contained cholesterol, campesterol, β‐sitosterol, and 22α‐, and 24χ‐hydroxycholesterol. The concentrations of cholesteryl and 24‐hydroxy‐cholesteryl sulphates were higher in faeces from the older than from the younger subjects.

The quantification of cholesterol excretion and degradation in the isotopic steady state in the rat: the influence of dietary cholesterol

A means of quantifying the two major pathways of cholesterol elimination from the body, bile acid production and neutral sterol excretion, has been devised for use in the intact rat. Utilizing an isotopic for blood cholesterol (which has been attained by the subcutaneous implantation of capsules containing cholesterol-4-C14) and the measurement of C14 appearance in the bile acid and sterol fractions of feces, it is possible to measure the amounts of these two excretion products. With this method the increase in bile acid formation following cholesterol feeding has been demonstrated directly; this increase has been shown to be sufficient in magnitude to account for the entire positive balance for neutral sterol which occurs in the cholesterol-fed animal after equilibrium has been attained. Finally, utilizing a double isotopic steady state in animals fed cholesterol-7α-H3 and implanted with cholesterol-4-C14, it has been possible to demonstrate that the major portion of fecal cholesterol of endogenous origin is derived from or is in equilibrium with that of the blood. In these animals the measurement of dietary cholesterol absorption has also been possible.

METABOLISM AND EXCRETION AS FACTORS INFLUENCING SERUM CHOLESTEROL LEVEL IN TWO LINES OF CHICKENS.

SummaryTwo lines of White Leghorn chickens differing in serum cholesterol level were studied. The line difference in cholesterol was unchanged in serum from post-hepatic blood as compared to pre-hepatic blood. No line differences were observed in rate of in vitro cholesterogenesis from acetate-1-C14 incubated with blood or liver slices. Following administration of cholesterol-4-C14, differences were not detectable between lines in labeled nonsaponifiable substances in liver, gut, skin, adrenal, or carcass, or in the labeled saponifiable fraction of liver or feces. Fecal excretion of labeled nonsaponifiable substances, however, was significantly higher in the low line, suggesting genetic regulation of cholesterol homeostasis through excretory-pathways.Grateful acknowledgement is made to Dr. Sitarama Lakshmanan for consultation on radioisotope techniques and for the use of his laboratory.

On the Composition of the Bile Acid Fraction of Rabbit Feces and the Isolation of a New Bile Acid: 3α,12α-Dihydroxy-5α-cholanic Acid: BILE ACIDS AND STEROIDS 136

On the Composition of the Bile Acid Fraction of Rabbit Feces and the Isolation of a New Bile Acid: 3α,12α-Dihydroxy-5α-cholanic Acid: BILE ACIDS AND STEROIDS 136

URINARY AND FECAL CONVERSION PRODUCTS OF 16-OXOESTRADIOL-17β-16-C14IN THE DOMESTIC FOWL

16-Oxoestradiol-17β-16-C14was injected intravenously into non-laying hens and the urine and feces for succeeding days were collected separately. Estrogen extracts were prepared using enzymic hydrolysis. Analyses of the urinary phenolic fractions showed the presence of two major radioactive products and one minor radioactive product. The major products were identified as 16-oxoestradiol-17β and 16-epiestriol. The minor product was not identified, but it contained little if any radioactive estriol. The fecal phenolic fraction contained a radioactive material identified as 16-epiestriol. A summary of recoveries and distributions of the radioactivity is presented along with criteria for the radiochemical purities of the isolated steroids.

Weight gain and faecal nitrogen excretion in grazing British and Zebu crossbred steers

The excretion of faecal nitrogen by 16 grazing steers, eight Hereford x Shorthorn and eight Zebu crossbreds, was determined at intervals over a period of 11 months. It was found that the non-dialysable faecal nitrogen fraction was the one most strongly correlated with gain. Statistically this fraction accounted for 68% of the variation in average gain of the two breed groups between measurement intervals, and for 75% of the variation in weight gain between individual steers during the summer gain period. The data gave no indication of inherent breed differences in intake, as judged by non-dialysable nitrogen excretion.

Influence of dietary cholesterol on excretion of cholesterol-4-C14 by the rat.

The influence of dietry cholesterol on the excretion of cholesterol-4-C14 has been studied in rats pair fed isocaloric quantities of purified diets supplemented with varying amounts of cholesterol and either oleic or linoleic acid. Of the C14 excreted over an 8-day period, slightly more than half was present in the neutral sterol fraction of feces from rats fed sterol-free diets. The addition of dietary cholesterol, however, under circumstances where significant cholesterol absorption was promoted by the addition of fatty acids to the diets, resulted in a reversal of this ratio so that the bile acid fraction accounted for the major portion of excreted cholesterol-C14. Cholesterol balance data revealed that the degradation rate of cholesterol in these animals rose to about 17–20 mg/day. These data are interpreted as demonstrating that the amount of absorbed dietary cholesterol plays a crucial role in determining both the relative and absolute importance of the two major routes of cholesterol excretion.

Effect of soybean sterols on liver deposition of cholesterol-C14.

1. The effect of soybean sterols A on liver deposition and fecal excretion of cholesterol-C14 following a single feeding of I the labeled material has been studied using cholesterol-fed rats. Average recoveries of J labeled cholesterol in liver (measured as radioactivity recovered from liver non-saponifiable fraction) were 6.61% in a cholesterol-fed group, 1.80% in a cholesterol-fed group receiving one g of soy sterol immediately after cholesterol-C14 administration and 0.47% in a group fed both cholesterol and soybean sterol for 120 days. 3. Average recoveries of radioactivity in the non-saponifiable fraction of feces were 20.14%, 25.13%, and 27.76%, respectively, for the 3 groups described above.