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Biomedical subjects

I R Fleet

Publications and source records attributed to I R Fleet.

At least 37 records · Page 2Linked to original sources

Suppression by perchlorate of technetium-99m and iodine-123 secretion in milk of lactating goats.

Lactating goats were infused with either technetium-99m (99mTc) or iodine-123 (123I) together with chlorine-36 (36Cl) through an indwelling catheter previously placed in an external pudic mammary artery. The radioisotope infusions were repeated together with 100 mg of sodium perchlorate. There was a rapid transfer of 99mTc and 123I into milk, reaching a peak concentration 30 min after a 15-min infusion. The fractional secretion of 99mTc and 123I in milk was reduced by 70%-80% and 60%-66%, respectively, by perchlorate. The fractional secretion of 36Cl was not affected by perchlorate, and the shape of the 36Cl secretion curve differed from those of 99mTc and 123I, which were similar. It is probable, therefore, that the latter nuclides were secreted by a transport route different from that of chloride. Available data describing the secretion of 99mTc in human milk after pertechnetate administration was reviewed, and it was concluded that perchlorate pretreatment significantly reduced the secretion of 99mTc in human breast milk.

Animals↗

Residual milk in Friesland sheep and the galactopoietic effect associated with oxytocin treatment.

Mature lactating Friesland ewes had a mean lactation yield of 293 +/- 26 kg during a lactation period of 35 +/- 2 weeks giving an average daily milk yield of 1.2 kg/d. Ewes were injected intravenously after normal milking with either saline (sham) or oxytocin and then remilked to determine the volume of residual (alveolar) milk. After a long milking interval of 16 h oxytocin treatment gave a significantly greater total daily milk yield than the sham treatment (oxytocin minus saline, morning milking, 0.199 +/- 0.038 kg, mean +/- s.e.m., P less than 0.01) Oxytocin had a small significant reverse effect after a short milking interval of 8 h (afternoon milking, -0.065 +/- 0.022 kg, P less than 0.05). The average increase in total daily yield over four stages of lactation was 0.133 +/- 0.029 kg (P less than 0.01), or 11% of the average daily milk yield. The galactopoietic effect of oxytocin was associated with the efficient removal of residual (alveolar) milk. Residual milk accounted for 7.4 and 27.2% of the total daily milk yield in ewes treated with saline or oxytocin respectively. Residual milk expressed as a proportion of daily total milk yield remained steady in ewes studied between April and July, but declined in September when yields were less than 1 kg/d.

Animals↗

Kinetics of transfer of 125I-labelled epidermal growth factor from blood into mammary secretions of goats.

125I-Labelled mouse epidermal growth factor (125I-EGF) was transferred intact and undegraded from circulating blood into milk in conscious lactating goats. Greater than 90% of the total radioactivity present in milk from the infused gland was in the aqueous phase and more than 72% was acid-precipitable. This radiolabelled material co-eluted with authentic EGF through gel filtration and was immunoprecipitable by a specific rabbit anti-mouse EGF immunoglobulin. Mammary uptake of 125I-EGF infused into mammary arterial blood (close-arterial infusion) for 1 h varied from 20 to 83% at different stages of the reproductive cycle. Only 0.5-2.9% of the infused 125I-EGF was transferred into milk during the first 3 h after the start of the infusion, which represents 0.7-6.3% of mammary uptake of EGF. The kinetics of transfer of 125I-EGF were followed in two lactating goats. Radioactivity reached peak levels in milk about 120 min after the start of a 1 h close-arterial infusion into the mammary gland, with an initial lag of about 30 min when little transfer occurred. Transfer was slower in two non-lactating goats with maximal levels of activity in milk being reached after about 180 min. The results are consistent with a transcellular transfer, whereby the factor is bound to receptors on the baso-lateral membrane, internalized by epithelial cells and subsequently secreted across the apical membrane into the alveolar lumen. The low level of degraded labelled EGF in milk (and mammary vein blood) suggests a modification of the normal pathway of EGF degradation such that the delivery of internalized factor to lysosomes is avoided.

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Mechanisms of transfer of steroid hormones and growth factors into milk.

In this paper we examine the ability of the mammary gland to remove from circulating blood three compounds which differ in their physico-chemical and structural properties. Mammary extraction of progesterone, oestrone sulphate and epidermal growth factor (EGF) is similar at peak lactation in goats, but the proportion of labelled infusate that is transferred into milk is greater for oestrone sulphate and EGF than progesterone which is rapidly metabolised by mammary tissue. The kinetics of transfer of progesterone, oestrone sulphate and EGF from blood into milk show that transcellular processes are involved, and on the basis of earlier hypotheses and new information reported here the results indicate the probable importance of simple and facilitated diffusion pathways for progesterone and oestrone sulphate, and secretory mechanisms for oestrone sulphate and EGF. Although evidence is lacking for a direct effect of hormones in milk on mammary function, their concentration in milk may reflect changes in local regulation of mammary secretion. Considerable practical value is attached to the immunodiagnostic use of milk hormone concentrations to determine ovarian and placental endocrine activity during pregnancy in domestic ruminants.

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Mammary uptake of amino acids and glucose throughout lactation in Friesland sheep.

Changes in mammary blood flow, arterial and venous plasma concentrations of glucose and individual amino acids, udder volume, and milk yield and composition were measured at intervals throughout lactation in four Friesland ewes. Milk yields peaked 50-80 d post partum and declined by 40% within 3 months. Neither mammary blood flow (43.3 +/- 5.8 (s.e.m.) ml/100 cm3 X min) nor udder volume changed significantly throughout the period of study, but for three ewes the 'mammary blood flow: milk yield' ratio increased from 300 (peak yield) to 570 (late lactation). Mammary glucose uptake remained essentially constant throughout lactation despite a 50% decline in lactose output. Arterial concentrations of glucose were much lower at peak yield than in late lactation. Mammary amino acid uptake conformed quite closely to 'essential' and 'non essential' categories previously defined for goats and cows, the degree of balance with output in milk protein being similar at all stages of lactation. For several amino acids arterial concentrations and arteriovenous differences were significantly positively correlated: the changes in arterial concentrations with lactation stage were also correlated for some amino acids. Apart from the intrinsic value of such studies on a breed of ewe increasingly used for dairy purposes, the Friesland ewe appears well suited for use in quantitative metabolic studies on lactation.

Amino Acids↗

Prostaglandin F-2 alpha is transferred from the uterus to the ovary in the sheep by lymphatic and blood vascular pathways.

[3H]Prostaglandin F-2 alpha (PGF-2 alpha) was infused into a uterine lymphatic vessel or a uterine vein for up to 1 h, or injected into the uterine lumen of anaesthetized non-pregnant sheep 7-15 days after oestrus. After an intraluminal injection, labelled PGF-2 alpha was recovered in uterine lymph and peak radioactivity was reached 50 min after injection. [3H]PGF-2 alpha infused at a constant rate into a uterine lymphatic vessel resulted in a maximum concentration of radioactivity in plasma which was 5.6- and 1.7-fold higher in the adjacent utero-ovarian and ovarian vein, respectively, than in carotid arterial plasma. Estimation of the amount of infusate transferred from a lymphatic into ovarian venous blood gave a value (0.4%) similar to that for transfer from a uterine vein (0.3%). Evidence for local transfer was substantiated by the presence of significantly higher concentrations of 3H-labelled compounds in the ovary and corpus luteum adjacent to the site of intra-lymphatic infusion compared with those in the opposite organs. The concentrations in the adjacent ovary and corpus luteum were significantly greater when an intra-lymphatic rather than intra-uterine vein infusion was adopted. The results show that [3H]PGF-2 alpha is transferred locally from uterine lymphatic vessels into the adjacent ovary, corpus luteum and ovarian vein.

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Transport of oestrone sulphate by the mammary gland in the goat.

During pregnancy in goats the concentration of endogenous oestrone sulphate in milk increased more than twofold, and that in arterial and mammary venous plasma 10- and 20-fold respectively. The concentration in milk was higher than that in arterial plasma, particularly in lactating goats during mid-gestation. This was partly related to mammary production of oestrone sulphate (or of a closely related steroid which cross-reacted in the radioimmunoassay) since in tracer infusion studies the specific activity of oestrone sulphate in milk was significantly lower than that in arterial or mammary venous plasma. It was also related to the existence of a mechanism within the gland which concentrates oestrone sulphate in milk since when infused close-arterially into the mammary gland of a non-pregnant goat with undetectable levels of the endogenous compound in the circulation, a concentration ratio of 7.4:1.0 was reached for oestrone sulphate in milk:arterial plasma. Tracer kinetic studies showed that mammary extraction of [3H]oestrone sulphate was variable (up to 41.3 +/- 30.6%, mean +/- S.E.M.). During intravenous or close-arterial infusion, radioactivity in arterial and mammary venous plasma at steady state was mainly in the form of [3H]oestrone sulphate (range, 64 +/- 10.6 to 80.2 +/- 5.9% of total radioactivity in plasma). The remainder was in the form of compounds chromatographically similar to oestradiol-17 beta-3-monosulphate, oestradiol-17 alpha-3-monosulphate and unconjugated oestrogens. The distribution of radioactivity between these different steriods was similar in arterial mammary venous plasma indicating a low level of selective mammary metabolism or extraction. The amount of labelled oestrone sulphate transferred into milk was low, and it was significantly less in pregnant (range, 0.11 +/- 0.07 to 0.27 +/- 0.16% of total infusate) than in non-pregnant animals (3.23 +/- 0.50%). Studies of the rate of transfer of [3H]oestrone sulphate from blood to milk indicated the presence of a transcellular route with peak activity in milk occurring about 110 min after the start of the infusion.

Animals↗

Oestrogen metabolism in the endometrium, corpus luteum and ovarian residual tissue of the rabbit.

Reproductive tissues (uterine endometrium, corpus luteum and ovarian residual tissues) from pregnant and pseudopregnant rabbits were incubated with equimolar concentrations of [3H]oestrone and [3H]oestrone sulphate (0.375 pmol) to monitor the changes in oestrogen metabolism during the early stages of pregnancy (days 0, and 3-8 post coitum) and to investigate the embyonic effect upon maternal oestrogen metabolism. Oestradiol-17 beta was the major metabolite formed from oestrone and sulphoconjugation occurred in all tissues studied. Oestrone sulphate was converted primarily to oestradiol-17 beta-3-monosulphate. Endometrial 17 beta-oxidoreductase significantly decreased and sulphotransferase increased in activity during the preimplantation period, but no differences were noted between gravid and non-gravid horns in unilaterally pregnant animals, nor between pregnant or pseudopregnant animals. Significant decreases occurred in 17 beta-oxidoreductase and sulphotransferase activity in luteal tissue, but these were more than offset by increases in tissue weight. No differences in the activities in luteal tissues were detected between pregnant or pseudopregnant animals, nor between ovarian tissue adjacent to gravid or non-gravid uterine horns. The results show that significant changes occur in oestrogen metabolism in the rabbit endometrium and corpus luteum within 8 days after ovulation, and that these changes result from maternal factors expressed systemically rather than by the effects of the developing conceptus expressed locally.

Animals↗

Autotransplantation of the goat mammary gland.

The modified technique for transplanting one mammary gland of a lactating goat to the neck, with the mammary (pudic) artery and vein anastomosed to the carotid artery and jugular vein respectively, is described. This technique was successful in 6 operations and there were no significant differences between the milk yield of the transplanted and control (in situ) glands at any stage after operation. These results are compared with those from an earlier series in which no special provision was made for lymphatic drainage and in which anticoagulant therapy and treatment of the gland prior to attachment were not standardized.

Animals↗

Changes in mammary function at the onset of lactation in the goat: correlation with hormonal changes.

1. Changes in mammary function and plasma hormone concentrations during late pregnancy and the onset of lactation have been studied in conscious goats.2. Mammary blood flow, oxygen consumption and glucose uptake increased markedly and significantly between 2 days and 0.5-1 day pre-partum.3. The increase in mammary glucose uptake was relatively greater than that of oxygen consumption or blood flow.4. The concentration of citrate in mammary secretion increased; the first significant change was apparent 0.5-1 day pre-partum but the main rise occurred after this time.5. It is concluded that the marked increase in mammary glucose uptake 0.5-1 day before parturition indicates the time of onset of copious milk secretion.6. The first significant increase in the concentration of unconjugated oestrogens in arterial plasma occurred 3 days pre-partum, whereas the first significant decrease in progesterone and increase in 13,14-dihydro-15-oxoprostaglandin F(2alpha) occurred 0.5-1 day pre-partum, thus coinciding with the mammary changes; there was also a peak in prolactin concentrations at the latter time.7. The hormonal changes are discussed in relation to current concepts of the initiation of parturition and the onset of copious milk secretion in this species. It is suggested that the fall in plasma progesterone concentrations triggers milk secretion at high rates.

Animals↗

Mammary function and its control at the cessation of lactation in the goat.

1. The changes in mammary function following cessation of milking during declining lactation have been studied in conscious goats. 2. No significant changes in the rate of milk secretion, mammary blood flow or metabolism occurred in the first 24 h after cessation of milking. After then, secretory rate, mammary blood flow, oxygen consumption, glucose uptake and acetate uptake decreased markedly over the next 3 days. Up to the time of maximum udder distension on day 3, there were no major changes in milk composition. 3. It was found that the rate of milk secretion declined when the calculated pressure within the alveoli became positive. 4. After 3 days, mammary volume and intramammary pressure decreased, and the composition of milk changed slowly to resemble that of extracellular fluid, i.e. [Na+], [Cl-], [HCO3-] and pH increased while [K+], [lactose] and [citrate] decreased. During this time [lactose] and [K+] were positively correlated, and [lactose] and [Na+], and [lactose] and [Cl-] negatively correlated. 5. It is suggested that the changes in milk composition, the decreases in mammary volume and in intramammary pressure after day 3 are due to the loss of integrity of the mammary epithelium. 6. By about 7 weeks after the cessation of milking the udder volume was less than the empty udder volume before milking was stopped, indicating a loss of mammary tissue as well as the resorption of fluid. 7. When milking of an autotransplanted gland was stopped, while milking of the control gland in situ was continued, the rate of secretion in the transplant fell while that of the control did not change. 8. In goats milked normally but in which a volume of isosmotic lactose equal to the volume of milk removed at that milking was injected into the lumen of one gland at each milking, the rate of secretion of that gland, but not that of the other, decreased.

Acetates↗

Secretory activity of goat mammary glands during pregnancy and the onset of lactation.

1. The volume of the udder and the composition of the secretion have been followed in five goats through pregnancy to the onset of lactation. 2. During the middle of pregnancy udder volume was minimal and there was little or no fluid in the teats. 3. Two stages of commencing secretory activity (lactogenesis) were defined. In the first, starting up to ten weeks pre-partum, udder volume increased and the fluid in the teats changed from an extracellular-fluid-like to a milk-like composition and acquired a high concentration of immunoglobulins. Four goats accumulated several litres of a pre-colostral fluid with a high [lactose] 6-7 weeks pre-partum. 4. Comparison of the rate of increase in udder volume with previous data for the rate of increase in empty udder volume in pregnant goats showed that the rate of secretion, even in the last few days of pregnancy, was only a few per cent of the rate immediately after parturition. 5. In the last 2-3 days of pregnancy there was a three to elevenfold increase in [citrate] in the secretion; this heralded the onset of copious secretion at about the time of parturition. 6. The changes in mammary gland activities are discussed in relation to changes in plasma hormone concentrations during pregnancy.

Animals↗